WorldWideScience

Sample records for carbon disulfide

  1. 46 CFR 151.50-41 - Carbon disulfide (carbon bisulfide).

    Science.gov (United States)

    2010-10-01

    ... 46 Shipping 5 2010-10-01 2010-10-01 false Carbon disulfide (carbon bisulfide). 151.50-41 Section... CARGOES BARGES CARRYING BULK LIQUID HAZARDOUS MATERIAL CARGOES Special Requirements § 151.50-41 Carbon disulfide (carbon bisulfide). (a) All openings shall be in the top of the tank. (b) Loading lines...

  2. Brain MRI findings of carbon disulfide poisoning

    International Nuclear Information System (INIS)

    To evaluate the findings of brain MRI in patients with carbon disulfide poisoning. Ninety-one patients who had suffered carbon disulfide poisoning [male:female=87:4; age, 32-74 (mean 53.3) years] were included in this study. To determine the extent of white matter hyperintensity (Grade 0-V) and lacunar infarction, T2-weighted MR imaging of the brain was performed. T2-weighted images depicted white matter hyperintensity in 70 patients (76.9%) and lacunar infarcts in 27 (29.7%). In these patients, the prevalent findings at T2-weighted MR imaging of the brain were white matter hyperintensity and lacunar infarcts. Disturbance of the cardiovascular system by carbon disulfide might account for these results

  3. Molybdenum Disulfide Sheathed Carbon Nanotubes

    Institute of Scientific and Technical Information of China (English)

    Xu Chun SONG; Zhu De XU; Yi Fan ZHENG; Gui HAN; Bo LIU; Wei Xiang CHEN

    2004-01-01

    Single and double layered MoS2-coated multiwalled carbon nanotubes (MWCNs) were successfully prepared by pyrolyzing (NH4)2MoS4-coated multiwalled carbon nanotubes in an H2 atmosphere at 900℃. MoS2-coated MWCNs would be expected to have different tribological and mechanical properties compared to MoS2, so it may have potential applications in many fields.

  4. 40 CFR 180.467 - Carbon disulfide; tolerances for residues.

    Science.gov (United States)

    2010-07-01

    ...) PESTICIDE PROGRAMS TOLERANCES AND EXEMPTIONS FOR PESTICIDE CHEMICAL RESIDUES IN FOOD Specific Tolerances..., insecticide, and fungicide carbon disulfide, from the application of sodium tetrathiocarbonate, in or on...

  5. CuI-Catalyzed: One-Pot Synthesis of Diaryl Disulfides from Aryl Halides and Carbon Disulfide

    OpenAIRE

    Mohammad Soleiman-Beigi; Azadeh Izadi

    2013-01-01

    A new application of carbon disulfide in the presence of KF/Al2O3 is reported for the synthesis of organic symmetrical diaryl disulfides. These products were synthesized by one-pot reaction of aryl halides with the in situ generated trithiocarbonate ion in the presence of copper under air atmosphere.

  6. Peptide Bond Formation in Water Mediated by Carbon Disulfide.

    Science.gov (United States)

    Leman, Luke J; Huang, Zheng-Zheng; Ghadiri, M Reza

    2015-09-01

    Demonstrating plausible nonenzymatic polymerization mechanisms for prebiotic monomers represents a fundamental goal in prebiotic chemistry. While a great deal is now known about the potentially prebiotic synthesis of amino acids, our understanding of abiogenic polymerization processes to form polypeptides is less well developed. Here, we show that carbon disulfide (CS2), a component of volcanic emission and sulfide mineral weathering, and a widely used synthetic reagent and solvent, promotes peptide bond formation in modest yields (up to ∼20%) from α-amino acids under mild aqueous conditions. Exposure of a variety of α-amino acids to CS2 initially yields aminoacyl dithiocarbamates, which in turn generate reactive 2-thiono-5-oxazolidone intermediates, the thio analogues of N-carboxyanhydrides. Along with peptides, thiourea and thiohydantoin species are produced. Amino acid stereochemistry was preserved in the formation of peptides. Our findings reveal that CS2 could contribute to peptide bond formation, and possibly other condensation reactions, in abiogenic settings. PMID:26308392

  7. Biotechnology for removal of carbon disulfide emissions. Final report

    Energy Technology Data Exchange (ETDEWEB)

    McIntosh, M.J.

    1995-07-01

    Biological removal in a ``biofilter`` plant of carbon disulfide and hydrogen sulfide from the air effluent of a viscose plant at Teepak, Inc., is analyzed from process and economic standpoints by use of the Aspen Plus simulation program. The metabolic product from the biofilter, 3% sulfuric acid, must be transformed at the source into either a marketable or recyclable commodity (such as 95% sulfuric acid, high-quality sulfur, or high-quality gypsum) or a material with reasonable landfill costs (such as sulfur or gypsum). The simulations indicate that the total capital requirement for production of concentrated sulfuric acid is $48.9 million; for high-quality gypsum, $40.4 million; and for high-quality sulfur, $29.4 million. Production of concentrated sulfur for landfill is not economically practical. The process to neutralize the 3% acid effluent with limestone and landfill the resulting low-quality gypsum requires the lowest total investment of the processes simulated, $8.7 million, including the biofilter plant.

  8. Controlled Synthesis of Carbon Nanoparticles in a Supercritical Carbon Disulfide System

    Directory of Open Access Journals (Sweden)

    Zhengsong Lou

    2013-12-01

    Full Text Available Carbon nanoparticles with large surface areas were produced by the reduction of carbon disulfide with metallic lithium at 500 °C. The carbon nanoparticles account for about 80% of the carbon product. The carbon nanoparticles were characterized by X-ray powder diffraction, field emission scanning electron microscopy, transmission electron microscopy, high resolution transmission electron microscopy and N2 physisorption. The results showed that carbon nanoparticles predominate in the product. The influence of experimental conditions was investigated, which indicated that temperature plays a crucial role in the formation of carbon nanoparticles. The possible formation mechanism of the carbon nanoparticles was discussed. This method provides a simple and efficient route to the synthesis of carbon nanoparticles.

  9. Engineering de novo disulfide bond in bacterial α-type carbonic anhydrase for thermostable carbon sequestration

    Science.gov (United States)

    Jo, Byung Hoon; Park, Tae Yoon; Park, Hyun June; Yeon, Young Joo; Yoo, Young Je; Cha, Hyung Joon

    2016-01-01

    Exploiting carbonic anhydrase (CA), an enzyme that rapidly catalyzes carbon dioxide hydration, is an attractive biomimetic route for carbon sequestration due to its environmental compatibility and potential economic viability. However, the industrial applications of CA are strongly hampered by the unstable nature of enzymes. In this work, we introduced in silico designed, de novo disulfide bond in a bacterial α-type CA to enhance thermostability. Three variants were selected and expressed in Escherichia coli with an additional disulfide bridge. One of the variants showed great enhancement in terms of both kinetic and thermodynamic stabilities. This improvement could be attributed to the loss of conformational entropy of the unfolded state, showing increased rigidity. The variant showed an upward-shifted optimal temperature and appeared to be thermoactivated, which compensated for the lowered activity at 25 °C. Collectively, the variant constructed by the rapid and effective de novo disulfide engineering can be used as an efficient biocatalyst for carbon sequestration under high temperature conditions. PMID:27385052

  10. Engineering de novo disulfide bond in bacterial α-type carbonic anhydrase for thermostable carbon sequestration

    Science.gov (United States)

    Jo, Byung Hoon; Park, Tae Yoon; Park, Hyun June; Yeon, Young Joo; Yoo, Young Je; Cha, Hyung Joon

    2016-07-01

    Exploiting carbonic anhydrase (CA), an enzyme that rapidly catalyzes carbon dioxide hydration, is an attractive biomimetic route for carbon sequestration due to its environmental compatibility and potential economic viability. However, the industrial applications of CA are strongly hampered by the unstable nature of enzymes. In this work, we introduced in silico designed, de novo disulfide bond in a bacterial α-type CA to enhance thermostability. Three variants were selected and expressed in Escherichia coli with an additional disulfide bridge. One of the variants showed great enhancement in terms of both kinetic and thermodynamic stabilities. This improvement could be attributed to the loss of conformational entropy of the unfolded state, showing increased rigidity. The variant showed an upward-shifted optimal temperature and appeared to be thermoactivated, which compensated for the lowered activity at 25 °C. Collectively, the variant constructed by the rapid and effective de novo disulfide engineering can be used as an efficient biocatalyst for carbon sequestration under high temperature conditions.

  11. New Condensation Reaction of β-keto-δ-valerolactones, Carbon Disulfide and Alkyl Halides

    Institute of Scientific and Technical Information of China (English)

    You Ming WANG; Yu Xin LI; Su Hua WANG; Zheng Ming LI

    2004-01-01

    β-Keto-δ-valerolactones, which were obtained by reaction of acetoacetate with aldehydes or ketones, reacted with carbon disulfide, alkyl halides and a new condensation reaction was developed. The structures of the products 3 were confirmed by 1HNMR spectra and elemental analysis.

  12. Collision effects in the nonlinear Raman response of liquid carbon disulfide

    NARCIS (Netherlands)

    la Cour Jansen, T.; de Swart, M; Jensen, L.; van Duijnen, P.T.; Snijders, Jaap; Duppen, K.

    2002-01-01

    A model of the polarizability of carbon disulfide dimers was constructed, using polarizabilities from accurate time-dependent density functional theory calculations as reference. This direct reaction field model takes dipole-induced dipole effects, induced multipole effects and effects due to the ov

  13. Carbophilic versus thiophilic attack in the reaction of metallated aromates and heteroaromates with carbon disulfide

    NARCIS (Netherlands)

    Verkruijsse, H.D.; Brandsma, L.

    1987-01-01

    Copper(I) halides catalyse the formation of carbodithioates RCSSLi in the reaction of aryl- or heteroaryl-lithium reagents with carbon disulfide. Subsequent addition of methyl iodide gives the dithioesters RCSSCH3 in high yields. Appreciable amounts of the methyl sulfides RSCH3 and tars are obtained

  14. Application of Solid Phase Microextraction (SPME Sampler for Determination of Carbon Disulfide in Air

    Directory of Open Access Journals (Sweden)

    AbdulRahman Bahrami

    2015-10-01

    Full Text Available Carbon disulfide is used predominantly in the manufacture. It has affects the nervous system. In this study, the applicability of SPME as a passive sampler for determination of carbon disulphide in air was studied. Effect of sampler and environmental parameters on uptake of Carbon disulphide was studied as well. Four fibers were tested to select the best sampler for determine carbon disulfide in ambient air. A standard generation chamber was built in the laboratory and was used to test the SPME. Analysis SPME samples were carried out by a gas chromatography-Mass spectrometry and results were compared with data obtained with National Institute for Occupational Safety and Health (NIOSH method 1600. Polymethylsiloxane-carboxen (PDMS/CAR showed  the  most  effective stationary phase  material for sorbing BTEX among other materials (polyacrylate, PDMS, PDMS/divinylbenzene. Its linearity range in exposed mode was less than 10 minutes but with its retracted mode application, its linearity increased up to 8 hours. Temperature had not linear effect on uptake of pollutant in temperatures lower than 25, it has positive effect and above this range it has negative effect. Relative humidity had negative effect on mass loaded on fiber. Velocity in range of static to 0.5 m/s had no significant effect. The precision of the method was 4.18% relative standard deviation (RSD. The detection limit for carbon disulfide in the GC/MS system in SIM mode was 6.7 ng per sample. SPME is a good alternative for sampling of carbon disulfide in air. However, for the situations in high humidity values it should be used with care.

  15. Carbon tetrachloride/carbon disulfide exposures in grain fumigation. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Marano, D.E.; White, K.L.; Deer, H.; Alexander, G.

    1986-03-01

    The use of 80/20 compounds as liquid-fumigant mixtures in the grain-handling industry was considered. Worker exposures to carbon tetrachloride and carbon disulfide were measured at representative facilities throughout the industry. Work practices that may contribute to high exposure levels were cited. Approximately equal amounts of liquid fumigants and solid formulations of phosphine are used throughout the industry. As spraying is done on a sporadic basis, it does not appear likely to present a major problem such as might be encountered in continuous exposure situations. The authors conclude that grain workers do not appear to be an ideal study group for a neurotoxicity morbidity study due to the fact that the physical distance between study sites and the small number of workers at each site makes it difficult logistically. A concerted effort throughout the grain industry to educate those workers in the grain-handling portions of that industry toward the safe handling of fumigants should go far in alleviating any problems arising from exposure to fumigants.

  16. Ophthalmological and angiographic findings in workers exposed to carbon disulfide (author's transl)

    Energy Technology Data Exchange (ETDEWEB)

    Savic, S.

    1982-01-01

    Microaneurysms are important in the diagnosis of vascular changes caused by carbon disulfide. They can be diagnosed by ophtholmoscopy, angiography or angioscopy. In our opinion even a careful ophthalmoscopic investigation is sufficient for diagnosis, so that angiography is not absolutely necessary for any mass survey. The incidence of microaneurysms correlates with the duration (both daily and total) as well as with the intensity of exposure to carbon disulfide. The quantity correlates closely with the intensity of exposure. The incidence of microaneurysms is not correlated to age; however it was found to be highest in 40-50-year-old men working with staple fibers, whereas in the spinning department it occurred in 50-55-old men. Microaneurysms are found equally frequently in active workers and invalids. There was no difference between the two groups with regard to degenerative changes of the macula. However, the changes found in the eyes of men from the staple fiber department were more pronounced than in those from the spinning department.

  17. One-pot synthesis of S-alkyl dithiocarbamates via the reaction of N-tosylhydrazones, carbon disulfide and amines.

    Science.gov (United States)

    Sha, Qiang; Wei, Yun-Yang

    2013-09-14

    A new, convenient and efficient transition metal-free synthesis of S-alkyl dithiocarbamates through one-pot reaction of N-tosylhydrazones, carbon disulfide and amines is reported. Tosylhydrazones derived from various aromatic and aliphatic ketones or aldehydes were tested and gave dithiocarbamates in good to excellent yields. The tosylhydrazones can be generated in situ without isolation, which provides a simpler one-pot method to synthesize dithiocarbamates via the reaction of carbonyl compounds, carbon disulfide and amines in the presence of 4-methylbenzenesulfonohydrazide. PMID:23863979

  18. Highly Stretchable Supercapacitors Based on Aligned Carbon Nanotube/Molybdenum Disulfide Composites.

    Science.gov (United States)

    Lv, Tian; Yao, Yao; Li, Ning; Chen, Tao

    2016-08-01

    Stretchable supercapacitors that can sustain their performance under unpredictable tensile force are important elements for practical applications of various portable and wearable electronics. However, the stretchability of most reported supercapacitors was often lower than 100 % because of the limitation of the electrodes used. Herein we developed all-solid-state supercapacitors with a stretchability as high as 240 % by using aligned carbon nanotube composites with compact structure as electrodes. By combined with pseudocapacitive molybdenum disulfide nanosheets, the newly developed supercapacitor showed a specific capacitance of 13.16 F cm(-3) , and also showed excellent cycling retention (98 %) after 10 000 charge-discharge cycles. This work also presents a general and effective approach in developing high-performance electrodes for flexible and stretchable electronics. PMID:27328623

  19. Modelling a nonlinear optical switching in a standard photonic crystal fiber infiltrated with carbon disulfide

    Science.gov (United States)

    Munera, Natalia; Acuna Herrera, Rodrigo

    2016-06-01

    In this letter, a numerical analysis is developed for the propagation of ultrafast optical pulses through a standard photonic crystal fiber (PCF) consisting of two infiltrated holes using carbon disulfide (CS2). This material is a good choice since it has highly nonlinear properties, what makes it a good candidate for optical switching and broadband source at low power compared to traditional nonlinear fiber coupler. Based on supermodes theory, a set of generalized nonlinear equations is presented in order to study the propagation characteristics. It is shown in this letter that it is possible to get optical switching behavior at low power and how the dispersion, as well as, the two infiltrated holes separation influence this effect. Finally, we see that supercontinuum generation can be induced equally in both infiltrated holes despite no initial excitation at one hole.

  20. Simple one-pot synthesis of thioureas from amine, carbon disulfide and oxidants in water

    Directory of Open Access Journals (Sweden)

    Milosavljević Milutin M.

    2016-01-01

    Full Text Available The present study reports the new facile methodology for synthesis of symmetrical and asymmetrical thioureas by an one-pot reaction of amine, carbon disulfide and oxidants: hydrogen peroxide, ethylenediamine tetraacetic acid (EDTA/sodium percarbonate system or air. The structures of the synthesized compounds were confirmed by IR, 1H and 13C NMR and MS methods. Reaction mechanism has been proposed on the basis of reaction intermediate isolation and their structure determination. The synthetic benefits of the presented methods is reflected in the operational simplicity, mild reaction conditions, short reaction times, recycling of solvent, high purity and yield of products, absence of dangerous by-products and technological applicability at industrial scale. Considering commercial importance of the thioureas, it can be emphasized that implementation of the optimal synthesis of thiourea, based on presented methods, at industrial level of production would provide concurrent alternative to existing technologies in use. [Projekat Ministarstva nauke Republike Srbije, br. 172013

  1. Thio residue from thermal processing of cometary ices containing carbon disulfide and ammonia

    Science.gov (United States)

    Methikkalam, R. R. J.; Pavithraa, S.; Murali Babu, S. P.; Hill, H.; Raja Sekhar, B. N.; Pradeep, T.; Sivaraman, B.

    2016-08-01

    We have carried out experimental investigation on binary ice mixture containing carbon disulfide (CS2) and ammonia (NH3) ices formed at 10 K. Icy films were formed in various combinations to investigate the reactivity of CS2 and NH3 molecules on cometary nucleus. In the case of NH3 ices, deposition carried out at 10 K was found to contain NH3 homo-dimers that was found to reorient upon annealing to 40 K. Phase transition was found to take place as the 10 K ice was warmed to higher temperatures and the phase transition temperature was found to be 5 K higher for the mixed ice in comparison to the layered deposits. Thermal processing of the mixed deposition of CS2sbnd NH3 ice was found to leave thio residue, which could be ammonium dithiocarbamate that was even found to be present at 340 K.

  2. The flux of carbonyl sulfide and carbon disulfide between the atmosphere and a spruce forest

    Directory of Open Access Journals (Sweden)

    X. Xu

    2002-01-01

    Full Text Available Turbulent fluxes of carbonyl sulfide (COS and carbon disulfide (CS2 were measured over a spruce forest in Central Germany using the relaxed eddy accumulation (REA technique. A REA sampler was developed and validated using simultaneous measurements of CO2 fluxes by REA and by eddy correlation. REA measurements were conducted during six campaigns covering spring, summer, and fall between 1997 and 1999. Both uptake and emission of COS and CS2 by the forest were observed, with deposition occurring mainly during the sunlit period and emission mainly during the dark period. On the average, however, the forest acts as a sink for both gases. The average fluxes for COS and CS2 are  -93 ± 11.7 pmol m-2 s-1 and  -18 ± 7.6 pmol m-2 s-1, respectively. The fluxes of both gases appear to be correlated to photosynthetically active radiation and to the CO2 and chem{H_2O} fluxes, supporting the idea that the air-vegetation exchange of both gases is controlled by stomata. An uptake ratio COS/CO2 of 10 ± 1.7 pmol m mol-1 has been derived from the regression line for the correlation between the COS and CO2 fluxes. This uptake ratio, if representative for the global terrestrial net primary production, would correspond to a sink of 2.3 ± 0.5 Tg COS yr-1.

  3. Atmospheric measurements of carbonyl sulfide, dimethyl sulfide, and carbon disulfide using the electron capture sulfur detector

    Science.gov (United States)

    Johnson, James E.; Bates, Timothy S.

    1993-01-01

    Measurements of atmospheric dimethyl sulfide (DMS), carbonyl sulfide (COS), and carbon disulfide (CS2) were conducted over the Atlantic Ocean on board the NASA Electra aircraft during the Chemical Instrumentation Test and Evaluation (CITE 3) project using the electron capture sulfur detector (ECD-S). The system employed cryogenic preconcentration of air samples, gas chromatographic separation, catalytic fluorination, and electron capture detection. Samples collected for DMS analysis were scrubbed of oxidants with NaOH impregnated glass fiber filters to preconcentration. The detection limits (DL) of the system for COS, DMS, and CS2 were 5, 5, and 2 ppt, respectively. COS concentrations ranged from 404 to 603 ppt with a mean of 489 ppt for measurements over the North Atlantic Ocean (31 deg N to 41 deg N), and from 395 to 437 ppt with a mean of 419 ppt for measurements over the Tropical Atlantic Ocean (11 deg S to 2 deg N). DMS concentrations in the lower marine boundary layer, below 600-m altitude, ranged from below DL to 150 ppt from flights over the North Atlantic, and from 9 to 104 ppt over the Tropical Atlantic. CS2 concentrations ranged from below DL to 29 ppt over the North Atlantic. Almost all CS2 measurements over the Tropical Atlantic were below DL.

  4. Removal and recovery of carbon disulfide emitted by the viscose process. Final report

    Energy Technology Data Exchange (ETDEWEB)

    McIntosh, M.J.

    1992-02-01

    Teepak, Inc., which manufactures cellulose food casings by means of the viscose process, has a plant in Danville, Illinois, that emits approximately 400,000 cubic feet per minute (cfm) of water-saturated air containing approximately 100 parts per million (ppm) of carbon disulfide (CS{sub 2}). Both Teepak and the state of Illinois desire to reduce these emissions as soon as possible; however, the large air flow and very small CS{sub 2} concentration result in a difficult and costly separations problem without an obvious economically viable solution. One possibility is to incinerate the CS{sub 2}, but a more environmentally and economically acceptable alternative is to recover the CS{sub 2} for recycle to the process. The recovered CS{sub 2} would be worth about $700,000 annually to Teepak. Teepak has sponsored, with the Hazardous Waste Research and Information Center (HWRIC) of the Illinois Department of Natural Resources, a research project at Argonne National Laboratory (ANL) to evaluate current gas- purification and recovery technology and to suggest a route of development that will lead to a CS{sub 2} recovery process. The Illinois Department of Commerce and Community Affairs later provided on Illinois Challenge Grant to allow laboratory studies to supplement this effort. This report is a result of all those studies.

  5. Removal and recovery of carbon disulfide emitted by the viscose process

    Energy Technology Data Exchange (ETDEWEB)

    McIntosh, M.J.

    1992-02-01

    Teepak, Inc., which manufactures cellulose food casings by means of the viscose process, has a plant in Danville, Illinois, that emits approximately 400,000 cubic feet per minute (cfm) of water-saturated air containing approximately 100 parts per million (ppm) of carbon disulfide (CS{sub 2}). Both Teepak and the state of Illinois desire to reduce these emissions as soon as possible; however, the large air flow and very small CS{sub 2} concentration result in a difficult and costly separations problem without an obvious economically viable solution. One possibility is to incinerate the CS{sub 2}, but a more environmentally and economically acceptable alternative is to recover the CS{sub 2} for recycle to the process. The recovered CS{sub 2} would be worth about $700,000 annually to Teepak. Teepak has sponsored, with the Hazardous Waste Research and Information Center (HWRIC) of the Illinois Department of Natural Resources, a research project at Argonne National Laboratory (ANL) to evaluate current gas- purification and recovery technology and to suggest a route of development that will lead to a CS{sub 2} recovery process. The Illinois Department of Commerce and Community Affairs later provided on Illinois Challenge Grant to allow laboratory studies to supplement this effort. This report is a result of all those studies.

  6. Group separation and analysis of a carbon disulfide-soluble fraction from Shenfu coal by column chromatography

    Institute of Scientific and Technical Information of China (English)

    DING Ming-jie; WEI Xian-yong; ZONG Zhi-min; ZONG Ying; OUYANG Xiao-dong; HUANG Yao-guo; ZHOU Lei; ZHENG Yu-xuan; ZHOU Xiao; WEI Yan-bin

    2008-01-01

    A carbon disulfide-soluble fraction (CDSSF) from Shenfu coal was separated into five fractions by silica-gel column chromatography using hexane and n-hexane/ethyl acetate binary eluent. The five fractions include four clear group fractions and a nonpolar fraction. All the fractions were analyzed by GC/MS. A total of 204 compounds were detected from the original CDSSF and its further separated fractions, with 173 compounds more than those detected by studying the original CDSSF directly. The results demonstrate a clear group separation by column chromatography in coal organic components and a more accessibility to coal components compared with the solvent extraction only.

  7. Carbon doped molybdenum disulfide nanosheets stabilized on graphene for the hydrogen evolution reaction with high electrocatalytic ability

    Science.gov (United States)

    Li, Yong; Wang, Jiao; Tian, Xike; Ma, Longlong; Dai, Chu; Yang, Chao; Zhou, Zhaoxin

    2016-01-01

    Fabricating a cost effective hydrogen evolution reaction catalyst without using precious metal elements is in crucial demand for environmentally-benign energy production. In this work, the thin and edge-rich molybdenum disulfide nanosheets, with carbon doped in the interlayers and decorated on graphene, were developed by a facile solvothermal process. The as-synthesized nanohybrids exhibited high catalytic ability for the hydrogen evolution electrochemical reaction with an onset overpotential of 0.165 mV and a Tafel slope of 46 mV dec-1. Furthermore, the prepared nanohybrids also showed better durability and stability. Our work may lead to a potential method for in situ production of metal carbide-sulphur hybrid nanomaterials with promising applications for the hydrogen evolution reaction.Fabricating a cost effective hydrogen evolution reaction catalyst without using precious metal elements is in crucial demand for environmentally-benign energy production. In this work, the thin and edge-rich molybdenum disulfide nanosheets, with carbon doped in the interlayers and decorated on graphene, were developed by a facile solvothermal process. The as-synthesized nanohybrids exhibited high catalytic ability for the hydrogen evolution electrochemical reaction with an onset overpotential of 0.165 mV and a Tafel slope of 46 mV dec-1. Furthermore, the prepared nanohybrids also showed better durability and stability. Our work may lead to a potential method for in situ production of metal carbide-sulphur hybrid nanomaterials with promising applications for the hydrogen evolution reaction. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr07370g

  8. Non-enzymatic sensing of hydrogen peroxide using a glassy carbon electrode modified with a nanocomposite made from carbon nanotubes and molybdenum disulfide

    International Nuclear Information System (INIS)

    We report on a non-enzymatic electrochemical sensing strategy for ultrasensitive detection of hydrogen peroxide (H2O2) at nanomolar levels. A glassy carbon electrode (GCE) was modified with a hybrid material consisting of multiwalled carbon nanotubes (CNT) and molybdenum disulfide (MoS2). Transmission electron microscopy and Raman spectroscopy were employed to characterize the hybrid nanostructures. GCEs modified with carbon nanotubes, or nanoscaled MoS2, or with the CNT-MoS2 hybrid were investigated with respect to sensing H2O2, and this revealed that the GCE modified with the CNT-MoS2 hybrid performed best and resulted in a limit of detection as low as 5.0 nM. A repeatability and intermediate precision of 9 % was accomplished. The method was applied to determine H2O2 in spiked sterilized milk and gave satisfactory results. (author)

  9. Role of Thiobacillus thioparus in the biodegradation of carbon disulfide in a biofilter packed with a recycled organic pelletized material.

    Science.gov (United States)

    Prenafeta-Boldú, Francesc X; Rojo, Naiara; Gallastegui, Gorka; Guivernau, Miriam; Viñas, Marc; Elías, Ana

    2014-07-01

    This study reports the biodegradation of carbon disulfide (CS2) in air biofilters packed with a pelletized mixture of composted manure and sawdust. Experiments were carried out in two lab-scale (1.2 L) biofiltration units. Biofilter B was seeded with activated sludge enriched previously on CS2-degrading biomass under batch conditions, while biofilter A was left as a negative inoculation control. This inoculum was characterized by an acidic pH and sulfate accumulation, and contained Achromobacter xylosoxidans as the main putative CS2 biodegrading bacterium. Biofilter operation start-up was unsuccessfully attempted under xerophilic conditions and significant CS2 elimination was only achieved in biofilter A upon the implementation of an intermittent irrigation regime. Sustained removal efficiencies of 90-100 % at an inlet load of up to 12 g CS2 m(-3) h(-1) were reached. The CS2 removal in this biofilter was linked to the presence of the chemolithoautotrophic bacterium Thiobacillus thioparus, known among the relatively small number of species with a reported capacity of growing on CS2 as the sole energy source. DGGE molecular profiles confirmed that this microbe had become dominant in biofilter A while it was not detected in samples from biofilter B. Conventional biofilters packed with inexpensive organic materials are suited for the treatment of low-strength CS2 polluted gases (IL importance of applying culture-independent techniques for microbial community analysis as a diagnostic tool in the biofiltration of recalcitrant compounds has been highlighted. PMID:24469405

  10. Carbon Disulfide (CS2) Interference in Glucose Metabolism from Unconventional Oil and Gas Extraction and Processing Emissions.

    Science.gov (United States)

    Rich, Alisa L; Patel, Jay T; Al-Angari, Samiah S

    2016-01-01

    Carbon disulfide (CS2) has been historically associated with the manufacturing of rayon, cellophane, and carbon tetrachloride production. This study is one of the first to identify elevated atmospheric levels of CS2 above national background levels and its mechanisms to dysregulate normal glucose metabolism. Interference in glucose metabolism can indirectly cause other complications (diabetes, neurodegenerative disease, and retinopathy), which may be preventable if proper precautions are taken. Rich et al found CS2 and 12 associated sulfide compounds present in the atmosphere in residential areas where unconventional shale oil and gas extraction and processing operations were occurring. Ambient atmospheric concentrations of CS2 ranged from 0.7 parts per billion by volume (ppbv) to 103 ppbv over a continuous 24-hour monitoring period. One-hour ambient atmospheric concentrations ranged from 3.4 ppbv to 504.6 ppbv. Using the U.S. Environmental Protection Agency Urban Air Toxic Monitoring Program study as a baseline comparison for atmospheric CS2 concentrations found in this study, it was determined that CS2 atmospheric levels were consistently elevated in areas where unconventional oil and gas extraction and processing occurred. The mechanisms by which CS2 interferes in normal glucose metabolism by dysregulation of the tryptophan metabolism pathway are presented in this study. The literature review found an increased potential for alteration of normal glucose metabolism in viscose rayon occupational workers exposed to CS2. Occupational workers in the energy extraction industry exposed to CS2 and other sulfide compounds may have an increased potential for glucose metabolism interference, which has been an indicator for diabetogenic effect and other related health impacts. The recommendation of this study is for implementation of regular monitoring of blood glucose levels in CS2-exposed populations as a preventative health measure.

  11. Synthesis and structural characterization of coaxial nano tubes intercalated of molybdenum disulfide with carbon

    International Nuclear Information System (INIS)

    In this work the study of some fundamental aspects in the growth of unidimensional systems of coaxial nano tubes from the mold method is approached. This method is an inclusion technique of a precursor reagent into oxide nano porous alumina film (mold), and later applying some processes of synthesis it is gotten to obtain the wished material. The synthesized structures are identified later because they take place by means of the initial formation of nano tubes of MoS2, enclosing to carbon nano tubes by the same method, with propylene flow which generates a graphitization process that 'copy' the mold through as it flows. Binary phase MoS2 + C nano tubes were synthesized by propylene pyrolysis inside MoS2 nano tubes prepared by template assisted technique. The large coaxial nano tubes constituted of graphite sheets inserted between the MoS2 layers forming the outer part, and coaxial multi wall carbon nano tubes (MWCNT) intercalated with MoS2 inside. High resolution electron microscopy (HRTEM), electron energy loss spectroscopy (EELS), high angle annular dark field (HAADF), gatan image filter (GIF), nano beam electron diffraction patterns (NBEDP), along with molecular dynamics simulation and quantum mechanical calculations were used to characterize the samples. The one-dimensional structures exhibit diverse morphologies such as long straight and twisted nano tubes with several structural irregularities. The inter-planar spacing between MoS2 layers was found to increase from 6.3 to 7.4 A due to intercalation with carbon. Simulated HREM images revealed the presence of these twisted nano structures, with mechanical stretch into intercalate carbon between MoS2 layers. Our results open up the possibility of using MoS2 nano tubes as templates for the synthesis of new one- dimensional binary phase systems. (Author)

  12. Antibody oriented immobilization on gold using the reaction between carbon disulfide and amine groups and its application in immunosensing.

    Science.gov (United States)

    Niu, Yu; Matos, Ana I; Abrantes, Luisa M; Viana, Ana S; Jin, Gang

    2012-12-21

    Carbon disulfide (CS(2)) can spontaneously react with amine groups to form dithiocarbamates on gold surface, providing the possibility to immobilize some compounds with primary or secondary amine groups in one step. Using this principle, an immunosensor interface prepared for immunoglobulin G (IgG) sensing surface toward anti-IgG has been fabricated for the first time by simply immersing gold slides into a mixed aqueous solution of CS(2) and protein A, followed by incubation in immunoglobulin G solution. The reaction between CS(2) and protein A has been followed by UV-vis spectroscopy, whereas cyclic voltammetry has been employed in the characterization of the modified gold surface with CS(2) and protein A, both methods indicating that protein A immobilization is implemented by CS(2). Conventional ellipsometry, atomic force microscopy (AFM), as well as surface plasmon resonance (SPR) have been used to evaluate the specific binding of protein A with IgG and IgG with anti-IgG, revealing that IgG is specifically captured to form the biosensing interface, maintaining its bioactivity. Compared to direct adsorption of IgG on the gold surface, the IgG sensing surface constructed of CS(2) and protein A is far more sensitive to capture anti-IgG as its target molecule. In addition, the modified surface is proven to have good capability to inhibit nonspecific adsorption, as supported by control experiments using lysozyme and BSA. To conclude, antibody immobilization using this one-step method has potential as a simple and convenient surface modification approach for immunosensor development. PMID:23210719

  13. DNA damage and apoptosis of endometrial cells cause loss of the early embryo in mice exposed to carbon disulfide

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Bingzhen [Department of Epidemiology and Health Statistics, School of Public Health, Shandong University, Jinan (China); Shen, Chunzi [Centers for Disease Control and Prevention, Zibo (China); Yang, Liu; Li, Chunhui; Yi, Anji [Department of Epidemiology and Health Statistics, School of Public Health, Shandong University, Jinan (China); Wang, Zhiping, E-mail: zhipingw@sdu.edu.cn [Department of Epidemiology and Health Statistics, School of Public Health, Shandong University, Jinan (China)

    2013-12-01

    Carbon disulfide (CS{sub 2}) may lead to spontaneous abortion and very early pregnancy loss in women exposed in the workplace, but the mechanism remains unclear. We designed an animal model in which gestating Kunming strain mice were exposed to CS{sub 2} via i.p. on gestational day 4 (GD4). We found that the number of implanted blastocysts on GD8 was significantly reduced by each dose of 0.1 LD{sub 50} (157.85 mg/kg), 0.2 LD{sub 50} (315.7 mg/kg) and 0.4 LD{sub 50} (631.4 mg/kg). In addition, both the level of DNA damage and apoptosis rates of endometrial cells on GD4.5 were increased, showed definite dose–response relationships, and inversely related to the number of implanted blastocysts. The expressions of mRNA and protein for the Bax and caspase-3 genes in the uterine tissues on GD4.5 were up-regulated, while the expressions of mRNA and protein for the Bcl-2 gene were dose-dependently down-regulated. Our results indicated that DNA damage and apoptosis of endometrial cells were important reasons for the loss of implanted blastocysts induced by CS{sub 2}. - Highlights: • We built an animal model of CS2 exposure during blastocyst implantation. • Endometrial cells were used in the comet assay to detect DNA damage. • CS2 exposure caused DNA damage and endometrial cell apoptosis. • DNA damage and endometrial cell apoptosis were responsible for embryo loss.

  14. Biotechnology as an alternative for carbon disulfide treatment in air pollution control

    Energy Technology Data Exchange (ETDEWEB)

    Rojo, N.; Gallastegi, G.; Barona, A.; Gurtubay, L.; Elias, A. [Univ. of the Basque Country, Bilbao (Spain). Dept. of Chemical and Environmental Engineering; Gabriel Ibarra-Berastegi [Univ. of the Basque Country, Bilbao (Spain). Dept. of Nuclear Engineering and Fluid Mechanics

    2010-07-01

    The industry demand for CS{sub 2} has changed considerably over the last 2 decades and is expected to increase. This paper discussed the technical and financial feasibility of eliminating carbon disulphide (CS{sub 2}) from exhaust gases use biotechnology. The global emissions of this hazardous air pollutant are estimated to exceed 250,000 tonnes per year. However, the emission range depends on the source. The conventional technologies for treating CS{sub 2} emissions include thermal oxidation, thermo-catalytic processes or incineration. However, these technologies have drawbacks, such as high energy consumption and the generation of secondary by-products that require additional treatment. Recently, biotechnology was touted as an affordable, effective, and ecologically sound alternative to treat waste gases containing CS{sub 2}. Biological technologies based on microorganisms to biodegrade air pollutants overcome many of the disadvantages of conventional techniques and are particularly useful for the removal of relatively low concentrations of pollutants. The main properties, sources, and uses of CS{sub 2} were summarized in this paper along with alternative biotreatments for CS{sub 2}. Several applications of the technical and economical feasibility of biofilters and biotrickling filters were presented. Further research is required before their widespread industrial application. 72 refs., 3 tabs.

  15. A glassy carbon electrode modified with a nanocomposite consisting of molybdenum disulfide intercalated into self-doped polyaniline for the detection of bisphenol A

    International Nuclear Information System (INIS)

    Thin-layered molybdenum disulfide (MoS2) was intercalated, via ultrasonic exfoliation, into self-doped polyaniline (SPAN). This material, when placed on a glassy carbon electrode (GCE), exhibits excellent electrical conductivity and synergistic catalytic activity with respect to the detection of bisphenol A (BPA). The electrochemical response of the modified GCE to BPA was investigated by cyclic voltammetry and differential pulse voltammetry. Under optimal conditions, the oxidation peak current (measured best at 446 mV vs. SCE) is related to the concentration of BPA in the range from 1.0 nM to 1.0 μM, and the detection limit is 0.6 nM. (author)

  16. Diallyl disulfide enhances carbon ion beams-induced apoptotic cell death in cervical cancer cells through regulating Tap73 /ΔNp73.

    Science.gov (United States)

    Di, Cuixia; Sun, Chao; Li, Hongyan; Si, Jing; Zhang, Hong; Han, Lu; Zhao, Qiuyue; Liu, Yang; Liu, Bin; Miao, Guoying; Gan, Lu; Liu, Yuanyuan

    2015-01-01

    Diallyl disulfide (DADS), extracted from crushed garlic by steam-distillation, has been reported to provide the anticancer activity in several cancer types. However, the effect of DADS on high-LET carbon beams - induced cell death remains unknown. Therefore, we used human cervical cancer cells to elucidate the molecular effects of this diallyl sulfide. Radiotherapy remains the mainstay of treatment, especially in advanced cervical cancer and there is still space to improve the radiosensitivity to reduce radiation dosage. In this study, we found that radiation effects evoked by high-LET carbon beam was marked by inhibition of cell viability, cell cycle arrest, significant rise of apoptotic cells, regulation of transcription factor, such as p73, as well as alterations of crucial mediator of the apoptosis pathway. We further demonstrated that pretreatment of 10 µM DADS in HeLa cells exposed to radiation resulted in decrease in cell viability and increased radiosensitivity. Additionally, cells pretreated with DADS obviously inhibited the radiation-induced G2/M phase arrest, but promoted radiation-induced apoptosis. Moreover, combination DADS and the radiation exacerbated the activation of apoptosis pathways through up-regulated ration of pro-apoptotic Tap73 to anti-apoptotic ΔNp73, and its downstream proteins, such as FASLG, and APAF1. Taken together, these results suggest that DADS is a potential candidate as radio sensitive agent for cervical cancer. PMID:26505313

  17. Determination of Carbon Disulfide in Industrial Waste Gas by Capillary Gas Chromatography%毛细管气相色谱法测定工业废气中二硫化碳

    Institute of Scientific and Technical Information of China (English)

    魏伟

    2015-01-01

    A method of gas chromatography for the determination of carbon disulfide in industrial waste gas was given. After the adsorption of active carbon by carbon disulfide desorption of ethanol, separated by HP-INNOWAX capillary column, detected by hydrogen flame ionization detector. Carbon disulfide in 5. 04 ~ 50. 4 mg/L, the standard curve linear range, the correlation coefficient was 0. 9995 , when the sampling volume was 30 L, the minimum detectable concentration of carbon disulfide was 0. 009 mg/m3 , the recovery rate was 94. 9% ~104. 8%. This method was simple, good degree of separation, less interference, high sensitivity, it could meet the requirement of analysis.%建立了用气相色谱法测定工业废气中二硫化碳的方法。二硫化碳经活性炭吸附后由乙醇解吸, HP-INNOWAX毛细管色谱柱分离,经氢火焰离子化检测器检测。二硫化碳在5.04~50.4 mg/L范围内标准曲线线性良好,相关系数为0.9995,当采样体积为30 L,二硫化碳最低检出质量浓度为0.009 mg/m3,其加标回收率为94.9%~104.8%。本方法前处理简便,分离度好,干扰少,分析灵敏度高,能满足分析要求。

  18. Simultaneous determination of acrylonitrile, carbon disulfide, methyl ethyl ketone, and isobutanol leachates by purge and trap- gas chromatography-mass spectrometry

    International Nuclear Information System (INIS)

    According to the Mexican General Law of the Ecological Equilibrium and Environmental Protection, issued by the National Institute of Ecology, some chemicals such as acrylonitrile, methyl ethyl ketone, carbon disulfide, and isobutanol must be monitored in industrial residues because of their toxicity. This report describes an analytical method for the simultaneous determination of these four analytes in leachates. A purge and trap concentrator coupled to a computerized gas-chromatograph-mass selective detector was used to achieve the analysis. Quantitation measurements were based on the internal standardization method, using the area ratios of the molecular ions of the analytes and the internal standard obtained by deconvolution of the data. The scope of this method as well as the validation data is reported. The method is reliable in spite of the fact that, in some cases, the analytes or the standard coeluted with other compounds of the samples. Because the data acquisition is carried out in the scan mode it is possible to detect and identify other substances in the samples. (Author)

  19. Copper indium disulfide nanocrystals supported on carbonized chicken eggshell membranes as efficient counter electrodes for dye-sensitized solar cells

    Science.gov (United States)

    Wang, Lidan; He, Jianxin; Zhou, Mengjuan; Zhao, Shuyuan; Wang, Qian; Ding, Bin

    2016-05-01

    A domestic waste, chicken eggshell membrane (ESM), is used as a raw material to fabricate carbonized ESM loaded with chalcopyrite CuInS2 nanocrystals (denoted CESM-CuInS2) by a simple liquid impregnation and carbonization method. The CESM-CuInS2 composite possesses a natural three-dimensional macroporous network structure in which numerous CuInS2 nanocrystals with a size of about 25 nm are inlaid in carbon submicron fibers that form a microporous network. The CESM-CuInS2 composite is used as the counter electrode in a dye-sensitized solar cell (DSSC) and its photoelectric performance is tested. The DSSC with a CESM-CuInS2 counter electrode exhibits a short-circuit current density of 12.48 mA cm-2, open-circuit voltage of 0.78 V and power conversion efficiency of 5.8%; better than the corresponding values for a DSSC with a CESM counter electrode, and comparable to that of a reference DSSC with a platinum counter electrode. The favorable photoelectric performance of the CESM-CuInS2 counter electrode is attributed to its hierarchical structure, which provides a large specific surface area and numerous catalytically active sites to facilitate the oxidation of the electrolyte. This new composite material has many advantages, such as low cost and simple preparation, compared with Pt and pure CuInS2 counter electrodes.

  20. Reasoned opinion on the modification of the existing MRLs for dithiocarbamates (expressed as carbon disulfide in bulb vegetables, cucurbits and asparagus

    Directory of Open Access Journals (Sweden)

    European Food Safety Authority

    2012-07-01

    Full Text Available

    In accordance with Article 6 of Regulation (EC No 396/2005, Italy, herewith referred to as the evaluating Member State (EMS, received an application from the company BASF Italia Srl. to modify the existing MRLs for dithiocarbamate in cucurbits (edible and inedible peel, onions, shallots, garlic and asparagus, resulting from the use of metiram. In order to accommodate the intended uses, the EMS Italy proposed to raise the existing MRL in garlic from 0.5 mg/kg to 1 mg/kg; for the other crops they considered there was no need to modify the existing EU MRLs. Italy drafted an evaluation report according to Article 8 of Regulation (EC No 396/2005, which was submitted to the European Commission and forwarded to EFSA. According to EFSA the data were sufficient to derive MRL proposals of 0.6 mg/kg in garlic and 1.5 mg/kg in cucurbits (with inedible peel. For the uses on other crops a need to modify the existing EU MRLs was not identified. Adequate analytical enforcement methods are available to check the compliance of metiram residues (expressed as carbon disulfide and the relevant metabolite ethylenethiourea (ETU in the crops under consideration. EFSA concludes that, the intended use of metiram on garlic and cucurbits (with edible peel will not result in a consumer exposure exceeding the toxicological reference values and therefore is unlikely to pose a public health concern. The exposure situation for the other crops under consideration is not affected by the new uses requested.

  1. Metabolism and distribution of 14C- and 35S-labeled carbon disulfide in immature rats of different ages

    International Nuclear Information System (INIS)

    The metabolism and distribution of 14C- and 35S-CS2 was examined in 1-, 5-, 10-, 20-, 30-, and 40-day-old rats. During a 3-hr period following an ip dose of 14C-CS2, 58-83% of the dose was expired as CS2 and 4-9% was metabolized to expired CO2 depending on age. Thirty- and forty-day-old rats metabolized significantly more CS2 to CO2 and expired significantly less CS2 than 1- through 20-day-old rats. At the end of the measured expiration period, only biotransformation products of CS2, which were in part covalently bound, remained in tissues from rats of all ages. Tissue levels of 35S-CS2-derived radioactivity exceeded levels of 14C-CS2-derived radioactivity indicating that sulfur metabolites free from the carbon atom of CS2 were formed in rats as young as 1 day of age. The 35S-CS2-derived radioactivity per g of tissue and thus 35S covalently bound to tissue protein was significantly higher in 1- through 20-day-old rats than in 30- and 40-day-old rats. Twenty-four hr after dosing, up to 13 times more 35S-labeled metabolites were covalently bound in organs from 1-day-old rats than in similar organs from 40-day-old rats. The results showed that elimination of the biotransformation products of CS2, in particular the covalently binding sulfur metabolites, was prolonged in newborn rats in comparison to 40-day-old rats

  2. 生物滴滤技术处理二硫化碳机理研究%Research on the Mechanism of the Treatment of Carbon Disulfide by Bio-trickling Filter Technology

    Institute of Scientific and Technical Information of China (English)

    许杰; 陶金; 王水平; 王贤斌

    2015-01-01

    In order to study the mechanism of the treatment of carbon disulfide by biotricking filter technology , a biotricking filter during the experiment was set up.In the process of experiment , the best operating conditions of microbial treatment of carbon disulfide to be obtained was as follows: the pH was 7.5 , the spray quantity was 1.5 L/h, the concentration of air was 101.15 mg/m3 , gas residence time was 68.67 s by simulation experiment at first.After that , the optimal conditions were studied.It was investigated that the carbon disulfide in the transformation mechanism of microorganisms was to generate SO 2-4 and CO2.%为了研究生物滴滤技术处理二硫化碳的机理,我们在实验过程中搭建了一座生物滴滤塔。在进行实验的过程中,我们首先通过实验模拟获得微生物处理二硫化碳的最佳运行条件是pH为7.5,喷淋量为1.5 L/h,进气浓度为101.15 mg/m3,气体停留时间为68.67 s。随后我们就在此最佳运行条件下进行研究,并最终探讨得知二硫化碳在微生物体内的转化机理是生成SO2-4和CO2。

  3. PEDOT:PSS and glucose assisted preparation of molybdenum disulfide/single-wall carbon nanotubes counter electrode and served in dye-sensitized solar cells

    International Nuclear Information System (INIS)

    Graphical abstract: Much higher photovoltaic performance of dye-sensitized solar cell with (G-P-A) MoS2/SWCNTs counter electrode than that of Pt configuration device. - Highlights: • The (G-P-A) MoS2/SWCNTs composite were first time prepared and employed as counter electrode (CE) in Pt-free DSSC;. • The (G-P-A) MoS2/SWCNTs CE showed the low Rct of 1.46 Ω·cm2;. • The photo-electric conversion efficiency of the DSSC reached 8.14% based on the (G-P-A) MoS2/SWCNTs CE. - Abstract: A flower-like structure complexes of molybdenum disulfide/single-wall carbon nanotubes (MoS2/SWCNTs) are for the first time synthesized with glucose and poly (3, 4-ethylenedioxythiophene): poly (styrenesulfonate) (PEDOT:PSS) assisted (G-P-A) by the use of an in situ hydrothermal route, and proposed as counter electrode (CE) catalyst for Pt-free dye-sensitized solar cells (DSSCs). The DSSC assembled with the (G-P-A) MoS2/SWCNTs CE exhibits a high photo-electric conversion efficiency of 8.14% under the illumination of 100 mW·cm−2, comparable to that of the DSSC Pt-based (7.78%). Furthermore, the surface morphology of the (G-P-A) MoS2/SWCNTs complexes with flower-like structure is confirmed by using the scanning and transmission electron microscopes (SEM). The superior structural characteristic along with 3D large interconnected interstitial volume is advantageous fast mass transport for the electrolyte, and enables the (G-P-A) MoS2/SWCNTs CE to speed up the reduction of triiodide to iodide. The electrochemical performance of the sample is analyzed from cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). It indicates that the (G-P-A) MoS2/SWCNTs CE possesses excellent electrocatalytic activity in iodide/triiodide electrolyte and lower charge transfer resistance of 1.46 Ω·cm2 compared to the Pt electrode (2.44 Ω·cm2). Sum up, the (G-P-A) MoS2/SWCNTs CE can be considered as a promising alternative CE for Pt-free DSSCs

  4. Carbonyl sulfide, dimethyl sulfide and carbon disulfide in the Pearl River Delta of southern China: Impact of anthropogenic and biogenic sources

    Science.gov (United States)

    Guo, H.; Simpson, I. J.; Ding, A. J.; Wang, T.; Saunders, S. M.; Wang, T. J.; Cheng, H. R.; Barletta, B.; Meinardi, S.; Blake, D. R.; Rowland, F. S.

    2010-10-01

    Reduced sulfur compounds (RSCs) such as carbonyl sulfide (OCS), dimethyl sulfide (DMS) and carbon disulfide (CS 2) impact radiative forcing, ozone depletion, and acid rain. Although Asia is a large source of these compounds, until now a long-term study of their emission patterns has not been carried out. Here we analyze 16 months of RSC data measured at a polluted rural/coastal site in the greater Pearl River Delta (PRD) of southern China. A total of 188 canister air samples were collected from August 2001 to December 2002. The OCS and CS 2 mixing ratios within these samples were higher in autumn/winter and lower in summer due to the influence of Asian monsoon circulations. Comparatively low DMS values observed in this coastal region suggest a relatively low biological productivity during summer months. The springtime OCS levels in the study region (574 ± 40 pptv) were 25% higher than those on other East Asia coasts such Japan, whereas the springtime CS 2 and DMS mixing ratios in the PRD (47 ± 38 pptv and 22 ± 5 pptv, respectively) were 3-30 times lower than elevated values that have been measured elsewhere in East Asia (Japan and Korea) at this time of year. Poor correlations were found among the three RSCs in the whole group of 188 samples, suggesting their complex and variable sources in the region. By means of backward Lagrangian particle release simulations, air samples originating from the inner PRD, urban Hong Kong and South China Sea were identified. The mean mixing ratio of OCS in the inner PRD was significantly higher than that in Hong Kong urban air and South China Sea marine air ( p 0.05). Using a linear regression method based on correlations with the urban tracer CO, the estimated OCS emission in inner PRD (49.6 ± 4.7 Gg yr -1) was much higher than that in Hong Kong (0.32 ± 0.05 Gg yr -1), whereas the estimated CS 2 and DMS emissions in the study region accounted for a very few percentage of the total CS 2 and DMS emission in China. These

  5. Additional disulfide bonds in insulin

    DEFF Research Database (Denmark)

    Vinther, Tine N; Pettersson, Ingrid; Huus, Kasper;

    2015-01-01

    The structure of insulin, a glucose homeostasis-controlling hormone, is highly conserved in all vertebrates and stabilized by three disulfide bonds. Recently, we designed a novel insulin analogue containing a fourth disulfide bond located between positions A10-B4. The N-terminus of insulin's B......-chain is flexible and can adapt multiple conformations. We examined how well disulfide bond predictions algorithms could identify disulfide bonds in this region of insulin. In order to identify stable insulin analogues with additional disulfide bonds, which could be expressed, the Cβ cut-off distance had...... in comparison to analogues with additional disulfide bonds that were more difficult to predict. In contrast, addition of the fourth disulfide bond rendered all analogues resistant to fibrillation under stress conditions and all stable analogues bound to the insulin receptor with picomolar affinities. Thus...

  6. Multiple ways to make disulfides

    DEFF Research Database (Denmark)

    Bulleid, Neil J; Ellgaard, Lars

    2011-01-01

    Our concept of how disulfides form in proteins entering the secretory pathway has changed dramatically in recent years. The discovery of endoplasmic reticulum (ER) oxidoreductin 1 (ERO1) was followed by the demonstration that this enzyme couples oxygen reduction to de novo formation of disulfides...

  7. Synthesis and structural characterization of coaxial nano tubes intercalated of molybdenum disulfide with carbon; Sintesis y caracterizacion estructural de nanotubos coaxiales intercalados de disulfuro de molibdeno con carbono

    Energy Technology Data Exchange (ETDEWEB)

    Reza San German, C.M

    2005-07-01

    In this work the study of some fundamental aspects in the growth of unidimensional systems of coaxial nano tubes from the mold method is approached. This method is an inclusion technique of a precursor reagent into oxide nano porous alumina film (mold), and later applying some processes of synthesis it is gotten to obtain the wished material. The synthesized structures are identified later because they take place by means of the initial formation of nano tubes of MoS{sub 2}, enclosing to carbon nano tubes by the same method, with propylene flow which generates a graphitization process that 'copy' the mold through as it flows. Binary phase MoS{sub 2} + C nano tubes were synthesized by propylene pyrolysis inside MoS{sub 2} nano tubes prepared by template assisted technique. The large coaxial nano tubes constituted of graphite sheets inserted between the MoS{sub 2} layers forming the outer part, and coaxial multi wall carbon nano tubes (MWCNT) intercalated with MoS{sub 2} inside. High resolution electron microscopy (HRTEM), electron energy loss spectroscopy (EELS), high angle annular dark field (HAADF), gatan image filter (GIF), nano beam electron diffraction patterns (NBEDP), along with molecular dynamics simulation and quantum mechanical calculations were used to characterize the samples. The one-dimensional structures exhibit diverse morphologies such as long straight and twisted nano tubes with several structural irregularities. The inter-planar spacing between MoS{sub 2} layers was found to increase from 6.3 to 7.4 A due to intercalation with carbon. Simulated HREM images revealed the presence of these twisted nano structures, with mechanical stretch into intercalate carbon between MoS{sub 2} layers. Our results open up the possibility of using MoS{sub 2} nano tubes as templates for the synthesis of new one- dimensional binary phase systems. (Author)

  8. Detection rate analysis on neurological sign of workers exposed to different concentrations of carbon disulfide%接触不同浓度二硫化碳工人神经系统体征异常检出率分析

    Institute of Scientific and Technical Information of China (English)

    李奎荣; 周文慧; 谷桂珍; 周世义; 郑玉新; 余善法

    2014-01-01

    effects of exposed to different concentrations of carbon disulfide on neurological signs of workers.Methods Collection the information of concentration of carbon disulfide in the workplace or workers individuals exposed of a chemical fiber industry from 2004 to 2011,a total of 3 537 workers exposed to carbon disulfide were detected muscle strength and muscle tone,knee reflex,Achilles tendon reflex,trembling limbs,sensory function,and three chatter.Chi-square test was used for statistical analysis on abnormal neurological signs of workers.Results Eight hours time-weighted average concentration range of workers exposed to carbon disulfide in this chemical fiber industry was 0.2-41.0 mg/m3,geometric mean was 2.38 mg/m3.Concentration of carbon disulfide exposure of 1 771 workers was from 0.2 to 2.5 mg/m3 (≤ 2.5 mg/m3),642 workers was 2.6-4.8 mg/m3 (< 5.0 mg/m3),other 1 051 workers was from 5.1 to 41.0 mg/m3 (> 5.0 mg/m3) in all subjects.The different detection rates of knee reflex were 3.0% (31/1 045),3.7% (21/574),4.8% (16/331),3.3% (10/305),5.9% (11/187),6.7% (68/1 022),the different detection rates of Achilles tendon reflex were 2.2% (23/ 1 045),3.7%(21/574),2.7% (9/331),2.3% (7/305),2.1% (4/187),5.6% (57/1 022),the different detection rates of sensory dysfunction were 0.4% (4/1 045),0.5% (3/574),0.6% (2/331),0.0% (0/305),2.1% (4/187),1.7% (17/1 022) in different cumulative amount of contact groups(≤ 10.0,10.1-20.0,20.1-30.0,30.1-40.0,40.1-50.0,> 50.0 mg/m3 per year),and the differences were statistically significant(x2 =-19.53,21.27 and 15.89,all P values were < 0.01).Stratified according to age and gender,in addition to the ≤25 years group the difference of detection rate analysis on Achilles tendon reflex was statistically significant in the different concentration group(the ratio of on Achilles tendon reflex in the different groups of concentration of carbon disulfide exposure of 2.5,2.6-5.0,≥5.0 mg/m3 were 0.4% (2

  9. Polarized and depolarized Raman spectra of liquid carbon disulfide in the pressure range 0-10 kbar. I. Vibration frequencies, C-S bond length, and Fermi resonance

    Science.gov (United States)

    Ikawa, S.; Whalley, Edward

    1986-09-01

    The effect of pressure on the polarized and depolarized Raman spectra of liquid carbon disulphide, i.e., the peak frequencies, bandwidths, and relative intensities of both the allowed ν1 and 2ν2 bands and the interaction-induced ν2 and ν3 bands, have been measured at 22 °C up to 10 kbar. This paper discusses the effect of pressure on the frequencies and on the relative isotropic intensity of the ν1 and 2ν2 bands. The frequency of the ν1 band increases linearly with pressure, within the experimental uncertainty, at the rate 0.16±0.01 cm-1 kbar-1, and the frequencies of the ν2, ν3, and 2ν2 bands decrease nonlinearly. The frequency shifts are described by second-order perturbation theory with the molecular anharmonicity and the intermolecular interaction as perturbations. The leading terms of the shifts consist of the same derivative of the interaction potential, multiplied by different anharmonicity constants, and the shifts of the ν1 and 2ν2 bands suggests that the C-S bond length decreases at the rate 2×10-4 Å kbar-1. The relative isotropic intensity of the 2ν2 and ν1 bands increases with pressure at the rate 0.050 kbar-1, whereas the anisotropic 2ν2 intensity relative to the isotropic ν1 intensity is independent of pressure to the experimental precision of ˜0.005. The effect of pressure on the second derivative of the isotropic and anisotropic parts of the polarizability with respect to the bend coordinate was estimated as 1.1×10-43 C m2 V-1 kbar-1 and ˜0, respectively, from these values.

  10. Protein folding guides disulfide bond formation

    OpenAIRE

    Qin, Meng; Wang, Wei; Thirumalai, D.

    2015-01-01

    Anfinsen inferred the principles of protein folding by studying a protein containing four disulfide bonds in the native state. However, how protein folding drives disulfide bond formation is poorly understood despite the role such proteins play in variety of extracellular and intracellular functions. We developed a method to mimic the complex chemistry of disulfide bond formation in molecular simulations, which is used to decipher the mechanism of folding of bovine pancreatic trypsin inhibito...

  11. Amino Acid Patterns around Disulfide Bonds

    Directory of Open Access Journals (Sweden)

    Brett Drury

    2010-11-01

    Full Text Available Disulfide bonds provide an inexhaustible source of information on molecular evolution and biological specificity. In this work, we described the amino acid composition around disulfide bonds in a set of disulfide-rich proteins using appropriate descriptors, based on ANOVA (for all twenty natural amino acids or classes of amino acids clustered according to their chemical similarities and Scheffé (for the disulfide-rich proteins superfamilies statistics. We found that weakly hydrophilic and aromatic amino acids are quite abundant in the regions around disulfide bonds, contrary to aliphatic and hydrophobic amino acids. The density distributions (as a function of the distance to the center of the disulfide bonds for all defined entities presented an overall unimodal behavior: the densities are null at short distances, have maxima at intermediate distances and decrease for long distances. In the end, the amino acid environment around the disulfide bonds was found to be different for different superfamilies, allowing the clustering of proteins in a biologically relevant way, suggesting that this type of chemical information might be used as a tool to assess the relationship between very divergent sets of disulfide-rich proteins.

  12. Removal of Trimethylamine and Carbon Disulfide-containing Odor Gas in Trickling Biofilter%生物法处理含氮硫无机有机恶臭气体研究

    Institute of Scientific and Technical Information of China (English)

    谢志荣; 魏在山; 曾贵华; 匡婷; 邝婉文

    2011-01-01

    生物滴滤塔处理含三甲胺(TMA)和二硫化碳(CS2)的双组分含氮硫无机有机混合恶臭气体的研究结果表明,生物法能有效去除含三甲胺和二硫化碳的混合恶臭气体,三甲胺(TMA)和二硫化碳(CS2)的去除效率分别可达99.8%、93.8%,生物脱臭装置对恶臭污染物的改变具有很强的适应性,对新恶臭污染物质的进入有较好的适应性,具有较好的抗冲击负荷性,运行稳定,能适应非连续性生产的要求.污染物之间没有明显的相巧抑制作用,进气浓度的提高对三甲胺的生物降解效率影响微弱,对二硫化碳的生物降解效率影响较大.适宜空床停留时间为20.6 s,三甲胺去除几乎不受循环液pH变化的影响,二硫化碳的去除则在pH=7-8.3时较高.生物降解动力学研究表明,生物塔对三甲胺的最大去除能力优于二硫化碳,对二硫化碳的亲和力优于三甲胺.%Trickling biofilter packed with ceramsite was set up to study removal of odor containing trimethylamine (TMA) and carbon disulfide (CS2). Experimental results showed that removal efficiency of TMA and CS2 attained 99.8% and 93.8% respectively. The trickling biofilter has a good adaptability for pollutant changes, different volumetric loading and treatment odor from industrial production. The biofilter process may nm stability and adaptation to fluctuating concentrations in waste gas. The optimal empty bed residence time (EBRT) and pH of cycling water were 20.6 s and 7.0~8.3 separately. There is not obvious inhibition in TMA and CS2 removal TMA or CS2 concentration has little effect on biodegradation of TMA, but has great influence on biodegradation of CS2 The analysis of biodegradation kinetics showed that removal capacity of TMA is higher than that of CS2, showing microorganisms have more affinity to CS2 than TMA.

  13. Interchangeable modules in bacterial thiol-disulfide exchange pathways

    NARCIS (Netherlands)

    Kouwen, Thijs R. H. M.; van Dijl, Jan Maarten

    2009-01-01

    Thiol-disulfide oxidoreductases (TDORs) catalyze thiol-disulfide exchange reactions that are crucial for protein activity and stability. Specifically, they can function as thiol oxidases, disulfide reductases or disulfide isomerases. The generally established view is that particular TDORs act unidir

  14. Shedding light on disulfide bond formation

    DEFF Research Database (Denmark)

    Ostergaard, H; Henriksen, A; Hansen, F G;

    2001-01-01

    To visualize the formation of disulfide bonds in living cells, a pair of redox-active cysteines was introduced into the yellow fluorescent variant of green fluorescent protein. Formation of a disulfide bond between the two cysteines was fully reversible and resulted in a >2-fold decrease in the i...... for redox-active cysteines. In the cytoplasm of Escherichia coli, the protein was a sensitive probe for the redox changes that occur upon disruption of the thioredoxin reductive pathway....

  15. Study on mechanism of female gonad toxicity induced by carbon disulfide in rats%二硫化碳对雌性大鼠性腺毒性机制的初步研究

    Institute of Scientific and Technical Information of China (English)

    李煌元; 张文昌; 林炜; 黄寿铨; 闫平

    2001-01-01

    Objective To explore the toxic mechanism of carbon disulfide (CS2) in term of adjusting and balancing function of the hypothalamic-pituitary-ovary axis. Methods SD rats were given CS2 with dose of 400 mg/kg and 100 mg/kg for 14 days and 28 days respectively and the changes of serum follicle-stimulating-hormone (FSH), luteinizing hormone (LH), progesterone(P) and estrogen (E2) in rat estrus were measured. In addition, rat in estrus was given gonadotropic-release hormone (GnRH) to stimulate pituitary secretion. Results As exposure dose increasing and exposure time prolonging, it was showed that the time of estrus cycle in rats was delayed to certain extent and even rats' estrus seemed to be disappeared. The ratio of ovary to body weight in CS2 group was increased significantly with comparison to that of control group (P<0.05). LH was significantly increased in those two treated groups than that in control group (P<0.05). Except that serum levle of P of high dose group was significantly increased 60 minutes after injection of GnRH than that of low dose group and control group (P<0.05), there was no diference of serum hormone level among each group (P>0.05). Conclusion It concluded that CS2 might have adverse effect on female-gonad and indicated that the reservating function of pituitary in rats was not reduced in GnRH-stimulating-test, however, the possibility of GnRH super-response in rat was unable to be excluded.%目的 探讨CS2对下丘脑-垂体-卵巢轴平衡调节功能的影响机制。方法 对SD大鼠给予不同剂量CS2(每天400 mg/kg\\, 100 mg/kg)染毒不同时间(14 d、28 d)后,测定大鼠动情期血清中激素FSH、LH、P、E2含量变化及动情间期给予GnRH刺激试验。结果 随着染毒剂量的增大或染毒时间的延长,大鼠均不同程度地出现动情周期延长,甚至出现动情期消失,CS2染毒大鼠卵巢重/体重比值与对照组比较,显著增高且具有显著差异(P<0.05)

  16. On the photostability of the disulfide bond

    DEFF Research Database (Denmark)

    Stephansen, Anne Boutrup; Larsen, Martin Alex Bjørn; Klein, Liv Bærenholdt;

    2014-01-01

    Photostability is an essential property of molecular building blocks of nature. Disulfides are central in the structure determination of proteins, which is in striking contradiction to the result that the S-S bond is a photochemically labile structural entity that cleaves to form free radicals upon...... on a sub 50 fs timescale without further ado. In a cyclic motif resembling the cysteine-disulfide bond in proteins, light can perturb the S-S bond to generate short-lived diradicaloid species, but the sulfur atoms are conformationally restricted by the ring that prevents the sulfur atoms from flying apart...... the photostability of disulfide-bonds must be ascribed a cyclic structural arrangement....

  17. DISULFIND: A DISULFIDE BONDING STATE AND CYSTEINE CONNECTIVITY PREDICTION SERVER

    OpenAIRE

    Ceroni, A; Passerini, A.; Vullo,A; Frasconi, P.

    2006-01-01

    DISULFIND is a server for predicting the disulfide bonding state of cysteines and their disulfide connectivity starting from sequence alone. Optionally, disulfide connectivity can be predicted from sequence and a bonding state assignment given as input. The output is a simple visualization of the assigned bonding state (with confidence degrees) and the most likely connectivity patterns. The server is available at .

  18. Functional differences in yeast protein disulfide isomerases

    DEFF Research Database (Denmark)

    Nørgaard, P; Westphal, V; Tachibana, C;

    2001-01-01

    PDI1 is the essential gene encoding protein disulfide isomerase in yeast. The Saccharomyces cerevisiae genome, however, contains four other nonessential genes with homology to PDI1: MPD1, MPD2, EUG1, and EPS1. We have investigated the effects of simultaneous deletions of these genes. In several...

  19. Determination of Benzenes in the Drinking Water after Carbon Disulfide Extracting by Capillary Column Gas Chromatography%二硫化碳萃取毛细管柱气相色谱法测定饮用水中苯系物

    Institute of Scientific and Technical Information of China (English)

    陈军; 朱建丰; 封蓉芳

    2011-01-01

    Objective:To develop a method for determining the benzenes in drinking water by capillary column gas chromatography.Methods: Used carbon disulfide to extract the benzenes in drinking water,Gas chromatography separation and detection.Results: The recoverie%目的:建立二硫化碳萃取毛细管柱气相色谱法测定饮用水中苯系物的方法。方法:用二硫化碳萃取饮用水中的苯系物,气相色谱法进行分离检测。结果:7种苯系物的加标回收率在87.0%~111.3%之间,在0.01~1.0mg/L之间线性关系良好。相对标准偏差(RSD)在3.2%~9.3%之间。结论:该方法简单、快速、准确、灵敏,适用于饮用水中苯系物的测定。

  20. Widespread Disulfide Bonding in Proteins from Thermophilic Archaea

    Directory of Open Access Journals (Sweden)

    Julien Jorda

    2011-01-01

    Full Text Available Disulfide bonds are generally not used to stabilize proteins in the cytosolic compartments of bacteria or eukaryotic cells, owing to the chemically reducing nature of those environments. In contrast, certain thermophilic archaea use disulfide bonding as a major mechanism for protein stabilization. Here, we provide a current survey of completely sequenced genomes, applying computational methods to estimate the use of disulfide bonding across the Archaea. Microbes belonging to the Crenarchaeal branch, which are essentially all hyperthermophilic, are universally rich in disulfide bonding while lesser degrees of disulfide bonding are found among the thermophilic Euryarchaea, excluding those that are methanogenic. The results help clarify which parts of the archaeal lineage are likely to yield more examples and additional specific data on protein disulfide bonding, as increasing genomic sequencing efforts are brought to bear.

  1. Thiol-Disulfide Exchange between Glutaredoxin and Glutathione

    DEFF Research Database (Denmark)

    Iversen, Rasmus; Andersen, Peter Anders; Jensen, Kristine Steen;

    2010-01-01

    Glutaredoxins are ubiquitous thiol-disulfide oxidoreductases which catalyze the reduction of glutathione-protein mixed disulfides. Belonging to the thioredoxin family, they contain a conserved active site CXXC motif. The N-proximal active site cysteine can form a mixed disulfide with glutathione ...... has been replaced with serine. The exchange reaction between the reduced protein and oxidized glutathione leading to formation of the mixed disulfide could readily be monitored by isothermal titration calorimetry (ITC) due to the enthalpic contributions from the noncovalent interactions...... a substantial effect on the thermal stability of the protein as revealed by differential scanning calorimetry....

  2. Purification and characterization of a Bacillus megaterium disulfide reductase specific for disulfides containing pantethine 4',4"-diphosphate.

    OpenAIRE

    Swerdlow, R D; Setlow, P

    1983-01-01

    An NADH-linked disulfide reductase specific for disulfides containing pantethine 4',4"-diphosphate moieties was purified 23,000-fold to homogeneity from spores of Bacillus megaterium. The enzyme had a native molecular weight of 122,000 with two apparently identical subunits, contained one molecule of flavin adenine dinucleotide per subunit, and was inhibited by the vicinal dithiol reagent arsenite. The enzyme was active only on disulfides containing pantethine 4',4"-diphosphate moieties, incl...

  3. Disulfide bonds and glycosylation in fungal peroxidases.

    Science.gov (United States)

    Limongi, P; Kjalke, M; Vind, J; Tams, J W; Johansson, T; Welinder, K G

    1995-01-15

    Four conserved disulfide bonds and N-linked and O-linked glycans of extracellular fungal peroxidases have been identified from studies of a lignin and a manganese peroxidase from Trametes versicolor, and from Coprinus cinereus peroxidase (CIP) and recombinant C. cinereus peroxidase (rCIP) expressed in Aspergillus oryzae. The eight cysteine residues are linked 1-3, 2-7, 4-5 and 6-8, and are located differently from the four conserved disulfide bridges present in the homologous plant peroxidases. CIP and rCIP were identical in their glycosylation pattern, although the extent of glycan chain heterogeneity depended on the fermentation batch. CIP and rCIP have one N-linked glycan composed only of GlcNAc and Man at residue Asn142, and two O-linked glycans near the C-terminus. The major glycoform consists of single Man residues at Thr331 and at Ser338. T. versicolor lignin isoperoxidase TvLP10 contains a single N-linked glycan composed of (GlcNAc)2Man5 bound to Asn103, whereas (GlcNAc)2Man3 was found in T. versicolor manganese isoperoxidase TvMP2 at the same position. In addition, mass spectrometry of the C-terminal peptide of TvMP2 indicated the presence of five Man residues in O-linked glycans. No phosphate was found in these fungal peroxidases. PMID:7851395

  4. Compact conformations of human protein disulfide isomerase.

    Directory of Open Access Journals (Sweden)

    Shang Yang

    Full Text Available Protein disulfide isomerase (PDI composed of four thioredoxin-like domains a, b, b', and a', is a key enzyme catalyzing oxidative protein folding in the endoplasmic reticulum. Large scale molecular dynamics simulations starting from the crystal structures of human PDI (hPDI in the oxidized and reduced states were performed. The results indicate that hPDI adopts more compact conformations in solution than in the crystal structures, which are stabilized primarily by inter-domain interactions, including the salt bridges between domains a and b' observed for the first time. A prominent feature of the compact conformations is that the two catalytic domains a and a' can locate close enough for intra-molecular electron transfer, which was confirmed by the characterization of an intermediate with a disulfide between the two domains. Mutations, which disrupt the inter-domain interactions, lead to decreased reductase activity of hPDI. Our molecular dynamics simulations and biochemical experiments reveal the intrinsic conformational dynamics of hPDI and its biological impact.

  5. Disulfide bonds and glycosylation in fungal peroxidases.

    Science.gov (United States)

    Limongi, P; Kjalke, M; Vind, J; Tams, J W; Johansson, T; Welinder, K G

    1995-01-15

    Four conserved disulfide bonds and N-linked and O-linked glycans of extracellular fungal peroxidases have been identified from studies of a lignin and a manganese peroxidase from Trametes versicolor, and from Coprinus cinereus peroxidase (CIP) and recombinant C. cinereus peroxidase (rCIP) expressed in Aspergillus oryzae. The eight cysteine residues are linked 1-3, 2-7, 4-5 and 6-8, and are located differently from the four conserved disulfide bridges present in the homologous plant peroxidases. CIP and rCIP were identical in their glycosylation pattern, although the extent of glycan chain heterogeneity depended on the fermentation batch. CIP and rCIP have one N-linked glycan composed only of GlcNAc and Man at residue Asn142, and two O-linked glycans near the C-terminus. The major glycoform consists of single Man residues at Thr331 and at Ser338. T. versicolor lignin isoperoxidase TvLP10 contains a single N-linked glycan composed of (GlcNAc)2Man5 bound to Asn103, whereas (GlcNAc)2Man3 was found in T. versicolor manganese isoperoxidase TvMP2 at the same position. In addition, mass spectrometry of the C-terminal peptide of TvMP2 indicated the presence of five Man residues in O-linked glycans. No phosphate was found in these fungal peroxidases.

  6. Dynamic combinatorial chemistry with diselenides and disulfides in water

    DEFF Research Database (Denmark)

    Rasmussen, Brian; Sørensen, Anne; Gotfredsen, Henrik;

    2014-01-01

    Diselenide exchange is introduced as a reversible reaction in dynamic combinatorial chemistry in water. At neutral pH, diselenides are found to mix with disulfides and form dynamic combinatorial libraries of diselenides, disulfides, and selenenylsulfides. This journal is © the Partner Organisations...

  7. Intramolecular versus intermolecular disulfide bonds in prion proteins.

    Science.gov (United States)

    Welker, Ervin; Raymond, Lynne D; Scheraga, Harold A; Caughey, Byron

    2002-09-01

    Prion protein (PrP) is the major component of the partially protease-resistant aggregate that accumulates in mammals with transmissible spongiform encephalopathies. The two cysteines of the scrapie form, PrP(Sc), were found to be in their oxidized (i.e. disulfide) form (Turk, E., Teplow, D. B., Hood, L. E., and Prusiner, S. B. (1988) Eur. J. Biochem. 176, 21-30); however, uncertainty remains as to whether the disulfide bonds are intra- or intermolecular. It is demonstrated here that the monomers of PrP(Sc) are not linked by intermolecular disulfide bonds. Furthermore, evidence is provided that PrP(Sc) can induce the conversion of the oxidized, disulfide-intact form of the monomeric cellular prion protein to its protease-resistant form without the temporary breakage and subsequent re-formation of the disulfide bonds in cell-free reactions.

  8. Catalysis of protein disulfide bond isomerization in a homogeneous substrate.

    Science.gov (United States)

    Kersteen, Elizabeth A; Barrows, Seth R; Raines, Ronald T

    2005-09-13

    Protein disulfide isomerase (PDI) catalyzes the rearrangement of nonnative disulfide bonds in the endoplasmic reticulum of eukaryotic cells, a process that often limits the rate at which polypeptide chains fold into a native protein conformation. The mechanism of the reaction catalyzed by PDI is unclear. In assays involving protein substrates, the reaction appears to involve the complete reduction of some or all of its nonnative disulfide bonds followed by oxidation of the resulting dithiols. The substrates in these assays are, however, heterogeneous, which complicates mechanistic analyses. Here, we report the first analysis of disulfide bond isomerization in a homogeneous substrate. Our substrate is based on tachyplesin I, a 17-mer peptide that folds into a beta hairpin stabilized by two disulfide bonds. We describe the chemical synthesis of a variant of tachyplesin I in which its two disulfide bonds are in a nonnative state and side chains near its N and C terminus contain a fluorescence donor (tryptophan) and acceptor (N(epsilon)-dansyllysine). Fluorescence resonance energy transfer from 280 to 465 nm increases by 28-fold upon isomerization of the disulfide bonds into their native state (which has a lower E(o') = -0.313 V than does PDI). We use this continuous assay to analyze catalysis by wild-type human PDI and a variant in which the C-terminal cysteine residue within each Cys-Gly-His-Cys active site is replaced with alanine. We find that wild-type PDI catalyzes the isomerization of the substrate with kcat/K(M) = 1.7 x 10(5) M(-1) s(-1), which is the largest value yet reported for catalysis of disulfide bond isomerization. The variant, which is a poor catalyst of disulfide bond reduction and dithiol oxidation, retains virtually all of the activity of wild-type PDI in catalysis of disulfide bond isomerization. Thus, the C-terminal cysteine residues play an insignificant role in the isomerization of the disulfide bonds in nonnative tachyplesin I. We conclude

  9. Disulfide Mispairing During Proinsulin Folding in the Endoplasmic Reticulum.

    Science.gov (United States)

    Haataja, Leena; Manickam, Nandini; Soliman, Ann; Tsai, Billy; Liu, Ming; Arvan, Peter

    2016-04-01

    Proinsulin folding within the endoplasmic reticulum (ER) remains incompletely understood, but it is clear that in mutant INS gene-induced diabetes of youth (MIDY), progression of the (three) native disulfide bonds of proinsulin becomes derailed, causing insulin deficiency, β-cell ER stress, and onset of diabetes. Herein, we have undertaken a molecular dissection of proinsulin disulfide bond formation, using bioengineered proinsulins that can form only two (or even only one) of the native proinsulin disulfide bonds. In the absence of preexisting proinsulin disulfide pairing, Cys(B19)-Cys(A20) (a major determinant of ER stress response activation and proinsulin stability) preferentially initiates B-A chain disulfide bond formation, whereas Cys(B7)-Cys(A7) can initiate only under oxidizing conditions beyond that existing within the ER of β-cells. Interestingly, formation of these two "interchain" disulfide bonds demonstrates cooperativity, and together, they are sufficient to confer intracellular transport competence to proinsulin. The three most common proinsulin disulfide mispairings in the ER appear to involve Cys(A11)-Cys(A20), Cys(A7)-Cys(A20), and Cys(B19)-Cys(A11), each disrupting the critical Cys(B19)-Cys(A20) pairing. MIDY mutations inhibit Cys(B19)-Cys(A20) formation, but treatment to force oxidation of this disulfide bond improves folding and results in a small but detectable increase of proinsulin export. These data suggest possible therapeutic avenues to ameliorate ER stress and diabetes. PMID:26822090

  10. DNA Charge Transport Leading to Disulfide Bond Formation

    OpenAIRE

    Takada, Tadao; Barton, Jacqueline K.

    2005-01-01

    Here, we show that DNA-mediated charge transport (CT) can lead to the oxidation of thiols to form disulfide bonds in DNA. DNA assemblies were prepared possessing anthraquinone (AQ) as a photooxidant spatially separated on the duplex from two SH groups incorporated into the DNA backbone. Upon AQ irradiation, HPLC analysis reveals DNA ligated through a disulfide. The reaction efficiency is seen to vary in assemblies containing intervening DNA mismatches, confirming that the reaction is DNA-medi...

  11. Scalable Production of Molybdenum Disulfide Based Biosensors.

    Science.gov (United States)

    Naylor, Carl H; Kybert, Nicholas J; Schneier, Camilla; Xi, Jin; Romero, Gabriela; Saven, Jeffery G; Liu, Renyu; Johnson, A T Charlie

    2016-06-28

    We demonstrate arrays of opioid biosensors based on chemical vapor deposition grown molybdenum disulfide (MoS2) field effect transistors (FETs) coupled to a computationally redesigned, water-soluble variant of the μ-opioid receptor (MOR). By transferring dense films of monolayer MoS2 crystals onto prefabricated electrode arrays, we obtain high-quality FETs with clean surfaces that allow for reproducible protein attachment. The fabrication yield of MoS2 FETs and biosensors exceeds 95%, with an average mobility of 2.0 cm(2) V(-1) s(-1) (36 cm(2) V(-1) s(-1)) at room temperature under ambient (in vacuo). An atomic length nickel-mediated linker chemistry enables target binding events that occur very close to the MoS2 surface to maximize sensitivity. The biosensor response calibration curve for a synthetic opioid peptide known to bind to the wild-type MOR indicates binding affinity that matches values determined using traditional techniques and a limit of detection ∼3 nM (1.5 ng/mL). The combination of scalable array fabrication and rapid, precise binding readout enabled by the MoS2 transistor offers the prospect of a solid-state drug testing platform for rapid readout of the interactions between novel drugs and their intended protein targets. PMID:27227361

  12. Free-Standing Hierarchically Sandwich-Type Tungsten Disulfide Nanotubes/Graphene Anode for Lithium-Ion Batteries

    OpenAIRE

    Chen, R.; Zhao, T.; Wu, W.; Wu, F.; Li, L.; Qian, J.; Xu, R.; H. Wu; Albishri, H. M.; Al-Bogami, A. S.; El-Hady, D. A.; Lu, J; Amine, K.

    2014-01-01

    Transition metal dichalcogenides (TMD), analogue of graphene, could form various dimensionalities. Similar to carbon, one dimensional (1D) nanotube of TMD materials has wide application in hydrogen storage,Li-ion batteries and supercapacitors due to their unique structure and properties. Here we demonstrate the feasibility of tungsten disulfide nanotubes (WS2-NTs)/graphene (GS) sandwich-type architecture as anode for lithium-ion batteries for the first time. The graphene based hierarchical ar...

  13. Clinical characteristics of the patients with occupational chronic carbon disulfide poisoning in a chemical fiber factory of Nanjing%某化纤厂职业性慢性轻度二硫化碳中毒病例临床特征

    Institute of Scientific and Technical Information of China (English)

    季春萍; 朱宝立; 倪春辉; 宋海燕; 徐进; 王美林; 侯志国; 魏春龙; 董秋; 王守宇; 乔善磊

    2012-01-01

    目的 通过分析267例职业性慢性二硫化碳(CS2)中毒病例的临床特征,为修订我国CS2接触者职业性健康体检项目提供依据.方法 选择2006年4月至2010年5月某市职业病院诊断专家组依据GBZ 4-2002《职业性慢性二硫化碳中毒诊断标准》,诊断的职业性慢性轻度CS2中毒267例病例为研究对象.所有病例均来自同一化纤厂,初次诊断为CS2中毒时,由职业卫生专业人员以问卷调查采集详细病史及职业史,并进行神经系统、心血管系统检查,生化指标、神经-肌电图检测.结果 87.3%( 233/267)的病例出现腓总神经、正中神经、尺神经、胫后神经运动传导速度减慢,英中腓总神经和正中神经运动传导速度降低检出率分别为48.6%( 138/248),37.0%(155/419).中毒病例自觉症状以神经衰弱、肢体麻木、感觉异常为主,神经系统体征中跟腱反射(79.4%,212/267)、膝反射(49.8%,133/267)阳性检出率较高.心电图ST段改变(T波低平、T波倒置、ST段压低)检出率为19.1%(51/267);高血压、收缩压升高及舒张压升高检出率依次为27.3%( 73/267)、22.5%( 60/267)和21.1%( 59/267).乳酸脱氢酶、三酸甘油酯、低密度脂蛋白异常阳性检出率较高.男性病例的间接胆红素、直接胆红素、尿酸阳性检出率明显高于女性,并且尿素氮、间接胆红素阳性检出率随接毒工龄的延长而逐渐增加.结论 职业性慢性CS2中毒主要影响神经系统,对肝肾功能也有一定的影响.CS2作业人员职业性健康体检中,用神经肌-电图筛检运动神经损伤时,建议首先检查正中运动神经和腓总运动神经传导速度.%Objective To analyze the clinical characteristics of 267 cases with occupational chronic carbon disulfide (CS2) poisoning and to provide the basis for revising the items of periodical medical examination of workers occupationally exposed to CS2.Methods The subjects of present study were 267

  14. Disulfide Bridges: Bringing Together Frustrated Structure in a Bioactive Peptide.

    Science.gov (United States)

    Zhang, Yi; Schulten, Klaus; Gruebele, Martin; Bansal, Paramjit S; Wilson, David; Daly, Norelle L

    2016-04-26

    Disulfide bridges are commonly found covalent bonds that are usually believed to maintain structural stability of proteins. Here, we investigate the influence of disulfide bridges on protein dynamics through molecular dynamics simulations on the cysteine-rich trypsin inhibitor MCoTI-II with three disulfide bridges. Correlation analysis of the reduced cyclic peptide shows that two of the three disulfide distances (Cys(11)-Cys(23) and Cys(17)-Cys(29)) are anticorrelated within ∼1 μs of bridge formation or dissolution: when the peptide is in nativelike structures and one of the distances shortens to allow bond formation, the other tends to lengthen. Simulations over longer timescales, when the denatured state is less structured, do not show the anticorrelation. We propose that the native state contains structural elements that frustrate one another's folding, and that the two bridges are critical for snapping the frustrated native structure into place. In contrast, the Cys(4)-Cys(21) bridge is predicted to form together with either of the other two bridges. Indeed, experimental chromatography and nuclear magnetic resonance data show that an engineered peptide with the Cys(4)-Cys(21) bridge deleted can still fold into its near-native structure even in its noncyclic form, confirming the lesser role of the Cys(4)-Cys(21) bridge. The results highlight the importance of disulfide bridges in a small bioactive peptide to bring together frustrated structure in addition to maintaining protein structural stability. PMID:27119635

  15. Disulfide bond formation in prokaryotes: history, diversity and design.

    Science.gov (United States)

    Hatahet, Feras; Boyd, Dana; Beckwith, Jon

    2014-08-01

    The formation of structural disulfide bonds is essential for the function and stability of a great number of proteins, particularly those that are secreted. There exists a variety of dedicated cellular catalysts and pathways from archaea to humans that ensure the formation of native disulfide bonds. In this review we describe the initial discoveries of these pathways and report progress in recent years in our understanding of the diversity of these pathways in prokaryotes, including those newly discovered in some archaea. We will also discuss the various successful efforts to achieve laboratory-based evolution and design of synthetic disulfide bond formation machineries in the bacterium Escherichia coli. These latter studies have also led to new more general insights into the redox environment of the cytoplasm and bacterial cell envelope. This article is part of a Special Issue entitled: Thiol-Based Redox Processes.

  16. Synthesis, characterization and bioactivity evaluation of diallyl disulfide

    Institute of Scientific and Technical Information of China (English)

    YUAN Xin-ke; CHEN Xiao-qing; JIANG Xin-yu; NIE Ya-li

    2006-01-01

    Diallyl disulfide was synthesized by phase transfer catalyst (PTC) during microwave irradiation. The effects of different factors, such as the power of microwave irradiation, the time of microwave irradiation, PTC reagents amount and the mole ratio of reactants, on the yield of product were investigated. The structure of diallyl disulfide was characterized by infrared spectra, mass spectra and 1 H nuclear magnetic resonance. The bioactivity of diallyl disulfide was evaluated by cell viability assay on HepG2 hepatoma cells. The results show that the optimal reaction conditions are as follows: tetrabutylammonium bromide(TBAB) selected as a PTC, the mass ratio of TBAB to appears to be cytotoxic to HepG2 hepatoma cells in a dose-dependent manner.

  17. CS2吸入染毒对雄性大鼠下丘脑-垂体-性腺轴超微结构的影响及NO干预作用%Ultrastructural Changes of Hypothalamus-Pituitary-Gonad Axis after Inhalation of Carbon Disulfide in Male Rats and The Intervention of Nitric Oxide

    Institute of Scientific and Technical Information of China (English)

    季佳佳; 丁情; 周义军; 黄晓彧; 王宁; 张振; 吴翠环; 王斌; 陈国元

    2012-01-01

    [目的]探讨二硫化碳(CS2)对雄性大鼠下丘脑-垂体-性腺轴超微结构的影响及一氧化氮(NO)的干预作用.[方法]将24只雄性SD大鼠按体重随机分为4组:对照组、CS2染毒组、硝普钠(sodium nitroprusside,SNP)干预组和N-甲基-L-精氨酸(L-NMMA)干预组.除对照组外,其余3组均以1250 mg/m3 CS2进行静式吸入染毒,2h/d,5d/周,共10周,SNP和L-NMMA干预组在动物染毒结束前10d开始分别腹腔注射SNP(5mg/kg)和L-NMMA(2mg/kg),1次/d.染毒结束后,取下丘脑、垂体和睾丸组织,利用透射电子显微镜观察组织超微结构的改变.[结果] CS2染毒可造成下丘脑神经元、垂体促性腺激素细胞、生长激素细胞和睾丸支持细胞线粒体肿胀,内质网扩张,SNP对CS2引起的下丘脑、垂体、睾丸组织的损伤具有拮抗作用,而L-NMMA则进一步导致病变的发生.[结论]CS2可造成下丘脑、垂体和睾丸组织超微结构的改变,NO在此过程中发挥重要作用.%[ Objective ] To study the ultrastructural changes of hypothalamic-pituitary-gonadal axis (HPGA) in carbon disulfide (CS2) treated rats and the interventional effect of nitric oxide (NO). [ Methods ] Twenty-four Sprague-Dawley male rats were randomly divided into 4 groups as: control, CS2, sodium nitroprusside (SNP) intervention and N-methyl-L-arginine (L-NMMA) intervention groups. Three experimental groups were treated with 1250mg/m3 CS2 by inhalation for 10 weeks, 2h/d, 5d/w. Two intervention groups received intraperitoneal injection of SNP (5 mg/kg), a nitric oxide donor, and L-NMMA (2mg/kg), a nitric oxide synthase inhibitor, once a day for 10 days before the end of CS2 exposure. Then ultramicropathology technique was used to observe the ultrastructure of hypothalami, pituitaries and testes isolated from rats. [ Results ] CS2 induced swelling of hypothalamic neurons, gonadotropin cells, growth hormone cells and sertoli cells in male rats as well as widened endoplasmic reticulum. SNP

  18. Structures and related properties of helical, disulfide-stabilized peptides

    Energy Technology Data Exchange (ETDEWEB)

    Pagel, M.D. [Univ. of California, Berkeley, CA (United States). Dept. of Chemistry]|[Lawrence Berkeley Lab., CA (United States)

    1993-11-01

    The three dimensional structure of several peptides were determined by NMR spectroscopy and distance geometry calculations. Each peptide formed a predictable, rigid structure, consisting of an {alpha}-helix, a {open_quotes}scaffold{close_quotes} region which packed along one face of the helix, and two disulfide bridges which covalently connect the helix and scaffold regions. The peptide Apa-M5 was designed to constrain the M5 peptide from MLCK in a helical geometry using the apamin disulfide scaffold. This scaffold constrains the N- terminal end of the helix with two disulfide bridges and a reverse turn. Like the M5 peptide, Apa-M5 was found to bind calmodulin in a Ca{sup 2+}-dependent 1:1 stoichiometry. However, the dissociation constant of the (Apa-M5)-calmodulin complex, 107 nM, was 100-fold higher than the dissociation constant of the M5-calmodulin complex. This difference was due to a putative steric overlap between the Apa-M5 scaffold and calmodulin. The peptide Apa-Cro was designed to replace the large structural protein matrix of {lambda} Cro with the apamin disulfide scaffold. However, Apa-Cro did not bind the consensus DNA operator half-site of {lambda} Cro, probably due to a steric overlap between the Apa-Cro disulfide framework and the DNA. The amino acid sequence of the scaffold-disulfide bridge arrangement of the peptide Max was derived from the core sequence of scyllatoxin, which contains an {alpha}-helix constrained at the C-terminal end by two disulfide bridges and a two-stranded {beta}sheet scaffold. Max was shown to fold with >84% yield to form a predictable, stable structure that is similar to scyllatoxin. The folding and stability properties of Max make this scaffold and disulfide bridge arrangement an ideal candidate for the development of hybrid sequence peptides. The dynamics of a fraying C-terminal end of the helix of the peptide Apa-AlaN was determined by analysis of {sup 15}N NMR relaxation properties.

  19. Roasting of non-stoichiometric molybdenum disulfide in vacuum

    International Nuclear Information System (INIS)

    Investigation results of thermal decomposition rate and molybdenum disulfide oxidation, which was prepared by means of molybdenum and sulfur sintering depending on the temperature and composition in homogeneity region are given. It is shown that an increase in sulfur content in the melt of disulfide by 2% as compared with stoichiometric composition leads to the factor of 1.5-1.8. It is recommended that, when selecting optimal conditions for reprocessing of molybdenum sulfide raw material and when designing apparatus for roasting of concentrates different in composition in vacuum, this fact should be taken into account

  20. Preparation of tungsten disulfide motor oil and its tribological characteristics

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Through using mineral oil and synthetic oil to deploy the semisynthesis base oil, modifying the surfaces of ultrafine tungsten disulfide grains by surface chemical embellishment and adsorption embellishment to make them suspended steadily in the base oil as solid lubricating additive, and adding some function additives, the tungsten disulfide motor oil was prepared. The tribological characteristics of this kind motor oil and the well-known motor oils in our country and overseas were studied. The results show that the oil film strength of this kind of motor oil is respectively 1.06 and 1.38 times of that of shell helix ultra motor oil and great wall motor oil, and its sintering load is 1.75 and 2.33 times of that of them, and when tested under 392 N, 1 450 r/min and 30 min, the friction coefficients of friction pairs lubricated by the tungsten disulfide motor oil decrease with the increase of time, meanwhile, the diameter of worn spot is small, and the surface of worn spot is smooth, and no obvious furrows appear. The experiments indicate that the tungsten disulfide motor oil has the better antiwear, antifriction and extreme pressure properties than the well-known motor oils.

  1. Labile disulfide bonds in human placental insulin receptor

    International Nuclear Information System (INIS)

    The disulfide crosslinking pattern of human placental insulin receptor was investigated using selective reduction with tributylphosphine followed by alkylation with N-[3H]ethylmaleimide. Insulin receptor contains a single sulfhydryl group in each β subunit whose alkylation with N-[3H]ethylmaleimide inhibits receptor autophosphorylation. Alkylation is partially inhibited by ATP or the nonhydrolyzable substrate analog adenosine 5'-[β,γ-imido]triphosphate when the nucleotides are added as MN2+ complexes. Neither insulin nor 6 M guanidinium chloride renders additional sulfhydryl groups accessible to alkylation. When the receptor is reduced under drastic conditions with tributylphosphine in guanidinium chloride, 32 or the 37 sulfhydryl groups in the receptor's α subunit can be alkylated with N-[3H]ethylmaleimide. Surprisingly only three of the 10 cysteines in the β subunit become titratable under identical conditions. By using highly selective reducing conditions, the authors were able to determine quantitatively the maximum number of disulfide bridges that link the two αβ halves to form the tetrameric structures and those that couple the α to the β subunits. Liberation of two sulfhydryl groups in the α and one in the β subunit resulted in formation of αβ dimers. Free β subunit was formed when additional disulfide bond was reduced. Three models of the arrangement of the labile disulfide bonds, consistent with these findings, are proposed

  2. A kinetic study on pantetheinase inhibition by disulfides.

    Science.gov (United States)

    Pitari, G; Maurizi, G; Ascenzi, P; Ricci, G; Duprè, S

    1994-11-15

    The mammalian enzyme pantetheinase, which hydrolyzes pantetheine to pantothenic acid and cysteamine, is inhibited by many thiol reagents and activated by thiols. Two thiol groups of different reactivity and accessibility are involved in the catalytic process [Ricci, G., Nardini, M., Chiaraluce, R., Duprè, S. & Cavallini, D. (1986) Biochim. Biophys. Acta 870, 82-91]. The inhibition kinetics by some natural and synthetic disulfides [pantethine, cystamine, 5,5'-dithiobis(2-nitrobenzoic acid), 4,4'-dithiodipyridine and oxidized mercaptoethanol] has been studied by two experimental approaches, either by monitoring activity after incubation of the enzyme with the inhibitor or by determining the progress curves in the presence of substrate and inhibitor. Data reported here indicate that pantetheinase reacts irreversibly with various disulfides in a time-dependent manner with the formation of a mixed disulfide apparently preceeded by a conformational change, giving a modified E* form with new kinetic parameters. This modified form may be further competitively inhibited by disulfides interacting with the enzyme at the active site. PMID:7957261

  3. Maturation of Pseudomonas aeruginosa elastase - Formation of the disulfide bonds

    NARCIS (Netherlands)

    Braun, P; Ockhuijsen, C; Eppens, E; Koster, M; Bitter, W; Tommassen, J

    2001-01-01

    Elastase of Pseudomonas aeruginosa is synthesized as a preproenzyme. After propeptide-mediated folding in the periplasm, the proenzyme is autoproteolytically processed, prior to translocation of both the mature enzyme and the propeptide across the outer membrane. The formation of the two disulfide b

  4. Coenzyme A disulfide reductase, the primary low molecular weight disulfide reductase from Staphylococcus aureus. Purification and characterization of the native enzyme.

    Science.gov (United States)

    delCardayre, S B; Stock, K P; Newton, G L; Fahey, R C; Davies, J E

    1998-03-01

    The human pathogen Staphylococcus aureus does not utilize the glutathione thiol/disulfide redox system employed by eukaryotes and many bacteria. Instead, this organism produces CoA as its major low molecular weight thiol. We report the identification and purification of the disulfide reductase component of this thiol/disulfide redox system. Coenzyme A disulfide reductase (CoADR) catalyzes the specific reduction of CoA disulfide by NADPH. CoADR has a pH optimum of 7.5-8.0 and is a dimer of identical subunits of Mr 49,000 each. The visible absorbance spectrum is indicative of a flavoprotein with a lambdamax = 452 nm. The liberated flavin from thermally denatured enzyme was identified as flavin adenine dinucleotide. Steady-state kinetic analysis revealed that CoADR catalyzes the reduction of CoA disulfide by NADPH at pH 7.8 with a Km for NADPH of 2 muM and for CoA disulfide of 11 muM. In addition to CoA disulfide CoADR reduces 4,4'-diphosphopantethine but has no measurable ability to reduce oxidized glutathione, cystine, pantethine, or H2O2. CoADR demonstrates a sequential kinetic mechanism and employs a single active site cysteine residue that forms a stable mixed disulfide with CoA during catalysis. These data suggest that S. aureus employs a thiol/disulfide redox system based on CoA/CoA-disulfide and CoADR, an unorthodox new member of the pyridine nucleotide-disulfide reductase superfamily. PMID:9488707

  5. Disulfide Linkage Characterization of Disulfide Bond-Containing Proteins and Peptides by Reducing Electrochemistry and Mass Spectrometry

    DEFF Research Database (Denmark)

    Cramer, Christian N; Haselmann, Kim F; Olsen, Jesper V;

    2016-01-01

    Unravelling of disulfide linkage patterns is a crucial part of protein characterization, whether it is for a previously uncharacterized protein in basic research or a recombinant pharmaceutical protein. In the biopharmaceutical industry, elucidation of the cysteine connectivities is a necessity t...

  6. 胚胎植入期二硫化碳暴露对孕鼠脾脏淋巴细胞DNA损伤的影响%DNA damage of splenic lymphocytes in pregnant mice exposed to carbon disulfide in implantation phase

    Institute of Scientific and Technical Information of China (English)

    胡程霞; 张炳珍; 李春辉; 吴艳玲; 杨柳; 王志萍

    2013-01-01

    目的 研究胚胎植入期二硫化碳(CS2)暴露对孕鼠脾脏淋巴细胞DNA损伤的影响,从免疫损伤角度探讨CS2致胚胎植入障碍的机制.方法 分别建立不同暴露剂量和不同暴露时间2个动物模型:模型1为不同CS2暴露剂量,即在小鼠受孕第4天(GD4),分别给予4组小鼠一次性腹腔注射低(0.1LD50,157.8 mg/kg)、中(0.2LD50,315.7 mg/kg)、高(0.4LD50,631.4 mg/kg)剂量的CS2和橄榄油对照组;模型2为不同CS2暴露时间,即4组小鼠分别在GD3、GD4、GD5和GD6一次性腹腔注射CS2(0.4LD50,631.4 mg/kg),各组设平行对照.两模型对照组均注射等体积橄榄油.实验终点,制备胚胎植入期小鼠脾脏淋巴细胞单细胞悬液,用台盼蓝法检测细胞活性,并进行碱性单细胞凝胶电泳(SCGE)试验,检测孕鼠脾脏淋巴细胞DNA损伤状况.结果 (1)中、高剂量组的DNA损伤程度与对照组相比差异均有统计学意义(P<0.01);反映DNA损伤程度的各指标与暴露剂量之间均存在回归关系,且差异有统计学意义(P<0.01).(2)GD3、GD4、GD5和GD6暴露组的DNA损伤程度与对照组比较,差异均有统计学意义(P<0.01),且GD4暴露组DNA损伤程度最明显.结论 胚胎植入期CS2暴露可导致孕鼠脾脏淋巴细胞的DNA损伤,并且呈明显的剂量反应关系;GD4可能是CS2暴露致孕鼠脾脏淋巴细胞DNA损伤的敏感时间点.%Objective To investigate the DNA damage of splenic lymphocytes in pregnant mice exposed to carbon disulfide (CS2) in the implantation phase and to explore the mechanism of abnormal implantation induced by CS2 from the perspective of immune injury.Methods Mice were exposed to CS2 at different doses or at different time points in the implantation phase to establish model 1 and model 2.For model 1,mice were assigned to four groups to receive a single intraperitoneal injection of low-dose CS2 (0.1 LD50,157.8mg/kg),middle-dose CS2 (0.2 LD50,315.7 mg/kg),and high-dose CS2 (0.4 LD50,631.4 mg

  7. 国家最大容许浓度内二硫化碳暴露对血压和心电图的影响%Effects of carbon disulfide on blood pressure and electrocardiogram for workers exposed to levels below the national maximum allowable concentration

    Institute of Scientific and Technical Information of China (English)

    陈嘉; 李拥军; 杨文萍

    2009-01-01

    Objective To study the effects of carbon disulfide exposure within the national maximum allowable concentration(MAC) on blood pressure and electrocardiogram, and associations with selected factors. Methods Workers in a chemical fiber factory were divided into two groups based on the type of work: a high exposure group (HEG) of 821 individuals and a low exposure group (LEG) of 259. The CS_2 concentration at workplace was controlled under the national MAC. A set of 250 randomly selected people taking routine phys-ical check-ups in the same period and hospital constituted the control group. The systolic blood pressure (SBP) and diastolic hlood pressure (DBP) were measured on the arm, and the pulse pressure (PP) and mean arterial blood pressure (MABP) were calculated based on SBP and DBP. The blood pressure data, along with the results of the routine 12-lead electrocardiography taken at rest and records on gender, age, years of work, type of work, and concentrations of triglycerol, cholesterol, and glucose in blood, were compiled for analyses. Risk factors upon CS_2 exposure for the increase of blood pressure and occurrence of electrocardiogram abnor-malities were identified and rationalized. Results Significant difference (P<0.01) in the average values of SBP, DBP, MABP, and the corresponding abnormality incident rates was found between HEG and LEG, and between HEG and the control group. For both HEG and LEG, the incident rate of DBP abnormality(high DBP) is nearly two times as high as that of SBP. Type of work is the largest risk factor in both the high SBP and high DBP subgroups, with odds ratios (OR) of 2.086 and 2.331 respectively, and high CS_2 exposure presents more than double the risk than low exposure. On the incident rate of ECG abnormalities, beth exposure groups are significantly different (P<0.01) to the control group. High SBP in LEG and high DBP in HEG were found to be significant risk factors (OR = 3.531 and 1.638 respectively), while blood glucose

  8. 胚胎植入期二硫化碳染毒对孕鼠血清雌激素及子宫雌激素受体-α表达的影响%Effects of carbon disulfide exposure during peri-implantation on estrogen receptor-α expression in uterus and serum level of estrogen in pregnant mice

    Institute of Scientific and Technical Information of China (English)

    张炳珍; 吴艳玲; 代炳芹; 李春辉; 杨柳; 王志萍

    2013-01-01

    Objective To evaluate the effects of carbon disulfide (CS2) exposure during peri-implantation on the estrogen receptor-α (ER-α) expression in the uterus and serum level of estradiol (E2) in pregnant mice,and to explore the mechanism of embryotoxicity of CS2.Methods Healthy female mice were exposed to a single dose of CS2 (631.4 mg/kg) or olive oil (solvent control) on gestational day (GD)3,GD4,GDS,or GD6.At different time points after exposure,the serum E2 levels of the pregnant mice were measured by enzymelinked immunosorbent assay,and the expression levels of ER-α in the uterus were measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot.Results Compared with the control group,the GD3,GD4,GD5,and GD6 exposure groups showed significantly decreased serum E2 levels on day 7 of gestation (P<0.05); the GD3 and GD5 exposure groups showed significantly decreased serum E2 levels on day 6 of gestation (P<0.05).The expression level of ER-α in the GD 4 exposure group was 23.6% lower than that in the control group on day 5 of gestation,and the expression level of ER-o in the GD 5 exposure group was 72.9% lower than that in the control group on day 6 of gestation (P<0.05); the GD 3 and GD 6 exposure groups showed lower expression levels of ER-α than the control group at any time point,but no significant difference was found (P>0.05).Conclusion CS2 exposure during peri-implantation can reduce the ER-α expression in the uterus and the serum level of E2 in pregnant mice,which may be one of the mechanisms of embryotoxicity of CS2.%目的 观察胚胎植入期二硫化碳(CS2)暴露对孕鼠血清雌二醇(E2)含量和子宫组织雌激素受体α(ER-α)表达水平的影响,探讨CS2致胚胎毒性的作用机制.方法 健康雌性小鼠分别在受孕后第3天(GD3)、第4天(GD4)、第5天(GD5)、第6天(GD6)暴露于单一剂量(631.4 mg/kg)的CS2或橄榄油(溶剂对照),单次暴露之后在不同的观察终点结束实

  9. Tibial dyschondroplasia in growing chickens experimentally intoxicated with tetramethylthiuram disulfide.

    Science.gov (United States)

    Vargas, M I; Lamas, J M; Alvarenga, V

    1983-07-01

    Graded levels of tetramethylthiuram disulfide (0, 30, 60, 120, and 240 ppm) were incorporated into a broiler starter ration fed to chickens from one day old to 8 weeks of age. Clinical signs of leg abnormalities were observed as early as 5 days after the beginning of the trial. After the 3rd week, the joints were shown to present lesions, especially in the femorotibial articulation, comparable to the ones found in perosis. Tibiotarsus and other organs from the birds were examined for pathological changes at weekly intervals. Histologically, the tibiotarsus has shown an osteochondrodystrophy identical to that of tibial dyschondroplasia (TD), although it varied according to the level of tetramethylthiuram disulfide. The histopathology of the thyroid gland of the chickens involved in the present experiment will be reported in a separate manuscript. PMID:6622364

  10. Scorpion venom peptides with no disulfide bridges: a review.

    Science.gov (United States)

    Almaaytah, Ammar; Albalas, Qosay

    2014-01-01

    Scorpion venoms are rich sources of biologically active peptides that are classified into disulfide-bridged peptides (DBPs) and non-disulfide-bridged peptides (NDBPs). DBPs are the main scorpion venom components responsible for the neurotoxic effects observed during scorpion envenomation as they usually target membrane bound ion channels of excitable and non-excitable cells. Several hundred DBPs have been identified and functionally characterized in the past two decades. The NDBPs represent a novel group of molecules that have gained great interest only recently due to their high diversity both in their primary structures and bioactivities. This review provides an overview of scorpion NDBPs focusing on their therapeutic applications, modes of discovery, mechanisms of NDBPs genetic diversity and structural properties. It also provides a simple classification for NDBPs that could be adopted and applied to other NDBPs identified in future studies. PMID:24184590

  11. A degradable polydopamine coating based on disulfide-exchange reaction

    Science.gov (United States)

    Hong, Daewha; Lee, Hojae; Kim, Beom Jin; Park, Taegyun; Choi, Ji Yu; Park, Matthew; Lee, Juno; Cho, Hyeoncheol; Hong, Seok-Pyo; Yang, Sung Ho; Jung, Sun Ho; Ko, Sung-Bo; Choi, Insung S.

    2015-11-01

    Although the programmed degradation of biocompatible films finds applications in various fields including biomedical and bionanotechnological areas, coating methods have generally been limited to be substrate-specific, not applicable to any kinds of substrates. In this paper, we report a dopamine derivative, which allows for both universal coating of various substrates and stimuli-responsive film degradation, inspired by mussel-adhesive proteins. Two dopamine moieties are linked together by the disulfide bond, the cleavage of which enables the programmed film degradation. Mechanistic analysis of the degradable films indicates that the initial cleavage of the disulfide linkage causes rapid uptake of water molecules, hydrating the films, which leads to rapid degradation. Our substrate-independent coating of degradable films provides an advanced tool for drug delivery systems, tissue engineering, and anti-fouling strategies.Although the programmed degradation of biocompatible films finds applications in various fields including biomedical and bionanotechnological areas, coating methods have generally been limited to be substrate-specific, not applicable to any kinds of substrates. In this paper, we report a dopamine derivative, which allows for both universal coating of various substrates and stimuli-responsive film degradation, inspired by mussel-adhesive proteins. Two dopamine moieties are linked together by the disulfide bond, the cleavage of which enables the programmed film degradation. Mechanistic analysis of the degradable films indicates that the initial cleavage of the disulfide linkage causes rapid uptake of water molecules, hydrating the films, which leads to rapid degradation. Our substrate-independent coating of degradable films provides an advanced tool for drug delivery systems, tissue engineering, and anti-fouling strategies. Electronic supplementary information (ESI) available: Synthesis, characterization, and other additional details. See DOI: 10

  12. Graphene and molybdenum disulfide hybrids: synthesis and applications

    OpenAIRE

    Nanjundan Ashok Kumar; Mushtaq Ahmad Dar; Rukhsana Gul; Jong-Beom Baek

    2015-01-01

    Graphene and related inorganic two-dimensional (2D) nanomaterials are an exceptional class of compounds with exotic properties that are technologically intriguing. While graphene itself is chemically inert and a gapless semimetal, its isostructural analog, molybdenum disulfide (MOS2) is chemically versatile with band gaps, thereby finding significant use in a myriad of applications. Although these 2D nanomaterials individually possess tremendous authority for various applications, the combina...

  13. Labile disulfide bonds in human placental insulin receptor.

    OpenAIRE

    Finn, F. M.; Ridge, K D; HOFMANN, K

    1990-01-01

    The disulfide crosslinking pattern of human placental insulin receptor was investigated using selective reduction with tributylphosphine followed by alkylation with N-[3H]ethylmaleimide. Insulin receptor contains a single sulfhydryl group in each beta subunit whose alkylation with N-[3H]ethylmaleimide inhibits receptor autophosphorylation. Alkylation is partially inhibited by ATP or the nonhydrolyzable substrate analog adenosine 5'-[beta,gamma-imido]triphosphate when the nucleotides are added...

  14. Proinsulin Disulfide Maturation and Misfolding in the Endoplasmic Reticulum*

    OpenAIRE

    Liu, Ming; Li, Yulin; Cavener, Douglas; Arvan, Peter

    2005-01-01

    Upon nonreducing Tris-Tricine-urea-SDS-PAGE, newly synthesized proinsulin from pancreatic islets of normal rodents forms a band fast mobility representing the native disulfide isomer, which is efficiently secreted. In addition at least two slower migrating “isomer 1 and 2” bands are recovered, not discernible under reducing conditions, which represent minor species that exhibit less efficient secretion. Although rats and mice have two proinsulin genes, three distinct migrating species are als...

  15. Protein disulfide isomerase isomerizes non-native disulfide bonds in human proinsulin independent of its peptide-binding activity

    OpenAIRE

    Winter, Jeannette; Gleiter, Stefan; Klappa, Peter; Lilie, Hauke

    2011-01-01

    Protein disulfide isomerase (PDI) supports proinsulin folding as chaperone and isomerase. Here, we focus on how the two PDI functions influence individual steps in the complex folding process of proinsulin. We generated a PDI mutant (PDI-aba′c) where the b′ domain was partially deleted, thus abolishing peptide binding but maintaining a PDI-like redox potential. PDI-aba′c catalyzes the folding of human proinsulin by increasing the rate of formation and the final yield of native proinsulin. Imp...

  16. Identification of thioredoxin target disulfides in proteins released from barley aleurone layers

    DEFF Research Database (Denmark)

    Hägglund, Per; Bunkenborg, J.; Yang, Fen;

    2010-01-01

    Thioredoxins are ubiquitous disulfide reductases involved in a wide range of cellular processes including DNA synthesis, oxidative stress response and apoptosis. In cereal seeds thioredoxins are proposed to facilitate the germination process by reducing disulfide bonds in storage proteins and oth...... targets in the starchy endosperm. Here we have applied a thiol-specific labeling approach to identify specific disulfide targets of barley thioredoxin in proteins released from barley aleurone layers incubated in buffer containing gibberellic acid....

  17. Contribution of Disulfide Bridges to the Thermostability of a Type A Feruloyl Esterase from Aspergillus usamii

    OpenAIRE

    Xin Yin; Die Hu; Jian-Fang Li; Yao He; Tian-Di Zhu; Min-Chen Wu

    2015-01-01

    The contribution of disulfide bridges to the thermostability of a type A feruloyl esterase (AuFaeA) from Aspergillus usamii E001 was studied by introducing an extra disulfide bridge or eliminating a native one from the enzyme. MODIP and DbD, two computational tools that can predict the possible disulfide bridges in proteins for thermostability improvement, and molecular dynamics (MD) simulations were used to design the extra disulfide bridge. One residue pair A126-N152 was chosen, and the res...

  18. Insulin analog with additional disulfide bond has increased stability and preserved activity

    DEFF Research Database (Denmark)

    Vinther, Tine N.; Norrman, Mathias; Ribel, Ulla;

    2013-01-01

    Insulin is a key hormone controlling glucose homeostasis. All known vertebrate insulin analogs have a classical structure with three 100% conserved disulfide bonds that are essential for structural stability and thus the function of insulin. It might be hypothesized that an additional disulfide...... bond may enhance insulin structural stability which would be highly desirable in a pharmaceutical use. To address this hypothesis, we designed insulin with an additional interchain disulfide bond in positions A10/B4 based on Cα-Cα distances, solvent exposure, and side-chain orientation in human insulin...... of an insulin analog featuring a fourth disulfide bond with increased structural stability and retained function....

  19. The influence of gold(i) on the mechanism of thiolate, disulfide exchange.

    Science.gov (United States)

    Garusinghe, Gamage S P; Bessey, S Max; Bruce, Alice E; Bruce, Mitchell R M

    2016-07-28

    The mechanism of gold(i)-thiolate, disulfide exchange was investigated by using initial-rate kinetic studies, 2D ((1)H-(1)H) ROESY NMR spectroscopy, and electrochemical/chemical techniques. The rate law for exchange is overall second order, first order in gold(i)-thiolate and disulfide. 2D NMR experiments show evidence of association between gold(i)-thiolate and disulfide. Electrochemical/chemical investigations do not show evidence of free thiolate and are consistent with a mechanism involving formation of a [Au-S, S-S], four-centered metallacycle intermediate during gold(i)-thiolate, disulfide exchange. PMID:27353236

  20. Domain architecture of protein-disulfide isomerase facilitates its dual role as an oxidase and an isomerase in Ero1p-mediated disulfide formation

    DEFF Research Database (Denmark)

    Kulp, M. S.; Frickel, E. M.; Ellgaard, Lars;

    2006-01-01

    reduction/rearrangement of non-native disulfides is poorly understood. We analyzed the role of individual PDI domains in disulfide bond formation in a reaction driven by their natural oxidant, Ero1p. We found that Ero1p oxidizes the isolated PDI catalytic thioredoxin domains, A and A' at the same rate...... catalytic (A) domain. The specific order of thioredoxin domains in PDI is important in establishing the asymmetry in the rate of oxidation of the two active sites thus allowing A and A', two thioredoxin domains that are similar in sequence and structure, to serve opposing functional roles as a disulfide...

  1. PVC DISULFIDE AS CATHODE MATERIALS FOR SECONDARY LITHIUM BATTERIES

    Institute of Scientific and Technical Information of China (English)

    Guo-xiang Xu; Lu Qi; Bi-tao Yu; Lei Wen

    2006-01-01

    PVC disulfide (2SPVC) was synthesized by solution crosslink and its molecular structure was confirmed by the particle size of d0.5 = 11.3 μm. With SEM (Scanning Electron Microscope) experiment the surface morphology and obvious S-S redox reaction in charge-discharge process. When 2SPVC was used as cathode material for secondary lithium mixture of o-xylene (oxy), diglyme (DG) and dimethoxymethane (DME) at 30℃, the first discharge capacity of 2SPVC is very promising cathode candidate for rechargeable lithium batteries.

  2. Synthesis and Structure of a Novel Disulfide-Containing Aniline

    Institute of Scientific and Technical Information of China (English)

    DENG,Shi-Ren; WU,Lei; WANG,Hao; ZHOU,Bin; LI,Zao-Ying

    2004-01-01

    @@ A novel disulfide-containing aniline, 8-dihydro-1H,4H-2,3,6,7-tetrathia-anthracen-9-ylamine (5) was synthesized.The single-crystal X-ray analysis of 4 indicates that the molecular has a non-planar structure, with its four sulfur atoms out of the plane of benzene ting. The designed molecular has the advantage of high theoretic specific capacity and reversibility,[1,2] when it is to be polymerized and used as the cathode material of the secondary lithium batteries.

  3. Inclusion of a cobalt tetraazamacrocycle into layered molybdenum disulfide

    International Nuclear Information System (INIS)

    We report on the intercalation of meso-5, 5, 7, 12,12,14-hexamethyl-1,4,8,11-tetraazacyclotetradecane-1,8-diacetate complexed with Co (III) into layered molybdenum disulfide. This is achieved by using the exfoliation and re-stacking properties of LiMoS2. The cobalt complex dissolves readily in a water/acetone solvent mixture. Addition of this solution to an aqueous suspension of single MoS2 layers results in the entrapment of the cobalt macrocycle between the sheets of MoS2. The resulting intercalate was characterized by powder X-ray diffraction and thermogravimetric analysis.

  4. Wet chemical thinning of molybdenum disulfide down to its monolayer

    Directory of Open Access Journals (Sweden)

    Kiran Kumar Amara

    2014-09-01

    Full Text Available We report on the preparation of mono- and bi-layer molybdenum disulfide (MoS2 from a bulk crystal by facile wet chemical etching. We show that concentrated nitric acid (HNO3 effectively etches thin MoS2 crystals from their edges via formation of MoO3. Interestingly, etching of thin crystals on a substrate leaves behind unreacted mono- and bilayer sheets. The flakes obtained by chemical etching exhibit electronic quality comparable to that of mechanically exfoliated counterparts. Our findings indicate that the self-limiting chemical etching is a promising top-down route to preparing atomically thin crystals from bulk layer compounds.

  5. Legionella pneumophila utilizes a Single Player Disulfide-Bond Oxidoreductase System to Manage Disulfide Bond Formation and Isomerization

    Science.gov (United States)

    Kpadeh, Zegbeh Z.; Day, Shandra R.; Mills, Brandy W.; Hoffman, Paul S.

    2015-01-01

    Legionella pneumophila uses a single homodimeric disulfide bond (DSB) oxidoreductase DsbA2 to catalyze extracytoplasmic protein folding and to correct DSB errors through protein-disulfide isomerase (PDI) activity. In Escherichia coli, these functions are separated to avoid futile cycling. In L. pneumophila, DsbA2 is maintained as a mixture of disulfides (S-S) and free thiols (SH), but when expressed in E. coli, only the SH form is observed. We provide evidence to suggest that structural differences in DsbB oxidases (LpDsbB1 and LpDsbB2) and DsbD reductases (LpDsbD1 and LpDsbD2) (compared to E. coli) permit bifunctional activities without creating a futile cycle. LpdsbB1 and LpdsbB2 partially complemented an EcdsbB mutant while neither LpdsbD1 nor LpdsbD2 complemented an EcdsbD mutant unless DsbA2 was also expressed. When the dsb genes of E. coli were replaced with those of L. pneumophila, motility was restored and DsbA2 was present as a mixture of redox forms. A dominant-negative approach to interfere with DsbA2 function in L. pneumophila determined that DSB oxidase activity was necessary for intracellular multiplication and assembly/function of the Dot/Icm Type IVb secretion system. Our studies show that a single-player system may escape the futile cycle trap by limiting transfer of reducing equivalents from LpDsbDs to DsbA2. PMID:25534767

  6. Diversity of the Epsilonproteobacteria Dsb (disulfide bond systems

    Directory of Open Access Journals (Sweden)

    Katarzyna Marta Bocian-Ostrzycka

    2015-06-01

    Full Text Available The bacterial proteins of the Dsb family – important components of the posttranslational protein modification system – catalyze the formation of disulfide bridges, a process that is crucial for protein structure stabilization and activity. Dsb systems play an essential role in the assembly of many virulence factors. Recent rapid advances in global analysis of bacteria have thrown light on the enormous diversity among bacterial Dsb systems. While the Escherichia coli disulfide bond-forming system is quite well understood, the mechanisms of action of Dsb systems in other bacteria, including members of class Epsilonproteobacteria that contain pathogenic and non-pathogenic bacteria colonizing extremely diverse ecological niches, are poorly characterized. Here we present a review of current knowledge on Epsilonproteobacteria Dsb systems. We have focused on the Dsb systems of Campylobacter spp. and Helicobacter spp. because our knowledge about Dsb proteins of Wolinella and Arcobacter spp. is still scarce and comes mainly from bioinformatic studies. Helicobacter pylori is a common human pathogen that colonizes the gastric epithelium of humans with severe consequences. Campylobacter spp. is a leading cause of zoonotic enteric bacterial infections in most developed and developing nations. We focus on various aspects of the diversity of the Dsb systems and their influence on pathogenicity, particularly because Dsb proteins are considered as potential targets for a new class of anti-virulence drugs to treat human infections by Campylobacter or Helicobacter spp.

  7. Regulation of interleukin-4 signaling by extracellular reduction of intramolecular disulfides

    International Nuclear Information System (INIS)

    Interleukin-4 (IL-4) contains three structurally important intramolecular disulfides that are required for the bioactivity of the cytokine. We show that the cell surface of HeLa cells and endotoxin-activated monocytes can reduce IL-4 intramolecular disulfides in the extracellular space and inhibit binding of IL-4 to the IL-4Rα receptor. IL-4 disulfides were in vitro reduced by thioredoxin 1 (Trx1) and protein disulfide isomerase (PDI). Reduction of IL-4 disulfides by the cell surface of HeLa cells was inhibited by auranofin, an inhibitor of thioredoxin reductase that is an electron donor to both Trx1 and PDI. Both Trx1 and PDI have been shown to be located at the cell surface and our data suggests that these enzymes are involved in catalyzing reduction of IL-4 disulfides. The pro-drug N-acetylcysteine (NAC) that promotes T-helper type 1 responses was also shown to mediate the reduction of IL-4 disulfides. Our data provides evidence for a novel redox dependent pathway for regulation of cytokine activity by extracellular reduction of intramolecular disulfides at the cell surface by members of the thioredoxin enzyme family.

  8. Preparation of Vesicles and Nanoparticles of Amphiphilic Cyclodextrins Containing Labile Disulfide Bonds

    NARCIS (Netherlands)

    Nolan, Darren; Darcy, Raphael; Ravoo, Bart Jan

    2003-01-01

    Amphiphilic cyclodextrin derivatives were prepared in which a disulfide bond connects the hydrophobic substituents to the macrocycle. These compounds were obtained by 1,3-dicyclohexylcarbodiimide-mediated coupling reactions of heptakis(6-amino-6-deoxy)-B-cyclodextrins and disulfide-containing carbox

  9. Specific heat of rhenium disulfide in 360-510 K temperature range

    International Nuclear Information System (INIS)

    Isobaric specific heat of rhenium disulfide was measured by the method of differential scanning calorimetry in the range of 360-510 K. Thermodynamic functions of rhenium disulfide were calculated using literature data on standard entropy of the compound for the range of 296.15-600 K. 3 refs., 1 fig., 2 tabs

  10. The Chemistry of Alk-1-yn-1-yl DisulfidesA Review

    DEFF Research Database (Denmark)

    Senning, Alexander Erich Eugen

    2009-01-01

    The preparation and the properties of the elusive alk-1-yn-1-yl disulfides are reviewed, including the most recent quantum chemical findings with regard to their reactivity.......The preparation and the properties of the elusive alk-1-yn-1-yl disulfides are reviewed, including the most recent quantum chemical findings with regard to their reactivity....

  11. Rapid expansion of the protein disulfide isomerase gene family facilitates the folding of venom peptides

    DEFF Research Database (Denmark)

    Safavi-Hemami, Helena; Li, Qing; Jackson, Ronneshia L;

    2016-01-01

    Formation of correct disulfide bonds in the endoplasmic reticulum is a crucial step for folding proteins destined for secretion. Protein disulfide isomerases (PDIs) play a central role in this process. We report a previously unidentified, hypervariable family of PDIs that represents the most...

  12. Increasing the reactivity of an artificial dithiol-disulfide pair through modification of the electrostatic milieu

    DEFF Research Database (Denmark)

    Hansen, Rosa E; Østergaard, Henrik; Winther, Jakob R

    2005-01-01

    The thiol-disulfide exchange reaction plays a central role in the formation of disulfide bonds in newly synthesized proteins and is involved in many aspects of cellular metabolism. Because the thiolate form of the cysteine residue is the key reactive species, its electrostatic milieu is thought...... to play a key role in determining the rates of thiol disulfide exchange reactions. While modest reactivity effects have previously been seen in peptide model studies, here, we show that introduction of positive charges can have dramatic effects on disulfide bond formation on a structurally restricted....... Introduction of positively charged amino acids in the proximity of the two cysteines resulted in an up to 13-fold increase in reactivity toward glutathione disulfide. Determination of the individual pK(a) values of the cysteines showed that the observed increase in reactivity was caused by a decrease in the p...

  13. DBCP: a web server for disulfide bonding connectivity pattern prediction without the prior knowledge of the bonding state of cysteines

    OpenAIRE

    Lin, Hsuan-Hung; Tseng, Lin-Yu

    2010-01-01

    The proper prediction of the location of disulfide bridges is efficient in helping to solve the protein folding problem. Most of the previous works on the prediction of disulfide connectivity pattern use the prior knowledge of the bonding state of cysteines. The DBCP web server provides prediction of disulfide bonding connectivity pattern without the prior knowledge of the bonding state of cysteines. The method used in this server improves the accuracy of disulfide connectivity pattern predic...

  14. Cell-free synthesis system suitable for disulfide-containing proteins

    Energy Technology Data Exchange (ETDEWEB)

    Matsuda, Takayoshi [NMR Pipeline Methodology Team, RIKEN Systems and Structural Biology Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045 (Japan); Cell-Free Technology Application Laboratory, RIKEN Innovation Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045 (Japan); Watanabe, Satoru [NMR Pipeline Methodology Team, RIKEN Systems and Structural Biology Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045 (Japan); Kigawa, Takanori, E-mail: kigawa@riken.jp [NMR Pipeline Methodology Team, RIKEN Systems and Structural Biology Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045 (Japan); Cell-Free Technology Application Laboratory, RIKEN Innovation Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045 (Japan); Department of Computational Intelligence and Systems Science, Interdisciplinary Graduate School of Science and Engineering, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8502 (Japan)

    2013-02-08

    Highlights: ► Cell-free synthesis system suitable for disulfide-containing proteins is proposed. ► Disulfide bond formation was facilitated by the use of glutathione buffer. ► DsbC catalyzed the efficient shuffling of incorrectly formed disulfide bonds. ► Milligram quantities of functional {sup 15}N-labeled BPTI and lysozyme C were obtained. ► Synthesized proteins were both catalytically functional and properly folded. -- Abstract: Many important therapeutic targets are secreted proteins with multiple disulfide bonds, such as antibodies, cytokines, hormones, and proteases. The preparation of these proteins for structural and functional analyses using cell-based expression systems still suffers from several issues, such as inefficiency, low yield, and difficulty in stable-isotope labeling. The cell-free (or in vitro) protein synthesis system has become a useful protein production method. The openness of the cell-free system allows direct control of the reaction environment to promote protein folding, making it well suited for the synthesis of disulfide-containing proteins. In this study, we developed the Escherichia coli (E. coli) cell lysate-based cell-free synthesis system for disulfide-containing proteins, which can produce sufficient amounts of functional proteins for NMR analyses. Disulfide bond formation was facilitated by the use of glutathione buffer. In addition, disulfide isomerase, DsbC, catalyzed the efficient shuffling of incorrectly formed disulfide bonds during the protein synthesis reaction. We successfully synthesized milligram quantities of functional {sup 15}N-labeled higher eukaryotic proteins, bovine pancreatic trypsin inhibitor (BPTI) and human lysozyme C (LYZ). The NMR spectra and functional analyses indicated that the synthesized proteins are both catalytically functional and properly folded. Thus, the cell-free system is useful for the synthesis of disulfide-containing proteins for structural and functional analyses.

  15. Action of Mercaptan and Disulfide in Hydrogen Atom Exchange Reactions

    International Nuclear Information System (INIS)

    Free- radical, photochemical, and high-energy radiation-induced reactions may be catalysed or inhibited by rapid hydrogen atom exchange reactions of mercaptans and disulfides. The radical-induced, light-initiated, and benzophenone-sensitized decarbonylations of aldehydes are catalysed by mercaptans. The chain-propagating hydrogen transfer reaction, R' + RCH = O -> RH + RC = O , is made more rapid by a similar sequence of hydrogen atom transfers involving the sulfur compound: R' + C6H5CH2SH -> RH + C6H5CH2S'; C6H5CH2S + RCH = 0 -> C6H5CH2SH + RC = 0. The photoreduction of benzophenone in 2-propanol leads to benzpinacol by a non-chain reaction via the radicals (C6H5)2C-OH and (CH3)2COH. The reaction is retarded and inhibited by mercaptan and disulfide, which reconvert the radicals to the starting materials by rapid hydrogen transfer reactions and are themselves regenerated in their alternate valence states, each molecule of sulfur compound negating the chemical consequences of many quanta: (C6H5)2C-OH + AS' -> (C6H5)2C = O + ASH; (CH3)2C-OH + ASH -> (CH3)2C = 0 + AS'. Proof of the mechanism is found in: equilibration of initially present mercaptan or disulfide during inhibition; in racemization of optically active alcohol during inhibition; in deuterium exchange during inhibition. Similar inhibition is seen when only one intermediate radical is formed, as in the benzophenone- benzhydrol and acetophenone-α-methyl-benzyl alcohol systems. Inhibition by sulfur compounds, by the same mechanism, is found in the 60Co γ-ray induced conversion of benzophenone to benzpinacol; naphthalene has no protecting effect on benzophenone in the 60Co system, while quenching the photochemical reaction. The protection by sulfur compounds of solutes against radiation damage thus results from hydrogen atom transfer reactions. The photoreduction of benzophenone in an ether is also inhibited by the sulfur compounds, by hydrogen atom transfer reactions. A mechanism exists in this system

  16. Graphene and molybdenum disulfide hybrids: synthesis and applications

    Directory of Open Access Journals (Sweden)

    Nanjundan Ashok Kumar

    2015-06-01

    Full Text Available Graphene and related inorganic two-dimensional (2D nanomaterials are an exceptional class of compounds with exotic properties that are technologically intriguing. While graphene itself is chemically inert and a gapless semimetal, its isostructural analog, molybdenum disulfide (MOS2 is chemically versatile with band gaps, thereby finding significant use in a myriad of applications. Although these 2D nanomaterials individually possess tremendous authority for various applications, the combination of these materials in the recent past has created a new paradigm in emerging applications. Here, we summarize the current state-of-the-art and progress over the past three years on the development of hybrids of these layered materials. We highlight their pivotal role in electrochemical energy storage, sensing, hydrogen generation by photochemical water splitting and electronic device applications such as field-effect transistors. Perspectives on the challenges and opportunities for the exploration of these 2D layered hybrid materials are put forward.

  17. Simple Formation of Nanostructured Molybdenum Disulfide Thin Films by Electrodeposition

    Directory of Open Access Journals (Sweden)

    S. K. Ghosh

    2013-01-01

    Full Text Available Nanostructured molybdenum disulfide thin films were deposited on various substrates by direct current (DC electrolysis form aqueous electrolyte containing molybdate and sulfide ions. Post deposition annealing at higher temperatures in the range 450–700°C transformed the as-deposited amorphous films to nanocrystalline structure. High temperature X-ray diffraction studies clearly recorded the crystal structure transformations associated with grain growth with increase in annealing temperature. Surface morphology investigations revealed featureless structure in case of as-deposited surface; upon annealing it converts into a surface with protruding nanotubes, nanorods, or dumbbell shape nanofeatures. UV-visible and FTIR spectra confirmed about the presence of Mo-S bonding in the deposited films. Transmission electron microscopic examination showed that the annealed MoS2 films consist of nanoballs, nanoribbons, and multiple wall nanotubes.

  18. Dynamic Combinatorial Chemistry with Diselenides, Disulfides, Imines and Metal Coordination

    DEFF Research Database (Denmark)

    Sørensen, Anne

    The design and preparation of strong and selective artificial receptors, especially biomi-metic receptors that function in aqueous solution, has proved truly challenging. In this thesis it will be described how the strengths of dynamic combinatorial chemistry can be used to great advantage...... in this field. The aim of this project has therefore been to develop new ways of using dynamic combinatorial libraries for molecular recognition in aqueous media. The focus has been on using what has been learned from the well-established di-sulfide exchange chemistry to incorporate a new reaction into dynamic...... combinatorial chemistry, namely the reversible diselenide exchange reaction. The first part of the thesis describes the development of a thermally induced OAr → SeAr migration reaction. Here, it was proven possible to rearrange a variety of substituted O-aryl selenocarbamates into the corresponding Se...

  19. Tuning thermal conductivity in molybdenum disulfide by electrochemical intercalation

    Science.gov (United States)

    Zhu, Gaohua; Liu, Jun; Zheng, Qiye; Zhang, Ruigang; Li, Dongyao; Banerjee, Debasish; Cahill, David G.

    2016-01-01

    Thermal conductivity of two-dimensional (2D) materials is of interest for energy storage, nanoelectronics and optoelectronics. Here, we report that the thermal conductivity of molybdenum disulfide can be modified by electrochemical intercalation. We observe distinct behaviour for thin films with vertically aligned basal planes and natural bulk crystals with basal planes aligned parallel to the surface. The thermal conductivity is measured as a function of the degree of lithiation, using time-domain thermoreflectance. The change of thermal conductivity correlates with the lithiation-induced structural and compositional disorder. We further show that the ratio of the in-plane to through-plane thermal conductivity of bulk crystal is enhanced by the disorder. These results suggest that stacking disorder and mixture of phases is an effective mechanism to modify the anisotropic thermal conductivity of 2D materials. PMID:27767030

  20. Novel alkyl substituted polyanilines/molybdenum disulfide nanocomposites

    International Nuclear Information System (INIS)

    Polyaniline (PANI), poly(N-methyl aniline) (PMA), poly(ethyl aniline) (PEA) and poly(propyl aniline) (PPA) were synthesized in their salt form, and then characterized by FT-IR spectroscopy and charge transport measurements. The solubility of the polymers was tested in a variety of solvents and N-methylformamide (NMF) was found to be the best solvent. While polyaniline gave a colloidal suspension in NMF, the solubility of the polymer increased with increasing length of the alkyl group, resulting in a concomitant decrease in electrical conductivity. The solubility of the polymers was exploited and their intercalation was performed in molybdenum disulfide by using the exfoliating/restacking property of LiMoS2. Powder X-ray diffraction showed that genuine intercalation compounds were formed. The resulting nanocomposites were also characterized by thermogravimetric analysis (TGA)

  1. Selective and efficient electrochemical biosensing of ultrathin molybdenum disulfide sheets

    Science.gov (United States)

    Narayanan, Tharangattu N.; Vusa, Chiranjeevi S. R.; Alwarappan, Subbiah

    2014-08-01

    Atomically thin molybdenum disulfide (MoS2) sheets were synthesized and isolated via solvent-assisted chemical exfoliation. The charge-dependent electrochemical activities of these MoS2 sheets were studied using positively charged hexamine ruthenium (III) chloride and negatively charged ferricyanide/ferrocyanide redox probes. Ultrathin MoS2 sheet-based electrodes were employed for the electrochemical detection of an important neurotransmitter, namely dopamine (DA), in the presence of ascorbic acid (AA). MoS2 electrodes were identified as being capable of distinguishing the coexistence of the DA and the AA with an excellent stability. Moreover, the enzymatic detection of the glucose was studied by immobilizing glucose oxidase on the MoS2. This study opens enzymatic and non-enzymatic electrochemical biosensing applications of atomic MoS2 sheets, which will supplement their established electronic applications.

  2. Cyclic disulfide C8 iminoporfiromycin: nucleophilic activation of a porfiromycin.

    Science.gov (United States)

    Lee, Sang Hyup; Kohn, Harold

    2004-04-01

    The clinical success of mitomycin C (1) and its associated toxicities and resistance have led to efforts to prepare semisynthetic analogues (i.e., KW-2149 (3), BMS-181174 (4)) that have improved pharmacological profiles. In this study, we report the preparation and evaluation of the novel 7-N-(1'-amino-4',5'-dithian-2'-yl)porfiromycin C(8) cyclized imine (6) and its reference compound, 7-N-(1'-aminocyclohex-2'-yl)porfiromycin C(8) cyclized imine (13). Porfiromycin 6 contains a disulfide unit that, upon cleavage, may provide thiol(s) that affect drug reactivity. We demonstrated that phosphines dramatically accelerated 6 activation and solvolysis in methanolic solutions ("pH 7.4") compared with 13. Porfiromycins 6 and 13 efficiently cross-linked EcoRI-linearized pBR322 DNA upon addition of Et3P. We found enhanced levels of interstrand cross-link (ISC) adducts for 6 and 13 compared with porfiromycin (7) and that 6 was more efficient than 13. The large Et3P-mediated rate enhancements for the solvolysis of 6 compared with 13 and a N(7)-substituted analogue of 1, and the increased levels of ISC adducts for 6 compared with 13 and 7 are attributed to a nucleophile-assisted disulfide cleavage process that permits porfiromycin activation and nucleophile (MeOH, DNA) adduction. The in vitro antiproliferative activities of 6 and 13 using the A549 tumor cell line (lung adenocarcinoma) were determined under aerobic and hypoxic conditions and then compared with 7. Both 6 and 13 were more cytotoxic than 7, with 13 being more potent than 6. The C(8) iminoporfiromycins 6 and 13 displayed anticancer profiles similar to 3. PMID:15053618

  3. Human β-Defensin 4 with Non-Native Disulfide Bridges Exhibit Antimicrobial Activity

    Science.gov (United States)

    Sharma, Himanshu; Nagaraj, Ramakrishnan

    2015-01-01

    Human defensins play multiple roles in innate immunity including direct antimicrobial killing and immunomodulatory activity. They have three disulfide bridges which contribute to the stability of three anti-parallel β-strands. The exact role of disulfide bridges and canonical β-structure in the antimicrobial action is not yet fully understood. In this study, we have explored the antimicrobial activity of human β-defensin 4 (HBD4) analogs that differ in the number and connectivity of disulfide bridges. The cysteine framework was similar to the disulfide bridges present in μ-conotoxins, an unrelated class of peptide toxins. All the analogs possessed enhanced antimicrobial potency as compared to native HBD4. Among the analogs, the single disulfide bridged peptide showed maximum potency. However, there were no marked differences in the secondary structure of the analogs. Subtle variations were observed in the localization and membrane interaction of the analogs with bacteria and Candida albicans, suggesting a role for disulfide bridges in modulating their antimicrobial action. All analogs accumulated in the cytosol where they can bind to anionic molecules such as nucleic acids which would affect several cellular processes leading to cell death. Our study strongly suggests that native disulfide bridges or the canonical β-strands in defensins have not evolved for maximal activity but they play important roles in determining their antimicrobial potency. PMID:25785690

  4. Disulfide Trapping for Modeling and Structure Determination of Receptor:Chemokine Complexes

    Science.gov (United States)

    Kufareva, Irina; Gustavsson, Martin; Holden, Lauren G.; Qin, Ling; Zheng, Yi; Handel, Tracy M.

    2016-01-01

    Despite the recent breakthrough advances in GPCR crystallography, structure determination of protein-protein complexes involving chemokine receptors and their endogenous chemokine ligands remains challenging. Here we describe disulfide trapping, a methodology for generating irreversible covalent binary protein complexes from unbound protein partners by introducing two cysteine residues, one per interaction partner, at selected positions within their interaction interface. Disulfide trapping can serve at least two distinct purposes: (i) stabilization of the complex to assist structural studies, and/or (ii) determination of pairwise residue proximities to guide molecular modeling. Methods for characterization of disulfide-trapped complexes are described and evaluated in terms of throughput, sensitivity, and specificity towards the most energetically favorable cross-links. Due to abundance of native disulfide bonds at receptor:chemokine interfaces, disulfide trapping of their complexes can be associated with intramolecular disulfide shuffling and result in misfolding of the component proteins; because of this, evidence from several experiments is typically needed to firmly establish a positive disulfide crosslink. An optimal pipeline that maximizes throughput and minimizes time and costs by early triage of unsuccessful candidate constructs is proposed. PMID:26921956

  5. Determination of disulfide bridges of two spider toxins: hainantoxin-III and hainantoxin-IV

    Directory of Open Access Journals (Sweden)

    W Wang

    2009-01-01

    Full Text Available Peptide toxins are usually highly bridged proteins with multipairs of intrachain disulfide bonds. Analysis of disulfide connectivity is an important facet of protein structure determination. In this paper, we successfully assigned the disulfide linkage of two novel peptide toxins, called HNTX-III and HNTX-IV, isolated from the venom of Ornithoctonus hainana spider. Both peptides are useful inhibitors of TTX-sensitive voltage-gated sodium channels and are composed of six cysteine residues that form three disulfide bonds, respectively. Firstly, the peptides were partially reduced by tris(2-carboxyethyl-phosphine (TCEP in 0.1 M citrate buffer containing 6 M guanidine-HCl at 40° C for ten minutes. Subsequently, the partially reduced intermediates containing free thiols were separated by reversed-phase high-performance liquid chromatography (RP-HPLC and alkylated by rapid carboxamidomethylation. Then, the disulfide bonds of the intermediates were analyzed by Edman degradation. By using the strategy above, disulfide linkages of HNTX-III and HNTX-IV were determined as I-IV, II-V and III-VI pattern. In addition, this study also showed that this method may have a great potential for determining the disulfide bonds of spider peptide toxins.

  6. Highly efficient supercapacitor electrode with two-dimensional tungsten disulfide and reduced graphene oxide hybrid nanosheets

    Science.gov (United States)

    Tu, Chao-Chi; Lin, Lu-Yin; Xiao, Bing-Chang; Chen, Yu-Shiang

    2016-07-01

    Two-dimensional (2D) nanostructures with their high surface area and large in-plane conductivity have been regarded as promising materials for supercapacitors (SCs). Tungsten disulfide (WS2) is highly suitable for charge accumulation with its abundant active sites in the interspacing between the 2D structures and the intraspacing of each atomic layer, as well as on the tungsten centers with the charges generated by the Faradaic reactions. This study proposes the preparation of well-constructed WS2/reduced graphene oxide (RGO) nanosheets using a simple molten salt process as the electroactive material for SCs, which presents a high specific capacitance (CF) of 2508.07 F g-1 at the scan rate of 1 mV s-1, because of the synergic effect of WS2 with its large charge-accumulating sites on the 2D planes and RGO with its highly enhanced conductivity and improved connections in the WS2 networks. The excellent cycling stability of 98.6% retention after 5000 cycles charge/discharge process and the Coulombic efficiency close to 100% for the entire measurement are also achieved for the WS2/RGO-based SC electrode. The results suggest the potential for the combination of the 2D metal sulfide and carbon materials as the charge storage material to solve the energy problems and attain a sustainable society.

  7. Heterologous expression of five disulfide-bonded insecticidal spider peptides.

    Science.gov (United States)

    Estrada, Georgina; Silva, Anita O; Villegas, Elba; Ortiz, Ernesto; Beirão, Paulo S L; Corzo, Gerardo

    2016-09-01

    The genes of the five disulfide-bonded peptide toxins 1 and 2 (named Oxytoxins or Oxotoxins) from the spider Oxyopes lineatus were cloned into the expression vector pQE30 containing a 6His-tag and a Factor Xa proteolytic cleavage region. These two recombinant vectors were transfected into Escherichia coli BL21 cells and expressed under induction with isopropyl thiogalactoside (IPTG). The product of each gene was named HisrOxyTx1 or HisrOxyTx2, and the protein expression was ca 14 and 6 mg/L of culture medium, respectively. Either recombinant toxin HisrOxyTx1 or HisrOxyTx2 were found exclusively in inclusion bodies, which were solubilized using a chaotropic agent, and then, purified using affinity chromatography and reverse-phase HPLC (RP-HPLC). The HisrOxyTx1 and HisrOxyTx2 products, obtained from the affinity chromatographic step, showed several peptide fractions having the same molecular mass of 9913.1 and 8030.1 Da, respectively, indicating that both HisrOxyTx1 and HisrOxyTx2 were oxidized forming several distinct disulfide bridge arrangements. The isoforms of both HisrOxyTx1 and HisrOxyTx2 after DTT reduction eluted from the column as a single protein component of 9923 and 8040 Da, respectively. In vitro folding of either HisrOxyTx1 or HisrOxyTx2 yielded single oxidized components, which were cleaved independently by the proteolytic enzyme Factor Xa to give the recombinant peptides rOxyTx1 and rOxyTx2. The experimental molecular masses of rOxyTx1 and rOxyTx2 were 8059.0 and 6176.4 Da, respectively, which agree with their expected theoretical masses. The recombinant peptides rOxyTx1 and rOxyTx2 showed lower but comparable toxicity to the native toxins when injected into lepidopteran larvae; furthermore, rOxyTx1 was able to inhibit calcium ion currents on dorsal unpaired median (DUM) neurons from Periplaneta americana. PMID:27263806

  8. Rapid expansion of the protein disulfide isomerase gene family facilitates the folding of venom peptides.

    Science.gov (United States)

    Safavi-Hemami, Helena; Li, Qing; Jackson, Ronneshia L; Song, Albert S; Boomsma, Wouter; Bandyopadhyay, Pradip K; Gruber, Christian W; Purcell, Anthony W; Yandell, Mark; Olivera, Baldomero M; Ellgaard, Lars

    2016-03-22

    Formation of correct disulfide bonds in the endoplasmic reticulum is a crucial step for folding proteins destined for secretion. Protein disulfide isomerases (PDIs) play a central role in this process. We report a previously unidentified, hypervariable family of PDIs that represents the most diverse gene family of oxidoreductases described in a single genus to date. These enzymes are highly expressed specifically in the venom glands of predatory cone snails, animals that synthesize a remarkably diverse set of cysteine-rich peptide toxins (conotoxins). Enzymes in this PDI family, termed conotoxin-specific PDIs, significantly and differentially accelerate the kinetics of disulfide-bond formation of several conotoxins. Our results are consistent with a unique biological scenario associated with protein folding: The diversification of a family of foldases can be correlated with the rapid evolution of an unprecedented diversity of disulfide-rich structural domains expressed by venomous marine snails in the superfamily Conoidea. PMID:26957604

  9. Simultaneous Disulfide and Boronic Acid Ester Exchange in Dynamic Combinatorial Libraries

    DEFF Research Database (Denmark)

    Diemer, Sanna L.; Kristensen, Morten; Rasmussen, Brian;

    2015-01-01

    that operate simultaneously or two reversible reactions that operate independently. Both these scenarios have advantages and disadvantages. In this contribution, we show how disulfide exchange and boronic ester transesterification can function simultaneous in dynamic combinatorial libraries under appropriate...

  10. In-source photocatalytic reduction of disulfide bonds during laser desorption ionization

    OpenAIRE

    Qiao, L.; Bi, H.; Busnel, J M; B. Liu; Girault, H. H.

    2008-01-01

    A photosensitive plate based on sintered TiO2 nanoparticles has been developed to carry out in-source photo-induced reductions for cleavage of disulfide bridges using glucose as a hole scavenger during laser desorption ionization.

  11. Shedding light on disulfide bond formation: engineering a redox switch in green fluorescent protein

    DEFF Research Database (Denmark)

    Østergaard, H.; Henriksen, A.; Hansen, Flemming G.;

    2001-01-01

    To visualize the formation of disulfide bonds in living cells, a pair of redox-active cysteines was introduced into the yellow fluorescent variant of green fluorescent protein. Formation of a disulfide bond between the two cysteines was fully reversible and resulted in a >2-fold decrease in the i......To visualize the formation of disulfide bonds in living cells, a pair of redox-active cysteines was introduced into the yellow fluorescent variant of green fluorescent protein. Formation of a disulfide bond between the two cysteines was fully reversible and resulted in a >2-fold decrease...... as a structural reorganization of residues in the immediate chromophore environment. By combining this information with spectroscopic data, we propose a detailed mechanism accounting for the observed redox state-dependent fluorescence. The redox potential of the cysteine couple was found to be within...

  12. Conformational landscape and pathway of disulfide bond reduction of human alpha defensin

    NARCIS (Netherlands)

    Snijder, Joost; Van De Waterbeemd, Michiel; Glover, Matthew S.; Shi, Liuqing; Clemmer, David E.; Heck, Albert J R

    2015-01-01

    Human alpha defensins are a class of antimicrobial peptides with additional antiviral activity. Such antimicrobial peptides constitute a major part of mammalian innate immunity. Alpha defensins contain six cysteines, which form three well defined disulfide bridges under oxidizing conditions. Residue

  13. Electrochemistry-Assisted Top-Down Characterization of Disulfide-Containing Proteins

    OpenAIRE

    Zhang, Yun; Cui, Weidong; Zhang, Hao; Dewald, Howard D.; Chen, Hao

    2012-01-01

    Covalent disulfide bond linkage in a protein represents an important challenge for mass spectrometry (MS)-based top-down protein structure analysis as it reduces the backbone cleavage efficiency for MS/MS dissociation. This study presents a strategy for solving this critical issue via integrating electrochemistry (EC) online with top-down MS approach. In this approach, proteins undergo electrolytic reduction in an electrochemical cell to break disulfide bonds and then online ionized into gase...

  14. Vicinal Dithiol-Disulfide Distribution in the Escherichia coli Mannitol Specific Carrier Enzyme IImtl

    OpenAIRE

    Roossien, F.F.; Robillard, G. T.

    1984-01-01

    Escherichia coli mannitol specific EII in membrane vesicles can be inhibited by the action of the oxidizable substrate-reduced phenazine methosulfate (PMS) in a manner similar to E. coli enzyme IIGlc. The fact that reduced PMS and various oxidizing agents protect the enzyme from inactivation by the sulfhydryl reagents N-ethylmaleimide and bromopyruvate suggests that the active form possesses a dithiol which can be protected by conversion to a disulfide. The sulfhydryl-disulfide distribution h...

  15. Regulation of pyruvate dehydrogenase kinase activity by protein thiol-disulfide exchange.

    OpenAIRE

    Pettit, F H; Humphreys, J; Reed, L J

    1982-01-01

    Endogenous kinase activity of highly purified pyruvate dehydrogenase complex from bovine kidney is markedly inhibited by N-ethylmaleimide and by certain disulfides. Inhibition by disulfides is highly specific and is reversed by thiols. 5,5'-Dithiobis(2-nitrobenzoate) is the most potent inhibitor, showing significant inhibition at a concentration as low as 1 microM. Cystamine, oxidized glutathione, pantethine, lipoic acid, lipoamide, ergothionine, insulin, oxytocin, and vasopressin were ineffe...

  16. The integrity of the disulfide bond in a cyclic somatostatin analog during 99mtc complexation reactions

    International Nuclear Information System (INIS)

    Recent development of a variety of thiol-free chelating agents has facilitated the design of 99mTc-labeled somatostatin analogs suitable for receptor imaging of somatostatin-positive tumors. However, it remains ambiguous whether the disulfide bonds in cyclic peptides are stable during 99mTc complexation reactions, and contradictory results have been reported regarding the integrity of disulfide bonds in cyclic somatostatin analogs. To estimate the stability of the disulfide bond in a synthetic somatostatin analog at low peptide concentrations, [125I]I-RC-160, in which radioiodine was incorporated into the 3-Tyr residue, was synthesized and the integrity of the disulfide bond of the peptide was investigated in the presence of reducing agents such as ascorbic acid, dithionite, and stannous ions. The disulfide bond in [125I]I-RC-160 remained stable in the presence of ascorbic acid in boiling water. The disulfide bond was also stable when treated with stannous ions at concentrations sufficient to reduce 99mTc for complexation with a thiol-free chelating agent, bis(hydroxamamide) analog when the 99mTc complexation reaction was performed at room temperature. However, the disulfide bond of [125I]I-RC-160 was slightly cleaved in the presence of a small amount of stannous ions when the reaction was performed in boiling water. Treatment of [125I]I-RC-160 with dithionite in boiling water markedly reduced the disulfide bond of the parental peptide. These findings indicated that synthetic somatostatin analogs may be labeled with 99mTc with stannous ions as the reducing agent without impairing their structure after conjugation of thiol-free chelating agents that provide 99mTc chelates under mild reaction conditions

  17. An intact interchain disulfide bond is required for the neurotoxicity of tetanus toxin.

    Science.gov (United States)

    Schiavo, G; Papini, E; Genna, G; Montecucco, C

    1990-01-01

    Tetanus toxin is composed of a heavy chain (100 kDa) and a light chain (50 kDa) held together by a single interchain disulfide bridge. An additional intrachain disulfide is present in the carboxy-terminal part of the heavy chain. Reduction of the two disulfide bonds in tetanus toxin with both chemical and proteinaceous reducing agents was studied. Dithiothreitol and 2-mercaptoethanol cleaved both the inter- and intrachain disulfide bridges of the toxin, while glutathione and cysteine were ineffective. Specific reduction of the single interchain disulfide link was achieved with the thioredoxin-thioredoxin reductase system, thus indicating that this bond is exposed at the protein surface. Also, dead or permeabilized cells were able to reduce the toxin. Such reduced toxin bound to neuronal membranes as well as the native toxin but was not neurotoxic. These findings open the possibility that reduction by cytoplasmic agents released by dead cells contributes to detoxification of tetanus toxin. Moreover, together with the notion that the light chain is the active form of the toxin in the cytoplasm, these results suggest that the interchain disulfide bond of tetanus toxin plays a role in nerve cell penetration. Images PMID:2254033

  18. The effect of engineered disulfide bonds on the stability of Drosophila melanogaster acetylcholinesterase

    Directory of Open Access Journals (Sweden)

    Lamouroux Lucille

    2006-04-01

    Full Text Available Background Acetylcholinesterase is irreversibly inhibited by organophosphate and carbamate insecticides allowing its use in biosensors for detection of these insecticides. Drosophila acetylcholinesterase is the most sensitive enzyme known and has been improved by in vitro mutagenesis. However, its stability has to be improved for extensive utilization. Results To create a disulfide bond that could increase the stability of the Drosophila melanogaster acetylcholinesterase, we selected seven positions taking into account first the distance between Cβ of two residues, in which newly introduced cysteines will form the new disulfide bond and second the conservation of the residues in the cholinesterase family. Most disulfide bonds tested did not increase and even decreased the stability of the protein. However, one engineered disulfide bridge, I327C/D375C showed significant stability increase toward denaturation by temperature (170 fold at 50°C, urea, organic solvent and provided resistance to protease degradation. The new disulfide bridge links the N-terminal domain (first 356 aa to the C-terminal domain. The quantities produced by this mutant were the same as in wild-type flies. Conclusion Addition of a disulfide bridge may either stabilize or unstabilize proteins. One bond out of the 7 tested provided significant stabilisation.

  19. Stabilization of cyclohexanone monooxygenase by a computationally designed disulfide bond spanning only one residue.

    Science.gov (United States)

    van Beek, Hugo L; Wijma, Hein J; Fromont, Lucie; Janssen, Dick B; Fraaije, Marco W

    2014-01-01

    Enzyme stability is an important parameter in biocatalytic applications, and there is a strong need for efficient methods to generate robust enzymes. We investigated whether stabilizing disulfide bonds can be computationally designed based on a model structure. In our approach, unlike in previous disulfide engineering studies, short bonds spanning only a few residues were included. We used cyclohexanone monooxygenase (CHMO), a Baeyer-Villiger monooxygenase (BVMO) from Acinetobacter sp. NCIMB9871 as the target enzyme. This enzyme has been the prototype BVMO for many biocatalytic studies even though it is notoriously labile. After creating a small library of mutant enzymes with introduced cysteine pairs and subsequent screening for improved thermostability, three stabilizing disulfide bonds were identified. The introduced disulfide bonds are all within 12 Å of each other, suggesting this particular region is critical for unfolding. This study shows that stabilizing disulfide bonds do not have to span many residues, as the most stabilizing disulfide bond, L323C-A325C, spans only one residue while it stabilizes the enzyme, as shown by a 6 °C increase in its apparent melting temperature.

  20. Stabilization of cyclohexanone monooxygenase by a computationally designed disulfide bond spanning only one residue

    Directory of Open Access Journals (Sweden)

    Hugo L. van Beek

    2014-01-01

    Full Text Available Enzyme stability is an important parameter in biocatalytic applications, and there is a strong need for efficient methods to generate robust enzymes. We investigated whether stabilizing disulfide bonds can be computationally designed based on a model structure. In our approach, unlike in previous disulfide engineering studies, short bonds spanning only a few residues were included. We used cyclohexanone monooxygenase (CHMO, a Baeyer–Villiger monooxygenase (BVMO from Acinetobacter sp. NCIMB9871 as the target enzyme. This enzyme has been the prototype BVMO for many biocatalytic studies even though it is notoriously labile. After creating a small library of mutant enzymes with introduced cysteine pairs and subsequent screening for improved thermostability, three stabilizing disulfide bonds were identified. The introduced disulfide bonds are all within 12 Å of each other, suggesting this particular region is critical for unfolding. This study shows that stabilizing disulfide bonds do not have to span many residues, as the most stabilizing disulfide bond, L323C–A325C, spans only one residue while it stabilizes the enzyme, as shown by a 6 °C increase in its apparent melting temperature.

  1. One-Dimensional Electrical Contact to Molybdenum Disulfide

    Science.gov (United States)

    Yang, Zheng; Ra, Changho; Ahmed, Faisal; Lee, Daeyeong; Choi, Minsup; Liu, Xiaochi; Qu, Deshun; Yoo, Won Jong; Nano Device Processing Lab Team

    Molybdenum disulfide (MoS2) is one of the promising two-dimensional materials for future application in nano electronics, which has high carrier mobility, very good stability under atmosphere, proper band gap, etc. However, its application to electronic switching devices is hindered by Fermi level pinning at metal-MoS2 interfaces. Here, we experimentally demonstrate one-dimensional electrical contact to MoS2 formed via controllable plasma etching. We fabricated Al/MoS2 FET (n-type), Mo/MoS2 FET (n-type), and Pd/MoS2 FET (ambipolar). For Mo/MoS2 FET (n-type), on/off current ratio is around 108 and mobility is around 104 cm2/(Vs). By contrast, for Pd/MoS2 FET (ambipolar), on/off current ratio is around 108, hole mobility is ranged from 350 to 650 cm2/(Vs), and the mean free path of holes at 9K is around 23 nm. All the measured mobilities are evaluated by using two-terminal field-effect configuration. We can also achieve complementary logic gates with intrinsic MoS2/metal one-dimensional electrical contact.

  2. ALS-linked protein disulfide isomerase variants cause motor dysfunction.

    Science.gov (United States)

    Woehlbier, Ute; Colombo, Alicia; Saaranen, Mirva J; Pérez, Viviana; Ojeda, Jorge; Bustos, Fernando J; Andreu, Catherine I; Torres, Mauricio; Valenzuela, Vicente; Medinas, Danilo B; Rozas, Pablo; Vidal, Rene L; Lopez-Gonzalez, Rodrigo; Salameh, Johnny; Fernandez-Collemann, Sara; Muñoz, Natalia; Matus, Soledad; Armisen, Ricardo; Sagredo, Alfredo; Palma, Karina; Irrazabal, Thergiory; Almeida, Sandra; Gonzalez-Perez, Paloma; Campero, Mario; Gao, Fen-Biao; Henny, Pablo; van Zundert, Brigitte; Ruddock, Lloyd W; Concha, Miguel L; Henriquez, Juan P; Brown, Robert H; Hetz, Claudio

    2016-04-15

    Disturbance of endoplasmic reticulum (ER) proteostasis is a common feature of amyotrophic lateral sclerosis (ALS). Protein disulfide isomerases (PDIs) areERfoldases identified as possibleALSbiomarkers, as well as neuroprotective factors. However, no functional studies have addressed their impact on the disease process. Here, we functionally characterized fourALS-linked mutations recently identified in two majorPDIgenes,PDIA1 andPDIA3/ERp57. Phenotypic screening in zebrafish revealed that the expression of thesePDIvariants induce motor defects associated with a disruption of motoneuron connectivity. Similarly, the expression of mutantPDIs impaired dendritic outgrowth in motoneuron cell culture models. Cellular and biochemical studies identified distinct molecular defects underlying the pathogenicity of thesePDImutants. Finally, targetingERp57 in the nervous system led to severe motor dysfunction in mice associated with a loss of neuromuscular synapses. This study identifiesERproteostasis imbalance as a risk factor forALS, driving initial stages of the disease. PMID:26869642

  3. Aqueous Medium Synthesis Route for Randomly Stacked Molybdenum Disulfide

    Directory of Open Access Journals (Sweden)

    Pravas Kumar Panigrahi

    2013-01-01

    Full Text Available Synthesis of poorly crystalline, randomly oriented rag-like structures of molybdenum disulfide has been reported starting from aqueous solutions of ammonium molybdate, and thioacetamide in presence of sodium dodecyl sulfate via calcination of the amorphous precipitates, obtained through acidification of the in situ generated intermediate of ammonium tetrathiomolybdate. X-ray photoelectron spectroscopy, UV-visible spectroscopy, and X-ray diffraction of the calcined samples reveal the formation of single-phase MoS2, while the amorphous precipitates have been found to be a mixture of Mo2S5, MoS3, and a trace amount of H2MoS4. Highly folded and disordered layers of rag-like MoS2 have been confirmed through high-resolution transmission electron microscopy. The electrical conductivity for the cold pressed pellet of the MoS2 sample is found to be significantly higher than that of 2H-MoS2 and increases further on annealing.

  4. Template synthesis and characterization of molybdenum disulfide nanotubules

    International Nuclear Information System (INIS)

    Graphical abstract: The image is a SEM image of branched MoS2 nanotubes, which are prepared in AAO templates. It is obvious to observe the branch of MoS2 nanotubes (labeled by arrows), and it reflects the microcosmic morphologies of pores in templates. Highlights: → Large quantities of hollow MoS2 tubules. → Explanation for the formation of branched shape. → Explanation for the morphology of bamboo-like structure. -- Abstract: Molybdenum disulfide nanotubules were prepared by thermal decomposition of ammonium thiomolybdate ((NH4)2MoS4) precursors on anodized aluminum oxide template. Large quantities of hollow MoS2 nanotubules with the bamboo-like structure were obtained. The morphology and structures of MoS2 tubules were characterized by scanning electron microscopy, high-resolution transmission electron microscopy, energy dispersive spectroscopy, electron diffraction and optical absorption spectroscopy. MoS2 nanotubules completely reflected the three-dimensional structure of nanopores in template. The properties of Mo-S chemical bonds in lattice structure and the wetting state between porous surface and precursor have a great effect on the formation of sections in nanotubules, the ridges in the nanopores also play a very special role of this formation.

  5. Formation and stability of point defects in monolayer rhenium disulfide

    Science.gov (United States)

    Horzum, S.; ćakır, D.; Suh, J.; Tongay, S.; Huang, Y.-S.; Ho, C.-H.; Wu, J.; Sahin, H.; Peeters, F. M.

    2014-04-01

    Recently, rhenium disulfide (ReS2) monolayers were experimentally extracted by conventional mechanical exfoliation technique from as-grown ReS2 crystals. Unlike the well-known members of transition metal dichalcogenides (TMDs), ReS2 crystallizes in a stable distorted-1T structure and lacks an indirect to direct gap crossover. Here we present an experimental and theoretical study of the formation, energetics, and stability of the most prominent lattice defects in monolayer ReS2. Experimentally, irradiation with 3-MeV He+2 ions was used to break the strong covalent bonds in ReS2 flakes. Photoluminescence measurements showed that the luminescence from monolayers is mostly unchanged after highly energetic α particle irradiation. In order to understand the energetics of possible vacancies in ReS2 we performed systematic first-principles calculations. Our calculations revealed that the formation of a single sulfur vacancy has the lowest formation energy in both Re and S rich conditions and a random distribution of such defects are energetically more preferable. Sulfur point defects do not result in any spin polarization whereas the creation of Re-containing point defects induce magnetization with a net magnetic moment of 1-3μB. Experimentally observed easy formation of sulfur vacancies is in good agreement with first-principles calculations.

  6. Tension-Enhanced Hydrogen Evolution Reaction on Vanadium Disulfide Monolayer.

    Science.gov (United States)

    Pan, Hui

    2016-12-01

    Water electrolysis is an efficient way for hydrogen production. Finding efficient, cheap, and eco-friendly electrocatalysts is essential to the development of this technology. In the work, we present a first-principles study on the effects of tension on the hydrogen evolution reaction of a novel electrocatalyst, vanadium disulfide (VS2) monolayer. Two electrocatalytic processes, individual and collective processes, are investigated. We show that the catalytic ability of VS2 monolayer at higher hydrogen coverage can be efficiently improved by escalating tension. We find that the individual process is easier to occur in a wide range of hydrogen coverage and the collective process is possible at a certain hydrogen coverage under the same tension. The best hydrogen evolution reaction with near-zero Gibbs free energy can be achieved by tuning tension. We further show that the change of catalytic activity with tension and hydrogen coverage is induced by the change of free carrier density around the Fermi level, that is, higher carrier density, better catalytic performance. It is expected that tension can be a simple way to improve the catalytic activity, leading to the design of novel electrocatalysts for efficient hydrogen production from water electrolysis. PMID:26924817

  7. Novel high pressure structures and superconductivity of niobium disulfide

    International Nuclear Information System (INIS)

    Highlights: • At 26 GPa, NbS2 transits from the 2H structure to the novel I4/mmm structure. • The Nb and S atoms forms a new [NbS8] hexahedron unit in the I4/mmm-NbS2. • The I4/mmm-NbS2 exhibits a higher Tc than 2H-NbS2. • The higher Tc is resulted from the stronger electron–phonon coupling coefficients. - Abstract: We have investigated the pressure-induced phase transition and superconducting properties of niobium disulfide (NbS2) based on the density functional theory. The structures of NbS2 at pressures from 0 to 200 GPa were predicted using the multi-algorithm collaborative (MAC) structure prediction technique. The previously known 1T-, 2H-, and 3R-NbS2 were successfully reproduced. In addition, many metastable structures which are potential to be synthesized were also discovered. Based on the enthalpy calculations, we found that at 26 GPa NbS2 transits from the double-hexagonal (2H) structure to the tetragonal I4/mmm structure with a 10.6% volume reduction. The calculated elastic constants and phonon dispersion curves of I4/mmm-NbS2 confirm its mechanical and dynamical stability at high pressure. More interestingly, the coordination number of Nb in I4/mmm structure is eight which is larger than that in the traditional metal dichalcogenides, indicating a new type of bondings of Nb and S atoms. In the new Nb–S bondings, one Nb atom and neighboring eight S atoms form a [NbS8] hexahedron unit. Furthermore, I4/mmm-NbS2 exhibits a higher superconducting critical temperature than 2H-NbS2, as is resulted from the stronger electron–phonon coupling coefficients

  8. Nanoparticles synthesis of tungsten disulfide via AOT-based microemulsions

    International Nuclear Information System (INIS)

    Graphical abstract: A controlled synthesis of WS2 nanoparticles (most probably inorganic fullerene (IF)) via microemulsion was applied for the first time to prepare WS2 (7–12 nm) by acidification of the water cores of the AOT reverse microemulsion. Highlights: ► An innovative reverse microemulsion technique was developed for WS2 synthesis. ► WS2 nanoparticles were obtained with narrow size distribution in range of 7–12 nm. ► Operating cost of microemulsion was lower in contrast to quartz reactor method. ► WS2 morphology could be controlled to obtain highly active and selective catalysts. ► Lower size of WS2 in this study overcomes the shortcoming of quartz reactor method. -- Abstract: The tungsten disulfide (WS2) nanoparticles (most probably inorganic fullerene (IF)) with a narrow size distribution were synthesized by a reverse micelle technique for the first time. The particle size was controlled by varying water-to-surfactant molar ratio (W0), aging time and reagent concentration. The synthesized WS2 nanoparticles were characterized by zetasizer, UV–visible spectrophotometers and transmission electron microscopy (TEM). The WS2 nanoparticles with particle diameter size of 7–12 nm were obtained via 24 h aging time. The particle size was controlled by changing the aging time and molar ratio of water/surfactant. Doubling W0 increased the amount and particle size of WS2 by 22 and 26%, respectively. The effect of aging time in the range of 6–24 h was investigated and the complete disappearance of yellowish color at 24 h resulted in an optically clear solution, which was the indication of WS2 formation with 100% conversion of reactant ((NH4)2WS4) in the batch reactor.

  9. Preventing disulfide bond formation weakens non-covalent forces among lysozyme aggregates.

    Directory of Open Access Journals (Sweden)

    Vijay Kumar Ravi

    Full Text Available Nonnative disulfide bonds have been observed among protein aggregates in several diseases like amyotrophic lateral sclerosis, cataract and so on. The molecular mechanism by which formation of such bonds promotes protein aggregation is poorly understood. Here in this work we employ previously well characterized aggregation of hen eggwhite lysozyme (HEWL at alkaline pH to dissect the molecular role of nonnative disulfide bonds on growth of HEWL aggregates. We employed time-resolved fluorescence anisotropy, atomic force microscopy and single-molecule force spectroscopy to quantify the size, morphology and non-covalent interaction forces among the aggregates, respectively. These measurements were performed under conditions when disulfide bond formation was allowed (control and alternatively when it was prevented by alkylation of free thiols using iodoacetamide. Blocking disulfide bond formation affected growth but not growth kinetics of aggregates which were ∼50% reduced in volume, flatter in vertical dimension and non-fibrillar in comparison to control. Interestingly, single-molecule force spectroscopy data revealed that preventing disulfide bond formation weakened the non-covalent interaction forces among monomers in the aggregate by at least ten fold, thereby stalling their growth and yielding smaller aggregates in comparison to control. We conclude that while constrained protein chain dynamics in correctly disulfide bonded amyloidogenic proteins may protect them from venturing into partial folded conformations that can trigger entry into aggregation pathways, aberrant disulfide bonds in non-amyloidogenic proteins (like HEWL on the other hand, may strengthen non-covalent intermolecular forces among monomers and promote their aggregation.

  10. Synthesis and mechanistic studies of a mitomycin dimer containing an eight-membered cyclic disulfide.

    Science.gov (United States)

    Park, Hyun Jung; Kim, Jae Jin; Kim, Hyoung Rae; Lee, Eun Kyung; Kim, Eun Sook; Jeong, Choon Sik; Moon, Aree; Lee, Sang Hyup

    2011-07-01

    Dimeric DNA alkylating agents have drawn significant interest because these compounds are expected to provide at least two reactive sites and as a result, generate enhanced levels of DNA interstrand cross-link (DNA ISC) adducts compared to their monomeric agents. We report the synthesis and mechanistic studies of a novel mitomycin dimer, 7-N,7'-N'-(1″,2″-dithiocanyl-3″,8″-dimethylenyl)bismitomycin C (8) connected by an eight-membered cyclic disulfide. Mitomycins require prior activation (i.e., transformation to a good electrophile) for DNA adduction and therefore, 8 was aimed to undergo facile nucleophilic activation and produce enhanced levels of DNA ISC. At the core of this function lies a cyclic disulfide in 8. It was expected that disulfide cleavage by an appropriate nucleophile would successively produce two thiols that may trigger activation of two mitomycin rings in a dimer through intramolecular cyclization to quinine rings. Compound 8 was synthesized from mitomycin A (1) and the key intermediate, cyclic disulfide (11), along with the reference diol mitomycin 7-N,7'-N'-(2″,7″-dihydroxy-1″,8″-octanediyl)bismitomycin C (23) which does not contain the disulfide unit. We found that 8 underwent significantly enhanced nucleophilic activation in the presence of Et(3)P compared with 23, and that the disulfide unit in 8 played a key role for the nucleophilic activation. Based on these findings, we proposed a mechanism for nucleophilic activation of 8. We further demonstrated that 8 generated much higher levels of DNA ISC (94%) compared with 23 (4%) and 2 (3%) in the presence of Et(3)P (and L-DTT) leading to the conclusion that 8 is more efficient for DNA ISC processes than 23 and 2 due to the role of disulfide unit.

  11. Cytoplasmic glutathione redox status determines survival upon exposure to the thiol-oxidant 4,4'-dipyridyl disulfide

    DEFF Research Database (Denmark)

    López-Mirabal, H Reynaldo; Thorsen, Michael; Kielland-Brandt, Morten C;

    2007-01-01

    Dipyridyl disulfide (DPS) is a highly reactive thiol oxidant that functions as electron acceptor in thiol-disulfide exchange reactions. DPS is very toxic to yeasts, impairing growth at low micromolar concentrations. The genes TRX2 (thioredoxin), SOD1 (superoxide dismutase), GSH1 (gamma-glutamyl-c...... sensitivity. DPS seems to induce a specific disulfide stress, where an increase in the cytoplasmic/nuclear GSSG/GSH ratio results in putative DPS target(s) becoming sensitive to DPS. Udgivelsesdato: 2007-May...

  12. Green polymer chemistry: Synthesis of poly(disulfide) polymers and networks

    Science.gov (United States)

    Rosenthal-Kim, Emily Quinn

    The disulfide group is unique in that it presents a covalent bond that is easily formed and cleaved under certain biological conditions. While the ease of disulfide bond cleavage is often harnessed as a method of biodegradation, the ease of disulfide bond formation as a synthetic strategy is often overlooked. The objective this research was to synthesize poly(disulfide) polymers and disulfide crosslinked networks from a green chemistry approach. The intent of the green chemistry approach was to take advantage of the mild conditions applicable to disulfide bond synthesis from thiols. With anticipated use as biomaterials, it was also desired that the polymer materials could be degraded under biological conditions. Here, a new method of poly(disulfide) polymer synthesis is introduced which was inspired by the reaction conditions and reagents found in Nature. Ambient temperatures and aqueous mixtures were used in the new method. Hydrogen peroxide, one of the Nature's most powerful oxidizing species was used as the oxidant in the new polymerization reaction. The dithiol monomer, 3,6-dioxa-1,8-octanedithiol was first solubilized in triethylamine, which activated the thiol groups and made the monomer water soluble. At room temperature, the organic dithiol/amine solution was then mixed with dilute aqueous hydrogen peroxide (3% by weight) to make the poly(disulfide) polymers. The presence of a two phase system (organic and aqueous phases) was critical to the polymerization reaction. As the reaction progresses, a third, polymer phase appeared. At ambient temperatures and above, this phase separated from the reaction mixture and the polymer product was easily removed from the reaction solution. These polymers reach Mn > 250,000 g/mol in under two hours. Molecular weight distributions were between 1.5 and 2.0. Reactions performed in an ice bath which remain below room temperature contain high molecular weight polymers with Mn ≈ 120,000 g/mol and have a molecular weight

  13. Molybdenum disulfide (MoS2) nanoflakes as inherently electroactive labels for DNA hybridization detection.

    Science.gov (United States)

    Loo, Adeline Huiling; Bonanni, Alessandra; Ambrosi, Adriano; Pumera, Martin

    2014-10-21

    The detection of specific DNA sequences plays a critical role in the areas of medical diagnostics, environmental monitoring, drug discovery and food safety. This has therefore become a strong driving force behind the ever-increasing demand for simple, cost-effective, highly sensitive and selective DNA biosensors. In this study, we report for the first time, a novel approach for the utilization of molybdenum disulfide nanoflakes, a member of the transition metal dichalcogenides family, in the detection of DNA hybridization. Herein, molybdenum disulfide nanoflakes serve as inherently electroactive labels, with the inherent oxidation peak exploited as the analytical signal. The principle of detection is based on the differential affinity of molybdenum disulfide nanoflakes towards single-stranded DNA and double-stranded DNA. The employment of transition metal dichalcogenide nanomaterials for sensing and biosensing purposes represents an upcoming research area which holds great promise. Hence, our findings are anticipated to have significant contributions towards the fabrication of future DNA biosensors. PMID:25177907

  14. Electrochemical reduction of disulfide-containing proteins for hydrogen/deuterium exchange monitored by mass spectrometry

    DEFF Research Database (Denmark)

    Mysling, Simon; Salbo, Rune; Ploug, Michael;

    2014-01-01

    some challenges in using electrochemical reduction in HDX-MS analyses and provide possible conditions to attenuate these limitations. For example, high salt concentrations hamper disulfide bond reduction, necessitating additional dilution of the sample with aqueous acidic solution at quench conditions.......Characterization of disulfide bond-containing proteins by hydrogen/deuterium exchange monitored by mass spectrometry (HDX-MS) requires reduction of the disulfide bonds under acidic and cold conditions, where the amide hydrogen exchange reaction is quenched (pH 2.5, 0 °C). The reduction typically...... requires a high concentration (>200 mM) of the chemical reducing agent Tris(2-carboxyethyl)phosphine (TCEP) as its reduction rate constant is decreased at low pH and temperature. Serious adverse effects on chromatographic and mass spectrometric performances have been reported when using high concentrations...

  15. Glutathione, glutathione disulfide, and S-glutathionylated proteins in cell cultures.

    Science.gov (United States)

    Giustarini, Daniela; Galvagni, Federico; Tesei, Anna; Farolfi, Alberto; Zanoni, Michele; Pignatta, Sara; Milzani, Aldo; Marone, Ilaria M; Dalle-Donne, Isabella; Nassini, Romina; Rossi, Ranieri

    2015-12-01

    The analysis of the global thiol-disulfide redox status in tissues and cells is a challenging task since thiols and disulfides can undergo artificial oxido-reductions during sample manipulation. Because of this, the measured values, in particular for disulfides, can have a significant bias. Whereas this methodological problem has already been addressed in samples of red blood cells and solid tissues, a reliable method to measure thiols and disulfides in cell cultures has not been previously reported. Here, we demonstrate that the major artifact occurring during thiol and disulfide analysis in cultured cells is represented by glutathione disulfide (GSSG) and S-glutathionylated proteins (PSSG) overestimation, due to artificial oxidation of glutathione (GSH) during sample manipulation, and that this methodological problem can be solved by the addition of N-ethylmaleimide (NEM) immediately after culture medium removal. Basal levels of GSSG and PSSG in different lines of cultured cells were 3-5 and 10-20 folds higher, respectively, when the cells were processed without NEM. NEM pre-treatment also prevented the artificial reduction of disulfides that occurs during the pre-analytical phase when cells are exposed to an oxidant stimulus. In fact, in the absence of NEM, after medium removal, GSH, GSSG and PSSG levels restored their initial values within 15-30 min, due to the activity of reductases and the lack of the oxidant. The newly developed protocol was used to measure the thiol-disulfide redox status in 16 different line cells routinely used for biomedical research both under basal conditions and after treatment with disulfiram, a thiol-specific oxidant (0-200 μM concentration range). Our data indicate that, in most cell lines, treatment with disulfiram affected the levels of GSH and GSSG only at the highest concentration. On the other hand, PSSG levels increased significantly also at the lower concentrations of the drug, and the rise was remarkable (from 100 to 1000

  16. Der Thiol:Disulfid-Redox Metabolismus und der Blaulichtrezeptor Lmo0799 von Listeria monocytogenes

    OpenAIRE

    Ondrusch, Nicolai

    2010-01-01

    Der Thiol-Redox-Metabolismus, der in allen lebenden Zellen zu finden ist, wirkt oxidativem Stress entgegen. Des Weiteren dient er auch der Aufrechterhaltung der intrazellulären Thiol:Disulfid-Balance, die wiederum für die Funktion vieler Proteine essentiell ist. Auch stellt er Reduktionsäquivalente für die Produktion von Desoxyribonucleotiden für die DNA-Synthese bereit und hilft oxidierte Proteine zu reparieren. Der Thiol:Disulfid-Redox-Metabolismus (TDRM) unterscheidet sich von anderen meta...

  17. Probing the structure of human protein disulfide isomerase by chemical cross-linking combined with mass spectrometry

    DEFF Research Database (Denmark)

    Peng, Li; Rasmussen, Morten Ib; Chailyan, Anna;

    2014-01-01

    Protein disulfide-isomerase (PDI) is a four-domain flexible protein that catalyzes the formation of disulfide bonds in the endoplasmic reticulum. Here we have analyzed native PDI purified from human placenta by chemical cross-linking followed by mass spectrometry (CXMS). In addition to PDI the sa...

  18. Role of disulfide linkage in action of bis(dialkylaminethiocarbonyl)disulfides as potent double-Edged microbicidal spermicide: Design, synthesis and biology.

    Science.gov (United States)

    Lal, Nand; Jangir, Santosh; Bala, Veenu; Mandalapu, Dhanaraju; Sarswat, Amit; Kumar, Lalit; Jain, Ashish; Kumar, Lokesh; Kushwaha, Bhavana; Pandey, Atindra K; Krishna, Shagun; Rawat, Tara; Shukla, Praveen K; Maikhuri, Jagdamba P; Siddiqi, Mohammad I; Gupta, Gopal; Sharma, Vishnu L

    2016-06-10

    Trichomoniasis and candidiasis are amongst the most common morbidity-causing reproductive tract infections, generally treated by Metronidazole and Fluconazole respectively. Poor vaginal efficacy, drug-resistance and non-spermicidal nature limit their use as topical microbicidal contraceptives. Bis(dialkylaminethiocarbonyl)disulfides (4-38) were designed as dually active, non-surfactant molecules capable of eliminating Trichomonas vaginalis and Candida strains as well as irreversibly immobilizing 100% human sperm instantly, at doses non-cytotoxic to human cervical epithelial cells and vaginal microflora in vitro. Compounds 12, 16, 17 were fifty times more active than nonoxynol-9, OTC vaginal spermicide, and compounds 12 and 17 have shown remarkable in vivo activity in rabbit model. Most promising compound 17 has shown promise for further development as a double-edged vaginal microbicide due to their improved activity and safety along with notable in vivo trichomonicidal activity. Role of disulfide group was established by loss of spermicidal activity on chemical modifications (39-56). These disulfides might be targeting thiol groups present over cell membrane of human sperm and Trichomonas as shown by fluorescence labeling of free thiols. PMID:27084496

  19. Two-Dimensional Structure of Disulfides and Thiols on Gold(111)

    NARCIS (Netherlands)

    Nelles, Gabriele; Schönherr, Holger; Jaschke, Manfred; Wolf, Heiko; Schaub, Matthias; Kuther, Jörg; Tremel, Wolfgang; Bamberg, Ernst; Ringsdorf, Helmut; Butt, Hans-Jürgen

    1998-01-01

    In order to find factors which determine the two-dimensional structure of self-assembled monolayers (SAMs), several classes of thiols and disulfides on gold (111) have been investigated by atomic force microscopy (AFM). SAMs were formed from a series of symmetrical and asymmetrical diethylalkanoate

  20. Protein disulfide isomerase homolog TrPDI2 contributing to cellobiohydrolase production in Trichoderma reesei.

    Science.gov (United States)

    Wang, Guokun; Lv, Pin; He, Ronglin; Wang, Haijun; Wang, Lixian; Zhang, Dongyuan; Chen, Shulin

    2015-09-01

    The majority of the cysteine residues in the secreted proteins form disulfide bonds via protein disulfide isomerase (PDI)-mediated catalysis, stabilizing the enzyme activity. The role of PDI in cellulase production is speculative, as well as the possibility of PDI as a target for improving enzyme production efficiency of Trichoderma reesei, a widely used producer of enzyme for the production of lignocellulose-based biofuels and biochemicals. Here, we report that a PDI homolog, TrPDI2 in T. reesei exhibited a 36.94% and an 11.81% similarity to Aspergillus niger TIGA and T. reesei PDI1, respectively. The capability of TrPDI2 to recover the activity of reduced and denatured RNase by promoting refolding verified its protein disulfide isomerase activity. The overexpression of Trpdi2 increased the secretion and the activity of CBH1 at the early stage of cellulase induction. In addition, both the expression level and redox state of TrPDI2 responded to cellulase induction in T. reesei, providing sustainable oxidative power to ensure cellobiohydrolase maturation and production. The results suggest that TrPDI2 may contribute to cellobiohydrolase secretion by enhancing the capability of disulfide bond formation, which is essential for protein folding and maturation. PMID:26138396

  1. Inactivation of barley limit dextrinase inhibitor by thioredoxin-catalysed disulfide reduction

    DEFF Research Database (Denmark)

    Jensen, Johanne Mørch; Hägglund, Per; Christensen, Hans Erik Mølager;

    2012-01-01

    Barley limit dextrinase (LD) that catalyses hydrolysis of α-1,6 glucosidic linkages in starch-derived dextrins is inhibited by limit dextrinase inhibitor (LDI) found in mature seeds. LDI belongs to the chloroform/methanol soluble protein family (CM-protein family) and has four disulfide bridges a...

  2. Characterization of a foldase, protein disulfide isomerase A, in the protein secretory pathway of Aspergillus niger

    NARCIS (Netherlands)

    Ngiam, C.; Jeenes, D.J.; Punt, P.J.; Hondel, C.A.M.J.J. van den; Archer, D.B.

    2000-01-01

    Protein disulfide isomerase (PDI) is important in assisting the folding and maturation of secretory proteins in eukaryotes. A gene, pdiA, encoding PDIA was previously isolated from Aspergillus niger, and we report its functional characterization here. Functional analysis of PDIA showed that it catal

  3. Dissecting the role of disulfide bonds on the amyloid formation of insulin

    International Nuclear Information System (INIS)

    Highlights: ► We dissect how individual disulfide bond affects the amyloidogenicity of insulin. ► A controlled reduction system for insulin is established in this study. ► Disulfide breakage is associated with unfolding and increased amyloidogenicity. ► Breakage of A6-A11 is associated with significantly increased cytotoxicity. ► Analogs without A6-A11 have a higher potency to form high order toxic oligomers. -- Abstract: Disulfide bonds play a critical role in the stability and folding of proteins. Here, we used insulin as a model system, to investigate the role of its individual disulfide bond during the amyloid formation of insulin. Tris(2-carboxyethyl)phosphine (TCEP) was applied to reduce two of the three disulfide bonds in porcine insulin and the reduced disulfide bonds were then alkylated by iodoacetamide. Three disulfide bond-modified insulin analogs, INS-2 (lack of A6-A11), INS-3 (lack of A7-B7) and INS-6 (lack of both A6-A11 and A7-B7), were obtained. Far-UV circular dichroism (CD) spectroscopy results indicated that the secondary structure of INS-2 was the closest to insulin under neutral conditions, followed by INS-3 and INS-6, whereas in an acidic solution all analogs were essentially unfolded. To test how these modifications affect the amyloidogenicity of insulin, thioflavin-T (ThT) fluorescence and transmission electronic microscopy (TEM) were performed. Our results showed that all analogs were more prone to aggregation than insulin, with the order of aggregation rates being INS-6 > INS-3 > INS-2. Cross-linking of unmodified proteins (PICUP) assay results showed that analogs without A6-A11 (INS-2 and INS-6) have a higher potential for oligomerization than insulin and INS-3, which is accompanied with a higher cytotoxicity as the hemolytic assays of human erythrocytes suggested. The results indicated that breakage of A7-B7 induced more unfolding of the insulin structure and a higher amyloidogenicity than breakage of A6-A11, but breakage of A6

  4. Dissecting the role of disulfide bonds on the amyloid formation of insulin

    Energy Technology Data Exchange (ETDEWEB)

    Li, Yang; Gong, Hao [Tongji School of Pharmacy, Huazhong University of Science and Technology, Wuhan 430030 (China); Sun, Yue [College of Life Sciences, Wuhan University, Wuhan 430072 (China); Yan, Juan; Cheng, Biao; Zhang, Xin [Tongji School of Pharmacy, Huazhong University of Science and Technology, Wuhan 430030 (China); Huang, Jing [College of Life Sciences, Wuhan University, Wuhan 430072 (China); Yu, Mengying; Guo, Yu [Tongji School of Pharmacy, Huazhong University of Science and Technology, Wuhan 430030 (China); Zheng, Ling, E-mail: lzheng217@hotmail.com [College of Life Sciences, Wuhan University, Wuhan 430072 (China); Huang, Kun, E-mail: kunhuang2008@hotmail.com [Tongji School of Pharmacy, Huazhong University of Science and Technology, Wuhan 430030 (China); Centre for Biomedicine Research, Wuhan Institutes of Biotechnology, Wuhan 430070 (China)

    2012-06-29

    Highlights: Black-Right-Pointing-Pointer We dissect how individual disulfide bond affects the amyloidogenicity of insulin. Black-Right-Pointing-Pointer A controlled reduction system for insulin is established in this study. Black-Right-Pointing-Pointer Disulfide breakage is associated with unfolding and increased amyloidogenicity. Black-Right-Pointing-Pointer Breakage of A6-A11 is associated with significantly increased cytotoxicity. Black-Right-Pointing-Pointer Analogs without A6-A11 have a higher potency to form high order toxic oligomers. -- Abstract: Disulfide bonds play a critical role in the stability and folding of proteins. Here, we used insulin as a model system, to investigate the role of its individual disulfide bond during the amyloid formation of insulin. Tris(2-carboxyethyl)phosphine (TCEP) was applied to reduce two of the three disulfide bonds in porcine insulin and the reduced disulfide bonds were then alkylated by iodoacetamide. Three disulfide bond-modified insulin analogs, INS-2 (lack of A6-A11), INS-3 (lack of A7-B7) and INS-6 (lack of both A6-A11 and A7-B7), were obtained. Far-UV circular dichroism (CD) spectroscopy results indicated that the secondary structure of INS-2 was the closest to insulin under neutral conditions, followed by INS-3 and INS-6, whereas in an acidic solution all analogs were essentially unfolded. To test how these modifications affect the amyloidogenicity of insulin, thioflavin-T (ThT) fluorescence and transmission electronic microscopy (TEM) were performed. Our results showed that all analogs were more prone to aggregation than insulin, with the order of aggregation rates being INS-6 > INS-3 > INS-2. Cross-linking of unmodified proteins (PICUP) assay results showed that analogs without A6-A11 (INS-2 and INS-6) have a higher potential for oligomerization than insulin and INS-3, which is accompanied with a higher cytotoxicity as the hemolytic assays of human erythrocytes suggested. The results indicated that breakage of A7

  5. Close collisions in the two-dimensional Raman response of liquid carbon disulfide

    NARCIS (Netherlands)

    Jansen, TLC; Duppen, K; Snijders, Jaap

    2003-01-01

    The fifth-order 2D Raman response of a liquid is calculated taking all possible interaction induced effects into account. Next to dipole-induced dipole interactions, close collision effects due to induced multipoles and electron overlap are found to give a significant contribution to the response of

  6. Human Sperm Chromosome Analysis—Study on Human Sperm Chromosome Mutagenesis Induced by Carbon Disulfide

    Institute of Scientific and Technical Information of China (English)

    LEJUN-YI; FUXIAO-MIN

    1996-01-01

    The aim of this study was to investigate the effect CS2 of on human sperm chromosomal aberration.The human sperm/hamster egg fusion techniquse was used to analyze 203 human sperm chromosome complement form 9 healthy volunteers.The incidence of numerical aberration was 1.0%,and that of structural chromosome aberration was 5.9% and total abnormalities was 6.9%.Structural aberrations consisted of breaks,deletions, centric rings,fragments,and chromatid exchange.The results from high concentration group(10μmol·L-1 CS2)showed that the incidence of chromosomal aberration rate was significantly higher than that of the control group.The results indicate that high concentration of CS2 might directly cause mutatenesis f the germ cell.

  7. Modeling of a trickling bioreactor for the simultaneous removal of hydrogen sulfide and carbon disulfide

    Energy Technology Data Exchange (ETDEWEB)

    Lobo, R.; Viveros-Garcia, T.; Revah, S. [Universidad Autonoma Metropolitana-Iztapalapa, Mexico D.F. (Mexico)

    1996-12-31

    A model is presented for a trickle biofilm reactor based on liquid falling film hydrodynamics and linear residence times. It is shown that mass transfer resistance is high. Performance of pseudo homogeneous and heterogeneous models is discussed. It is shown that the use of superficial residence times reduces the heterogeneous model to a pseudo homogeneous one and masks parametric response. 8 refs., 2 figs., 1 tab.

  8. An analysis of a trickle-bed bioreactor: Carbon disulfide removal

    Energy Technology Data Exchange (ETDEWEB)

    Lobo, R.; Revah, S.; Viveros-Garcia, T. [Univ. Autonoma Metropolitana-Iztapalapa (Mexico). Dept. de Ingenieria de Procesos e Hidraulica

    1999-04-05

    An analysis of the local processes occurring in a trickle-bed bioreactor (TBB) with a first-order bioreaction shows that the identification of the TBB operating regime requires knowledge of the substrate concentration in the liquid phase. If the substrate liquid concentration is close to 0, the rate-controlling step is mass transfer at the gas-liquid interface; when it is close to the value in equilibrium with the gas phase, the controlling step is the phenomena occurring in the biofilm, CS{sub 2} removal rate data obtained in a TBB with a Thiobacilii consortia biofilm are analyzed to obtain the mass transfer and kinetic parameters, and to show that the bioreactor operates in a regime mainly controlled by mass transfer. A TBB model with two experimentally determined parameters is developed and used to show how the bioreactor size depends on the rate-limiting step, the absorption factor, the substrate fractional conversion, and on the gas and liquid contact pattern. Under certain conditions, the TBB size is independent of the flowing phases` contact pattern. The model effectively describes substrate gas and liquid concentration data for mass transfer and biodegradation rate controlled processes.

  9. Study on the reaction of carbon disulfide with hydroxyl radical in aqueous solution

    Institute of Scientific and Technical Information of China (English)

    FANG Haojie; OUYANG Bin; QIN Yan; DONG Wenbo; HOU Huiqi

    2005-01-01

    The laser flash photolysis technique is employed to investigate the reaction mechanism of CS2 with ·OH in the nitrogen-saturated aqueous solution. By comparison of the transient absorption spectra obtained at different phases and pHs and through the addition of proper radical cation scavenger, CS2 is determined to react with ·OH to form ·CS2OH adduct, instead of the CS2+ radical cation. At pH 1-5, ·CS2OH decomposes into COS and ·HS, while at pH>5, it further reacts with OH- to form CS2O-. The temperature dependent kinetics for the reaction CS2 + ·OH →·CS2OH is also reported in this paper with an estimated activation energy of (26.9±1.0) kJ·mol1.

  10. Transport Measurements in the Mixed Ion-Electron Conductor Copper(x) Carbon-Disulfide

    Science.gov (United States)

    Kuo, Hung-Jen

    (sigma)(,i), (sigma)(,e), and the chemical diffusion coefficient, (')D, of highly-disordered Cu(,x)CS(,2) were investigated using a dc 4-lead technique employing Pt electrodes. The experiments were performed at various copper concentrations from x = 2.87 to 3.60 and various temperatures from 260 K to 350 K. The results were interpreted by Yokota's and ionic hopping diffusion theories. (sigma)(,i) and (sigma)(,e) are comparable at room temperature, 4.18 x 10('-3) (OMEGA)('-1)cm('-1) and 1.55 x 10('-3) (OMEGA)('-1)cm('-1) respectively at X = 3.60 and 300 K. Both (sigma)(,i) and (sigma)(,e) follow a simple Arrhenius form with activation energies (TURN)0.40 eV and (TURN)0.29 eV respectively. The exponential dependence of (sigma)(,i) on X is explained in terms of the activation entropy associated with the motion of ions. Electronic conduction is by hopping. Results show that it is reasonable to assume that all the copper ions are mobile. The mobility and the diffusivity of copper ions were found to be 0.71 x 10('-6) cm('2)V(' -1)sec('-1) and 1.83 x 10('-8) cm('2)/sec respectively at X = 3.6 and 300 K. The diffusivity is much less than the chemical diffusion coefficient evaluated from the diffusion time constant, (')D = 0.829 x 10('-5) cm('2)/sec at X = 3.60 and 300 K. This is because of a large enhancement factor W (TURN) 453, or a large (PAR-DIFF)m(,e)/(PAR-DIFF)N. The change in galvanic cell potential E with X, -(PAR-DIFF)E/(PAR -DIFF)X, calculated from the measurements of (sigma)(,i), (sigma)(,e), and (')D, is 14 Volt.

  11. Hyperstabilization of Tetrameric Bacillus sp. TB-90 Urate Oxidase by Introducing Disulfide Bonds through Structural Plasticity.

    Science.gov (United States)

    Hibi, Takao; Kume, Asami; Kawamura, Akie; Itoh, Takafumi; Fukada, Harumi; Nishiya, Yoshiaki

    2016-02-01

    Bacillus sp. TB-90 urate oxidase (BTUO) is one of the most thermostable homotetrameric enzymes. We previously reported [Hibi, T., et al. (2014) Biochemistry 53, 3879-3888] that specific binding of a sulfate anion induced thermostabilization of the enzyme, because the bound sulfate formed a salt bridge with two Arg298 residues, which stabilized the packing between two β-barrel dimers. To extensively characterize the sulfate-binding site, Arg298 was substituted with cysteine by site-directed mutagenesis. This substitution markedly increased the protein melting temperature by ∼ 20 °C compared with that of the wild-type enzyme, which was canceled by reduction with dithiothreitol. Calorimetric analysis of the thermal denaturation suggested that the hyperstabilization resulted from suppression of the dissociation of the tetramer into the two homodimers. The crystal structure of R298C at 2.05 Å resolution revealed distinct disulfide bond formation between the symmetrically related subunits via Cys298, although the Cβ distance between Arg298 residues of the wild-type enzyme (5.4 Å apart) was too large to predict stable formation of an engineered disulfide cross-link. Disulfide bonding was associated with local disordering of interface loop II (residues 277-300), which suggested that the structural plasticity of the loop allowed hyperstabilization by disulfide formation. Another conformational change in the C-terminal region led to intersubunit hydrogen bonding between Arg7 and Asp312, which probably promoted mutant thermostability. Knowledge of the disulfide linkage of flexible loops at the subunit interface will help in the development of new strategies for enhancing the thermostabilization of multimeric proteins. PMID:26739254

  12. Identification of a disulfide bridge important for transport function of SNAT4 neutral amino acid transporter.

    Directory of Open Access Journals (Sweden)

    Rugmani Padmanabhan Iyer

    Full Text Available SNAT4 is a member of system N/A amino acid transport family that primarily expresses in liver and muscles and mediates the transport of L-alanine. However, little is known about the structure and function of the SNAT family of transporters. In this study, we showed a dose-dependent inhibition in transporter activity of SNAT4 with the treatment of reducing agents, dithiothreitol (DTT and Tris(2-carboxyethylphosphine (TCEP, indicating the possible involvement of disulfide bridge(s. Mutation of residue Cys-232, and the two highly conserved residues Cys-249 and Cys-321, compromised the transport function of SNAT4. However, this reduction was not caused by the decrease of SNAT4 on the cell surface since the cysteine-null mutant generated by replacing all five cysteines with alanine was equally capable of being expressed on the cell surface as wild-type SNAT4. Interestingly, by retaining two cysteine residues, 249 and 321, a significant level of L-alanine uptake was restored, indicating the possible formation of disulfide bond between these two conserved residues. Biotinylation crosslinking of free thiol groups with MTSEA-biotin provided direct evidence for the existence of a disulfide bridge between Cys-249 and Cys-321. Moreover, in the presence of DTT or TCEP, transport activity of the mutant retaining Cys-249 and Cys-321 was reduced in a dose-dependent manner and this reduction is gradually recovered with increased concentration of H2O2. Disruption of the disulfide bridge also decreased the transport of L-arginine, but to a lesser degree than that of L-alanine. Together, these results suggest that cysteine residues 249 and 321 form a disulfide bridge, which plays an important role in substrate transport but has no effect on trafficking of SNAT4 to the cell surface.

  13. Carbon-Dot-Based Nanosensors for the Detection of Intracellular Redox State.

    Science.gov (United States)

    Liu, Ye; Tian, Ye; Tian, Yefei; Wang, Yajun; Yang, Wuli

    2015-11-25

    Carbon-dot-based nanosensors are prepared through sequentially assembling a polymer/carbon dot multilayer shell on mesoporous silica nanoparticles with different crosslinking densities of disulfide bonds; they can be utilized to evaluate the gluthathione (GSH) concentration. In vitro cell assays demonstrate the feasibility of using such nanosensors in evaluating the intracellular redox state of different cells. PMID:26450796

  14. Free-standing hierarchically sandwich-type tungsten disulfide nanotubes/graphene anode for lithium-ion batteries.

    Science.gov (United States)

    Chen, Renjie; Zhao, Teng; Wu, Weiping; Wu, Feng; Li, Li; Qian, Ji; Xu, Rui; Wu, Huiming; Albishri, Hassan M; Al-Bogami, A S; El-Hady, Deia Abd; Lu, Jun; Amine, Khalil

    2014-10-01

    Transition metal dichalcogenides (TMD), analogue of graphene, could form various dimensionalities. Similar to carbon, one-dimensional (1D) nanotube of TMD materials has wide application in hydrogen storage, Li-ion batteries, and supercapacitors due to their unique structure and properties. Here we demonstrate the feasibility of tungsten disulfide nanotubes (WS2-NTs)/graphene (GS) sandwich-type architecture as anode for lithium-ion batteries for the first time. The graphene-based hierarchical architecture plays vital roles in achieving fast electron/ion transfer, thus leading to good electrochemical performance. When evaluated as anode, WS2-NTs/GS hybrid could maintain a capacity of 318.6 mA/g over 500 cycles at a current density of 1A/g. Besides, the hybrid anode does not require any additional polymetric binder, conductive additives, or a separate metal current-collector. The relatively high density of this hybrid is beneficial for high capacity per unit volume. Those characteristics make it a potential anode material for light and high-performance lithium-ion batteries. PMID:25163033

  15. Universal low-temperature MWCNT-COOH-based counter electrode and a new thiolate/disulfide electrolyte system for dye-sensitized solar cells.

    Science.gov (United States)

    Hilmi, Abdulla; Shoker, Tharallah A; Ghaddar, Tarek H

    2014-06-11

    A new thiolate/disulfide organic-based electrolyte system composed of the tetrabutylammonium salt of 2-methyl-5-trifluoromethyl-2H-[1,2,4]triazole-3-thiol (S(-)) and its oxidized form 3,3'-dithiobis(2-methyl-5-trifluoromethyl-2H-[1,2,4]triazole) (DS) has been formulated and used in dye-sensitized solar cells (DSSCs). The electrocatalytic activity of different counter electrodes (CEs) has been evaluated by means of measuring J-V curves, cyclic voltammetry, Tafel plots, and electrochemical impedance spectroscopy. A stable and low-temperature CE based on acid-functionalized multiwalled carbon nanotubes (MWCNT-COOH) was investigated with our S(-)/DS, I(-)/I3(-), T(-)/T2, and Co(II/III)-based electrolyte systems. The proposed CE showed superb electrocatalytic activity toward the regeneration of the different electrolytes. In addition, good stability of solar cell devices based on the reported electrolyte and CE was shown.

  16. A second disulfide bridge from the N-terminal domain to extracellular loop 2 dampens receptor activity in GPR39

    DEFF Research Database (Denmark)

    Storjohann, Laura; Holst, Birgitte; Schwartz, Thue W

    2008-01-01

    . Disruption of the nonconserved disulfide bridge by mutagenesis led to an increase in the Zn (2+) potency. This phenotype, with an approximate 10-fold increase in agonist potency and a slight increase in E max, was mimicked by treatment of the wild-type receptor with TCEP at low concentrations, which had no......A highly conserved feature across all families of 7TM receptors is a disulfide bridge between a Cys residue located at the extracellular end of transmembrane segment III (TM-III) and one in extracellular loop 2 (ECL-2). The zinc sensor GPR39 contains four Cys residues in the extracellular domains....... By using mutagenesis, treatment with the reducing agent TCEP, and a labeling procedure for free sulfhydryl groups, we identify the pairing of these Cys residues in two disulfide bridges: the prototypical bridge between Cys (108) in TM-III and Cys (210) in ECL-2 and a second disulfide bridge...

  17. On-Line Electrochemical Reduction of Disulfide Bonds: Improved FTICR-CID and -ETD Coverage of Oxytocin and Hepcidin

    Science.gov (United States)

    Nicolardi, Simone; Giera, Martin; Kooijman, Pieter; Kraj, Agnieszka; Chervet, Jean-Pierre; Deelder, André M.; van der Burgt, Yuri E. M.

    2013-12-01

    Particularly in the field of middle- and top-down peptide and protein analysis, disulfide bridges can severely hinder fragmentation and thus impede sequence analysis (coverage). Here we present an on-line/electrochemistry/ESI-FTICR-MS approach, which was applied to the analysis of the primary structure of oxytocin, containing one disulfide bridge, and of hepcidin, containing four disulfide bridges. The presented workflow provided up to 80 % (on-line) conversion of disulfide bonds in both peptides. With minimal sample preparation, such reduction resulted in a higher number of peptide backbone cleavages upon CID or ETD fragmentation, and thus yielded improved sequence coverage. The cycle times, including electrode recovery, were rapid and, therefore, might very well be coupled with liquid chromatography for protein or peptide separation, which has great potential for high-throughput analysis.

  18. Tetracene dicarboxylic imide and its disulfide: synthesis of ambipolar organic semiconductors for organic photovoltaic cells.

    Science.gov (United States)

    Okamoto, Toshihiro; Suzuki, Tsuyoshi; Tanaka, Hideyuki; Hashizume, Daisuke; Matsuo, Yutaka

    2012-01-01

    We have designed and synthesized a new donor/acceptor-type tetracene derivative by the introduction of dicarboxylic imide and disulfide groups as electron-withdrawing and -donating units, respectively. The prepared compounds, tetracene dicarboxylic imide (TI) and its disulfide (TIDS) have high chemical and electrochemical stability as well as long-wavelength absorptions of up to 886 nm in the thin films. The crystal packing structure of TIDS molecules features face-to-face π-stacking, derived from dipole-dipole interactions. Notably, TIDS exhibited ambipolar properties of both electron-donating and -accepting natures in p-n and p-i-n heterojunction organic thin-film photovoltaic devices. Accordingly, TI and TIDS are expected to be promising compounds for designing new organic semiconductors.

  19. Conversion of a disulfide bond into a thioacetal group during echinomycin biosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    Hotta, Kinya; Keegan, Ronan M.; Ranganathan, Soumya; Fang, Minyi; Bibby, Jaclyn; Winn, Martyn D.; Sato, Michio; Lian, Mingzhu; Watanabe, Kenji; Rigden, Daniel J.; Kim, Chu-Young (Liverpool); (Daresbury); (NU Singapore); (Shizuoka); (RAL)

    2013-12-02

    Echinomycin is a nonribosomal depsipeptide natural product with a range of interesting bioactivities that make it an important target for drug discovery and development. It contains a thioacetal bridge, a unique chemical motif derived from the disulfide bond of its precursor antibiotic triostin A by the action of an S-adenosyl-L-methionine-dependent methyltransferase, Ecm18. The crystal structure of Ecm18 in complex with its reaction products S-adenosyl-L-homocysteine and echinomycin was determined at 1.50 Å resolution. Phasing was achieved using a new molecular replacement package called AMPLE, which automatically derives search models from structure predictions based on ab initio protein modelling. Structural analysis indicates that a combination of proximity effects, medium effects, and catalysis by strain drives the unique transformation of the disulfide bond into the thioacetal linkage.

  20. reaxFF Reactive Force Field for Disulfide Mechanochemistry, Fitted to Multireference ab Initio Data.

    Science.gov (United States)

    Müller, Julian; Hartke, Bernd

    2016-08-01

    Mechanochemistry, in particular in the form of single-molecule atomic force microscopy experiments, is difficult to model theoretically, for two reasons: Covalent bond breaking is not captured accurately by single-determinant, single-reference quantum chemistry methods, and experimental times of milliseconds or longer are hard to simulate with any approach. Reactive force fields have the potential to alleviate both problems, as demonstrated in this work: Using nondeterministic global parameter optimization by evolutionary algorithms, we have fitted a reaxFF force field to high-level multireference ab initio data for disulfides. The resulting force field can be used to reliably model large, multifunctional mechanochemistry units with disulfide bonds as designed breaking points. Explorative calculations show that a significant part of the time scale gap between AFM experiments and dynamical simulations can be bridged with this approach. PMID:27415976

  1. Batch and Flow Synthesis of Disulfides by Visible-Light-Induced TiO2 Photocatalysis.

    Science.gov (United States)

    Bottecchia, Cecilia; Erdmann, Nico; Tijssen, Patricia M A; Milroy, Lech-Gustav; Brunsveld, Luc; Hessel, Volker; Noël, Timothy

    2016-07-21

    A mild and practical method for the preparation of disulfides through visible-light-induced photocatalytic aerobic oxidation of thiols has been developed. The method involves the use of TiO2 as a heterogeneous photocatalyst. The catalyst's high stability and recyclability makes this method highly practical. The reaction can be substantially accelerated in a continuous-flow packed-bed reactor, which enables a safe and reliable scale-up of the reaction conditions. The batch and flow protocol described herein can be applied to a diverse set of thiol substrates for the preparation of homo- and hetero-dimerized disulfides. Furthermore, biocompatible reaction conditions (i.e., room temperature, visible light, neutral buffer solution, and no additional base) have been developed, which permits the rapid and chemoselective modification of densely functionalized peptide substrates without recourse to complex purification steps. PMID:27329945

  2. Batch and Flow Synthesis of Disulfides by Visible-Light-Induced TiO2 Photocatalysis.

    Science.gov (United States)

    Bottecchia, Cecilia; Erdmann, Nico; Tijssen, Patricia M A; Milroy, Lech-Gustav; Brunsveld, Luc; Hessel, Volker; Noël, Timothy

    2016-07-21

    A mild and practical method for the preparation of disulfides through visible-light-induced photocatalytic aerobic oxidation of thiols has been developed. The method involves the use of TiO2 as a heterogeneous photocatalyst. The catalyst's high stability and recyclability makes this method highly practical. The reaction can be substantially accelerated in a continuous-flow packed-bed reactor, which enables a safe and reliable scale-up of the reaction conditions. The batch and flow protocol described herein can be applied to a diverse set of thiol substrates for the preparation of homo- and hetero-dimerized disulfides. Furthermore, biocompatible reaction conditions (i.e., room temperature, visible light, neutral buffer solution, and no additional base) have been developed, which permits the rapid and chemoselective modification of densely functionalized peptide substrates without recourse to complex purification steps.

  3. Scanning tunneling microscopy on CVD grown lateral graphene molybdenum disulfide heterostructures

    Science.gov (United States)

    Kerelsky, Alexander; Cheng, Minghao; Zhong, Xinjue; Zhao, Xiaodong; Dadgar, Ali; Wang, Da; Gao, Hui; Guimaraes, Marcos; Kang, Kibum; Zhu, Xiaoyang; Park, Jiwoong; Pasupathy, Abhay N.

    We investigate the interface of single layer graphene, molybdenum disulfide lateral heterostructures using scanning tunneling microscopy (STM). Samples are fabricated using chemical vapor deposition to deposit graphene, photolithography to pattern graphene and metal-organic chemical vapor deposition to grow molybdenum disulfide in patterned areas. The lateral junction of the two materials allows investigation of structural and electronic properties at the interface of the two materials, an interface usually buried in conventional stacked heterostructures. STM is used to image the stitching of the two materials with nanoscale resolution. STM is also used to perform local spectroscopy, probing the local density of states on an atomic scale across the junction. Interesting phenomena such as the charge transfer and atomic bonding are investigated. The spatially changing chemical potential between the two materials is also examined at different gate voltages.

  4. Rational Design of a Fusion Protein to Exhibit Disulfide-Mediated Logic Gate Behavior

    Science.gov (United States)

    2015-01-01

    Synthetic cellular logic gates are primarily built from gene circuits owing to their inherent modularity. Single proteins can also possess logic gate functions and offer the potential to be simpler, quicker, and less dependent on cellular resources than gene circuits. However, the design of protein logic gates that are modular and integrate with other cellular components is a considerable challenge. As a step toward addressing this challenge, we describe the design, construction, and characterization of AND, ORN, and YES logic gates built by introducing disulfide bonds into RG13, a fusion of maltose binding protein and TEM-1 β-lactamase for which maltose is an allosteric activator of enzyme activity. We rationally designed these disulfide bonds to manipulate RG13’s allosteric regulation mechanism such that the gating had maltose and reducing agents as input signals, and the gates could be toggled between different gating functions using redox agents, although some gates performed suboptimally. PMID:25144732

  5. Sample Limited Characterization of a Novel Disulfide-Rich Venom Peptide Toxin from Terebrid Marine Snail Terebra variegata

    OpenAIRE

    Anand, Prachi; Grigoryan, Alexandre; Bhuiyan, Mohammed H.; Ueberheide, Beatrix; Russell, Victoria; Quinoñez, Jose; Moy, Patrick; Brian T. Chait; Poget, Sébastien F.; Holford, Mandë

    2014-01-01

    Disulfide-rich peptide toxins found in the secretions of venomous organisms such as snakes, spiders, scorpions, leeches, and marine snails are highly efficient and effective tools for novel therapeutic drug development. Venom peptide toxins have been used extensively to characterize ion channels in the nervous system and platelet aggregation in haemostatic systems. A significant hurdle in characterizing disulfide-rich peptide toxins from venomous animals is obtaining significant quantities ne...

  6. Disulfide Bond in Pseudomonas aeruginosa Lipase Stabilizes the Structure but Is Not Required for Interaction with Its Foldase

    OpenAIRE

    Liebeton, Klaus; Zacharias, Annette; Jaeger, Karl-Erich

    2001-01-01

    Pseudomonas aeruginosa secretes a 29-kDa lipase which is dependent for folding on the presence of the lipase-specific foldase Lif. The lipase contains two cysteine residues which form an intramolecular disulfide bond. Variant lipases with either one or both cysteines replaced by serines showed severely reduced levels of extracellular lipase activity, indicating the importance of the disulfide bond for secretion of lipase through the outer membrane. Wild-type and variant lipase genes fused to ...

  7. Disruption of reducing pathways is not essential for efficient disulfide bond formation in the cytoplasm of E. coli

    Directory of Open Access Journals (Sweden)

    Hatahet Feras

    2010-09-01

    Full Text Available Abstract Background The formation of native disulfide bonds is a complex and essential post-translational modification for many proteins. The large scale production of these proteins can be difficult and depends on targeting the protein to a compartment in which disulfide bond formation naturally occurs, usually the endoplasmic reticulum of eukaryotes or the periplasm of prokaryotes. It is currently thought to be impossible to produce large amounts of disulfide bond containing protein in the cytoplasm of wild-type bacteria such as E. coli due to the presence of multiple pathways for their reduction. Results Here we show that the introduction of Erv1p, a sulfhydryl oxidase and FAD-dependent catalyst of disulfide bond formation found in the inter membrane space of mitochondria, allows the efficient formation of native disulfide bonds in heterologously expressed proteins in the cytoplasm of E. coli even without the disruption of genes involved in disulfide bond reduction, for example trxB and/or gor. Indeed yields of active disulfide bonded proteins were higher in BL21 (DE3 pLysSRARE, an E. coli strain with the reducing pathways intact, than in the commercial Δgor ΔtrxB strain rosetta-gami upon co-expression of Erv1p. Conclusions Our results refute the current paradigm in the field that disruption of at least one of the reducing pathways is essential for the efficient production of disulfide bond containing proteins in the cytoplasm of E. coli and open up new possibilities for the use of E. coli as a microbial cell factory.

  8. Intracellular reduction/activation of a disulfide switch in thiosemicarbazone iron chelators

    OpenAIRE

    Akam, Eman A.; Chang, Tsuhen M.; Astashkin, Andrei V.; Tomat, Elisa

    2014-01-01

    Iron scavengers (chelators) offer therapeutic opportunities in anticancer drug design by targeting the increased demand for iron in cancer cells as compared to normal cells. Prochelation approaches are expected to avoid systemic iron depletion as chelators are liberated under specific intracellular conditions. In the strategy described herein, a disulfide linkage is employed as a redox-directed switch within the binding unit of an antiproliferative thiosemicarbazone prochelator, which is acti...

  9. Control of stability of polypeptide multilayer nanofilms by quantitative control of disulfide bond formation

    International Nuclear Information System (INIS)

    The crosslinking of polymers in a polymeric material will alter the mechanical properties of the material. Control over the mechanical properties of polyelectrolyte multilayer films (PEMs) could be useful for applications of the technology in medicine and other areas. Disulfide bonds are 'natural' polypeptide crosslinks found widely in wild-type proteins. Here, we have designed and synthesized three pairs of oppositely charged 32mer polypeptide to have 0, 4, or 8 cysteine (Cys) residues per molecule, and we have characterized physical properties of the peptides in a PEM context. The average linear density of free thiol in the designed peptides was 0, 0.125, or 0.25 per amino acid residue. The peptides were used to make 10-bilayer PEMs by electrostatic layer-by-layer self-assembly (LBL). Cys was included in the peptides to study specific effects of disulfide bond formation on PEM properties. Features of film assembly have been found to depend on the amino acid sequence, as in protein folding. Following polypeptide self-assembly into multilayer films, Cys residues were disulfide-crosslinked under mild oxidizing conditions. The stability of the crosslinked films at acidic pH has been found to depend on the number of Cys residues per peptide for a given crosslinking procedure. Crosslinked and non-crosslinked films have been analysed by ultraviolet spectroscopy (UVS), ellipsometry, and atomic force microscopy (AFM) to characterize film assembly, surface morphology, and disassembly. A selective etching model of the disassembly process at acidic pH is proposed on the basis of the experimental data. In this model, regions of film in which the disulfide bond density is low are etched at a higher rate than regions where the density is high

  10. Control of stability of polypeptide multilayer nanofilms by quantitative control of disulfide bond formation

    Science.gov (United States)

    Zhong, Yang; Li, Bingyun; Haynie, Donald T.

    2006-12-01

    The crosslinking of polymers in a polymeric material will alter the mechanical properties of the material. Control over the mechanical properties of polyelectrolyte multilayer films (PEMs) could be useful for applications of the technology in medicine and other areas. Disulfide bonds are 'natural' polypeptide crosslinks found widely in wild-type proteins. Here, we have designed and synthesized three pairs of oppositely charged 32mer polypeptide to have 0, 4, or 8 cysteine (Cys) residues per molecule, and we have characterized physical properties of the peptides in a PEM context. The average linear density of free thiol in the designed peptides was 0, 0.125, or 0.25 per amino acid residue. The peptides were used to make 10-bilayer PEMs by electrostatic layer-by-layer self-assembly (LBL). Cys was included in the peptides to study specific effects of disulfide bond formation on PEM properties. Features of film assembly have been found to depend on the amino acid sequence, as in protein folding. Following polypeptide self-assembly into multilayer films, Cys residues were disulfide-crosslinked under mild oxidizing conditions. The stability of the crosslinked films at acidic pH has been found to depend on the number of Cys residues per peptide for a given crosslinking procedure. Crosslinked and non-crosslinked films have been analysed by ultraviolet spectroscopy (UVS), ellipsometry, and atomic force microscopy (AFM) to characterize film assembly, surface morphology, and disassembly. A selective etching model of the disassembly process at acidic pH is proposed on the basis of the experimental data. In this model, regions of film in which the disulfide bond density is low are etched at a higher rate than regions where the density is high.

  11. Pantethine and cystamine deplete cystine from cystinotic fibroblasts via efflux of cysteamine-cysteine mixed disulfide.

    OpenAIRE

    Butler, J D; Zatz, M

    1984-01-01

    Children suffering from cystinosis, a genetic disease characterized by high levels of lysosomal cystine, are currently being treated with cysteamine to lower the cystine levels in their cells. In fibroblasts from these patients, cysteamine and its disulfide, cystamine, are equally effective in lowering cystine levels. We recently reported that pantethine, a dietary precursor of coenzyme A, depletes cystine from cultured, cystinotic fibroblasts as effectively as cystamine. To determine the mec...

  12. The drosomycin multigene family: three-disulfide variants from Drosophila takahashii possess antibacterial activity.

    Science.gov (United States)

    Gao, Bin; Zhu, Shunyi

    2016-01-01

    Drosomycin (DRS) is a strictly antifungal peptide in Drosophila melanogaster, which contains four disulfide bridges (DBs) with three buried in molecular interior and one exposed on molecular surface to tie the amino- and carboxyl-termini of the molecule together (called wrapper disulfide bridge, WDB). Based on computational analysis of genomes of Drosophila species belonging to the Oriental lineage, we identified a new multigene family of DRS in Drosphila takahashii that includes a total of 11 DRS-encoding genes (termed DtDRS-1 to DtDRS-11) and a pseudogene. Phylogenetic tree and synteny analyses reveal orthologous relationship between DtDRSs and DRSs, indicating that orthologous genes of DRS-1, DRS-2, DRS-3 and DRS-6 have undergone duplication in D. takahashii and three amplifications (DtDRS-9 to DtDRS-11) of DRS-3 have lost WDB. Among the 11 genes, five are transcriptionally active in adult fruitflies. The ortholog of DRS (DtDRS-1) shows high structural and functional similarity to DRS while two WDB-deficient members display antibacterial activity accompanying complete loss or remarkable reduction of antifungal activity. To the best of our knowledge, this is the first report on the presence of three-disulfide antibacterial DRSs in a specific Drosophila species, suggesting a potential role of DB loss in neofunctionalization of a protein via structural adjustment. PMID:27562645

  13. Resolution of Disulfide Heterogeneity in Nogo Receptor 1 Fusion Proteins by Molecular Engineering

    Energy Technology Data Exchange (ETDEWEB)

    P Weinreb; D Wen; F Qian; C Wildes; E Garber; L Walus; M Jung; J Wang; J Relton; et al.

    2011-12-31

    NgRI (Nogo-66 receptor) is part of a signalling complex that inhibits axon regeneration in the central nervous system. Truncated soluble versions of NgRI have been used successfully to promote axon regeneration in animal models of spinal-cord injury, raising interest in this protein as a potential therapeutic target. The LRR (leucine-rich repeat) regions in NgRI are flanked by N- and C-terminal disulfide-containing 'cap' domains (LRRNT and LRRCT respectively). In the present work we show that, although functionally active, the NgRI(310)-Fc fusion protein contains mislinked and heterogeneous disulfide patterns in the LRRCT domain, and we report the generation of a series of variant molecules specifically designed to prevent this heterogeneity. Using these variants we explored the effects of modifying the NgRI truncation site or the spacing between the NgRI and Fc domains, or replacing cysteines within the NgRI or IgG hinge regions. One variant, which incorporates replacements of Cys{sup 266} and Cys{sup 309} with alanine residues, completely eliminated disulfide scrambling while maintaining functional in vitro and in vivo efficacy. This modified NgRI-Fc molecule represents a significantly improved candidate for further pharmaceutical development, and may serve as a useful model for the optimization of other IgG fusion proteins made from LRR proteins.

  14. Poly(vinyl alcohol) nanocomposite films containing chemically exfoliated molybdenum disulfide

    Energy Technology Data Exchange (ETDEWEB)

    Batista Mancinelli, Ketlin Cristine; Lisboa, Fabio da Silva [Centro de Pesquisa em Quimica Aplicada - CEPESQ, Laboratorio de Quimica de Materiais Avancados, Universidade Federal do Parana, Departamento de Quimica, Caixa Postal 19081, 81531-980 Curitiba, PR (Brazil); Soares, Jaisa Fernandes [Laboratorio de Quimica Bioinorganica, Universidade Federal do Parana, Departamento de Quimica, Caixa Postal 19081, 81531-980 Curitiba, PR (Brazil); Zawadzki, Sonia Faria [Laboratorio de Polimeros Sinteticos, Universidade Federal do Parana, Departamento de Quimica, Caixa Postal 19081, 81531-980 Curitiba, PR (Brazil); Wypych, Fernando, E-mail: wypych@ufpr.br [Centro de Pesquisa em Quimica Aplicada - CEPESQ, Laboratorio de Quimica de Materiais Avancados, Universidade Federal do Parana, Departamento de Quimica, Caixa Postal 19081, 81531-980 Curitiba, PR (Brazil)

    2013-01-15

    Molybdenum disulfide (2H-MoS{sub 2}) was exfoliated in water after reaction with n-butyl-lithium. Using either alkaline or neutral conditions, different amounts of the resulting single-layer suspension were employed as filler for the production of poly(vinyl alcohol) films containing distinct disulfide contents. These nanocomposite films were obtained by wet casting and were further characterized by powder X-ray diffraction (XRD), differential scanning calorimetry (DSC), and Fourier transform infrared (FTIR) and Raman spectroscopies. The mechanical properties of the films were also evaluated. Characterization studies revealed the attainment of homogeneous nanocomposite films in both alkaline and neutral conditions, indicating good distribution and interaction of the hydrophilic filler with the polyhydroxylated polymer. Improved Young's (tensile) modulus (+57%) and tensile strength (+9%) as well as reduced elongation (-78%) were achieved only when the neutral suspension of single layers was utilized. Increased MoS{sub 2} content diminished the crystallinity of the polymer, while enhanced mechanical properties were obtained in the presence of intermediate filler content (around 1 wt%). Highlights: Black-Right-Pointing-Pointer Molybdenum disulfide (2H-MoS{sub 2}) was chemically exfoliated in water. Black-Right-Pointing-Pointer MoS{sub 2} single-layer suspension was used as filler for poly(vinyl alcohol) films. Black-Right-Pointing-Pointer Increased MoS{sub 2} content diminished the crystallinity of the polymer. Black-Right-Pointing-Pointer Enhanced mechanical properties were obtained with intermediate filler content.

  15. Site-directed introduction of disulfide groups on antibodies for highly sensitive immunosensors.

    Science.gov (United States)

    Acero Sánchez, Josep Ll; Fragoso, Alex; Joda, Hamdi; Suárez, Guillaume; McNeil, Calum J; O'Sullivan, Ciara K

    2016-07-01

    The interface between the sample and the transducer surface is critical to the performance of a biosensor. In this work, we compared different strategies for covalent self-assembly of antibodies onto bare gold substrates by introducing disulfide groups into the immunoglobulin structure, which acted as anchor molecules able to chemisorb spontaneously onto clean gold surfaces. The disulfide moieties were chemically introduced to the antibody via the primary amines, carboxylic acids, and carbohydrates present in its structure. The site-directed modification via the carbohydrate chains exhibited the best performance in terms of analyte response using a model system for the detection of the stroke marker neuron-specific enolase. SPR measurements clearly showed the potential for creating biologically active densely packed self-assembled monolayers (SAMs) in a one-step protocol compared to both mixed SAMs of alkanethiol compounds and commercial immobilization layers. The ability of the carbohydrate strategy to construct an electrochemical immunosensor was investigated using electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) transduction. Graphical Abstract Left: Functionalization strategies of bare gold substrates via direct bio-SAM using disulfide-containing antibody chemically modified via their primary amines (A), carbohydrates (B) and carboxylic acids (C). Right: Dependence of the peak height with NSE concentration at NSE21-CHO modified electrochemical immunosensor. Inset: Logarithmic calibration plot. PMID:27220524

  16. Oxidative modification of caspase-9 facilitates its activation via disulfide-mediated interaction with Apaf-1

    Institute of Scientific and Technical Information of China (English)

    Yong Zuo; Binggang Xiang; Jie Yang; Xuxu Sun; Yumei Wang; Hui Cang; Jing Yi

    2009-01-01

    Intracellular reactive oxygen species (ROS) are known to regulate apoptosis. Activation of caspase-9, the initial caspase in the mitochondrial apoptotic cascade, is closely associated with ROS, but it is unclear whether ROS regulate caspase-9 via direct oxidative modification. The present study aims to elucidate the molecular mechanisms by which ROS mediate caspase-9 activation. Our results show that the cellular oxidative state facilitates caspase-9 activation. Hydrogen peroxide treatment causes the activation of caspase-9 and apoptosis, and promotes an interaction between caspase-9 and apoptotic protease-activating factor 1 (Apaf-1) via disulfide formation. In addition, in an in vitro mitochondria-free system, the thiol-oxidant diamide promotes auto-cleavage of caspase-9 and the caspase-9/ Apaf-1 interaction by facilitating the formation of disulfide-linked complexes. Finally, a point mutation at C403 of caspase-9 impairs both H202-promoted caspase-9 activation and interaction with Apaf-1 through the abolition of disulfide formation. The association between cytochrome c and the C403S mutant is significantly weaker than that between cytochrome c and wild-type caspase-9, indicating that oxidative modification of caspase-9 contributes to apoptosome formation under oxidative stress. Taken together, oxidative modification of caspase-9 by ROS can mediate its interaction with Apaf-1, and can thus promote its auto-cleavage and activation. This mechanism may facilitate apoptosome formation and caspase-9 activation under oxidative stress.

  17. Biocompatible thermoresponsive PEGMA nanoparticles crosslinked with cleavable disulfide-based crosslinker for dual drug release.

    Science.gov (United States)

    Ulasan, Mehmet; Yavuz, Emine; Bagriacik, Emin Umit; Cengeloglu, Yunus; Yavuz, Mustafa Selman

    2015-01-01

    Smart materials have been attracting much attention because of their stimuli responsive nature. We have synthesized biocompatible thermoresponsive crosslinked poly(ethylene glycol) methyl ether methacrylate (PEGMA)-co-vinyl pyrrolidone nanoparticles (PEGMA NPs) using disulfide-based crosslinker by surfactant-free emulsion polymerization method. Particle characterization studies were carried out by dynamic light scattering, and scanning electron microscopy. Polymerization kinetics, effect of crosslinker and initiator concentrations on both average hydrodynamic diameter and polydispersity index were investigated. Hydrodynamic diameters of thermoresponsive PEGMA NPs were decreased from 210 nm to 90 nm upon heating over the lowest critical solution temperature (LCST). Disulfide crosslinked PEGMA NPs were demonstrated as a dual delivery system. Rhodamine B, a model of small-sized drug molecule, and poly(ethylene glycol) (PEG)-alizarin yellow, a model of large drug molecule, were loaded into PEGMA NPs where LCST of these NPs was tuned to 37°C, the body temperature. The rhodamine B was released from PEGMA NPs upon heating to 39°C. Then, PEG-alizarin content was released by subsequent degradation of nanoparticles using dithiothreitol (DTT), which reduces disulfide bonds to thiols. Furthermore, cytotoxicity studies of PEGMA NPs were carried out in 3T3 cells, which resulted in no toxic effect on the cells.

  18. Model building of disulfide bonds in proteins with known three-dimensional structure.

    Science.gov (United States)

    Hazes, B; Dijkstra, B W

    1988-07-01

    As an aid in the selection of sites in a protein where a disulfide bond might be engineered, a computer program has been developed. The algorithm starts with the generation of C beta positions from the N, C alpha and C atom coordinates available from a three-dimensional model. A first set of residue pairs that might form a disulfide bond is selected on the basis of C beta-C beta distances between residues. Then, for each residue in this set, S gamma positions are generated, which satisfy the requirement that, with ideal values for the C alpha-C beta and C beta-S gamma bond lengths and for the bond angle at C beta, the distance between S gamma of residue 1 and C beta of residue 2 in a pair (determined by the bond angle at S gamma 2) is at, or very close to its ideal value. Usually two acceptable S gamma positions are found for each half cystine, resulting in up to four different conformations for the disulfide bond. Finally, these conformations are subjected to an energy minimization procedure to remove large deviations from ideal geometry and their final energies are calculated. User input determines which final conformations are energetically acceptable. These conformations are written to a file to allow further analysis and e.g. inspection on a computer graphics device. PMID:3244694

  19. Protein disulfide-isomerase interacts with a substrate protein at all stages along its folding pathway.

    Directory of Open Access Journals (Sweden)

    Alistair G Irvine

    Full Text Available In contrast to molecular chaperones that couple protein folding to ATP hydrolysis, protein disulfide-isomerase (PDI catalyzes protein folding coupled to formation of disulfide bonds (oxidative folding. However, we do not know how PDI distinguishes folded, partly-folded and unfolded protein substrates. As a model intermediate in an oxidative folding pathway, we prepared a two-disulfide mutant of basic pancreatic trypsin inhibitor (BPTI and showed by NMR that it is partly-folded and highly dynamic. NMR studies show that it binds to PDI at the same site that binds peptide ligands, with rapid binding and dissociation kinetics; surface plasmon resonance shows its interaction with PDI has a Kd of ca. 10(-5 M. For comparison, we characterized the interactions of PDI with native BPTI and fully-unfolded BPTI. Interestingly, PDI does bind native BPTI, but binding is quantitatively weaker than with partly-folded and unfolded BPTI. Hence PDI recognizes and binds substrates via permanently or transiently unfolded regions. This is the first study of PDI's interaction with a partly-folded protein, and the first to analyze this folding catalyst's changing interactions with substrates along an oxidative folding pathway. We have identified key features that make PDI an effective catalyst of oxidative protein folding - differential affinity, rapid ligand exchange and conformational flexibility.

  20. Characterization of Sviceucin from Streptomyces Provides Insight into Enzyme Exchangeability and Disulfide Bond Formation in Lasso Peptides.

    Science.gov (United States)

    Li, Yanyan; Ducasse, Rémi; Zirah, Séverine; Blond, Alain; Goulard, Christophe; Lescop, Ewen; Giraud, Caroline; Hartke, Axel; Guittet, Eric; Pernodet, Jean-Luc; Rebuffat, Sylvie

    2015-11-20

    Lasso peptides are bacterial ribosomally synthesized and post-translationally modified peptides. They have sparked increasing interest in peptide-based drug development because of their compact, interlocked structure, which offers superior stability and protein-binding capacity. Disulfide bond-containing lasso peptides are rare and exhibit highly sought-after activities. In an effort to expand the repertoire of such molecules, we heterologously expressed, in Streptomyces coelicolor, the gene cluster encoding sviceucin, a type I lasso peptide with two disulfide bridges originating from Streptomyces sviceus, which allowed it to be fully characterized. Sviceucin and its reduced forms were characterized by mass spectrometry and peptidase digestion. The three-dimensional structure of sviceucin was determined using NMR. Sviceucin displayed antimicrobial activity selectively against Gram-positive bacteria and inhibition of fsr quorum sensing in Enterococcus faecalis. This study adds sviceucin to the type I lasso peptide family as a new representative. Moreover, new clusters encoding disulfide-bond containing lasso peptides from Actinobacteria were identified by genome mining. Genetic and functional analyses revealed that the formation of disulfide bonds in sviceucin does not require a pathway-encoded thiol-disulfide oxidoreductase. Most importantly, we demonstrated the functional exchangeability of the sviceucin and microcin J25 (a non-disulfide-bridged lasso peptide) macrolactam synthetases in vitro, highlighting the potential of hybrid lasso synthetases in lasso peptide engineering. PMID:26343290

  1. Ion Mobility-Mass Spectrometry as a Tool for the Structural Characterization of Peptides Bearing Intramolecular Disulfide Bond(s)

    Science.gov (United States)

    Massonnet, Philippe; Haler, Jean R. N.; Upert, Gregory; Degueldre, Michel; Morsa, Denis; Smargiasso, Nicolas; Mourier, Gilles; Gilles, Nicolas; Quinton, Loïc; De Pauw, Edwin

    2016-10-01

    Disulfide bonds are post-translationnal modifications that can be crucial for the stability and the biological activities of natural peptides. Considering the importance of these disulfide bond-containing peptides, the development of new techniques in order to characterize these modifications is of great interest. For this purpose, collision cross cections (CCS) of a large data set of 118 peptides (displaying various sequences) bearing zero, one, two, or three disulfide bond(s) have been measured in this study at different charge states using ion mobility-mass spectrometry. From an experimental point of view, CCS differences (ΔCCS) between peptides bearing various numbers of disulfide bonds and peptides having no disulfide bonds have been calculated. The ΔCCS calculations have also been applied to peptides bearing two disulfide bonds but different cysteine connectivities (Cys1-Cys2/Cys3-Cys4; Cys1-Cys3/Cys2-Cys4; Cys1-Cys4/Cys2-Cys3). The effect of the replacement of a proton by a potassium adduct on a peptidic structure has also been investigated.

  2. Thiol/Disulfide system plays a crucial role in redox protection in the acidophilic iron-oxidizing bacterium Leptospirillum ferriphilum.

    Directory of Open Access Journals (Sweden)

    Javiera Norambuena

    Full Text Available Thiol/disulfide systems are involved in the maintenance of the redox status of proteins and other molecules that contain thiol/disulfide groups. Leptospirillum ferriphilum DSM14647, an acidophilic bacterium that uses Fe(2+ as electron donor, and withstands very high concentrations of iron and other redox active metals, is a good model to study how acidophiles preserve the thiol/disulfide balance. We studied the composition of thiol/disulfide systems and their role in the oxidative stress response in this extremophile bacterium. Bioinformatic analysis using genomic data and enzymatic assays using protein extracts from cells grown under oxidative stress revealed that the major thiol/disulfide system from L. ferriphilum are a cytoplasmic thioredoxin system (composed by thioredoxins Trx and thioredoxin reductase TR, periplasmic thiol oxidation system (DsbA/DsbB and a c-type cytochrome maturation system (DsbD/DsbE. Upon exposure of L. ferriphilum to reactive oxygen species (ROS-generating compounds, transcriptional activation of the genes encoding Trxs and the TR enzyme, which results in an increase of the corresponding activity, was observed. Altogether these data suggest that the thioredoxin-based thiol/disulfide system plays an important role in redox protection of L. ferriphilum favoring the survival of this microorganism under extreme environmental oxidative conditions.

  3. Ion Mobility-Mass Spectrometry as a Tool for the Structural Characterization of Peptides Bearing Intramolecular Disulfide Bond(s)

    Science.gov (United States)

    Massonnet, Philippe; Haler, Jean R. N.; Upert, Gregory; Degueldre, Michel; Morsa, Denis; Smargiasso, Nicolas; Mourier, Gilles; Gilles, Nicolas; Quinton, Loïc; De Pauw, Edwin

    2016-08-01

    Disulfide bonds are post-translationnal modifications that can be crucial for the stability and the biological activities of natural peptides. Considering the importance of these disulfide bond-containing peptides, the development of new techniques in order to characterize these modifications is of great interest. For this purpose, collision cross cections (CCS) of a large data set of 118 peptides (displaying various sequences) bearing zero, one, two, or three disulfide bond(s) have been measured in this study at different charge states using ion mobility-mass spectrometry. From an experimental point of view, CCS differences (ΔCCS) between peptides bearing various numbers of disulfide bonds and peptides having no disulfide bonds have been calculated. The ΔCCS calculations have also been applied to peptides bearing two disulfide bonds but different cysteine connectivities (Cys1-Cys2/Cys3-Cys4; Cys1-Cys3/Cys2-Cys4; Cys1-Cys4/Cys2-Cys3). The effect of the replacement of a proton by a potassium adduct on a peptidic structure has also been investigated.

  4. Secondary Ion Mass Spectrometry of Small-Molecule Solids at Cryogenic Temperatures. IV. Carbon Dioxide, Carbonyl Sulfide and Carbon Disulfide

    NARCIS (Netherlands)

    Orth, Robert G.; Jonkman, Harry T.; Michl, Josef

    1982-01-01

    Secondary ion mass spectra of neat solid CO2, COS and CS2 and of CO2 diluted in solid argon were measured as a function of the nature and energy of the primary ions (He+, Ne+, Ar+, Kr+, Xe+, 1.0-4.5 keV). All of the solids produced a rich variety of positive and negative secondary ions. Many of thes

  5. Synergistic neurotoxicity of carbon tetrachloride/carbon disulfide (80/20 fumigants) and other pesticides in grain storage workers.

    Science.gov (United States)

    Peters, H A; Levine, R L; Matthews, C G; Sauter, S; Chapman, L

    1986-01-01

    Neurophysiologic, neurobehavioral, and neuropsychologic profiles in 17 grain storage workers, 1 grain inspector, and 4 malting laboratory workers are described. The effects of CS2 toxicity as seen in viscose rayon workers as well as in experimental animals is remarkably similar to the clinical profile of our grain storage workers. CS2 use explains the dysfunction of peripheral axons, auditory nerve, the optic nerve, and the extrapyramidal system, as well as altered behavior and cognition changes. The signs and symptoms in these workers seem to be dose-related and we note that workers separated out from the areas where fumigation took place reported improvement not seen by fellow workers who continued the fumigant treatment routine. Likewise, malting laboratory workers exposed only to the grain dust from 3 to 7 years showed only minimal symptoms. Though a number of mechanism have been suggested for the alteration of neuropsychological function, the chelating ability of DDC derived from CS2 and its ability to markedly increase copper and zinc within the central nervous system suggests a mechanism of toxicity analogous to copper intoxication as in Wilson's Disease and may explain the production of extrapyramidal symptoms in these patients. Chelation of copper might prove therapeutic in CS2 poisoning. It is obvious that both basic and clinical research will be necessary to sort out the questions raised. We applaud the EPA's decision to ban the use of 80/20 fumigants and also methyl bromide, and trust that similar toxic substances be carefully studied before their selection for replacing these previous toxic agents. We further decry the technique of re-introducing grain dust into the food chain rather than destroying it, since the dust contains very high residues of fumigant material. We speculate on the possible role of CS2 and other pesticides in the food chain and the incidence of Parkinsonian symptoms in these patients and the general public.

  6. Dissecting a Role of Evolutionary-conserved but Non-critical Disulfide Bridges in Cysteine-Rich Peptides Using ω-Conotoxin GVIA and its Selenocysteine Analogs

    OpenAIRE

    Gowd, Konkallu Hanumae; Blais, Kirk D.; Elmslie, Keith S.; Andrew M Steiner; Olivera, Baldomero M.; Bulaj, Grzegorz

    2012-01-01

    Conotoxins comprise a large group of peptidic neurotoxins that employ diverse disulfide-rich scaffolds. Each scaffold is determined by an evolutionarily conserved pattern of cysteine residues. Although many structure-activity relationship studies confirm the functional and structural importance of disulfide crosslinks, there is growing evidence that not all disulfide bridges are critical in maintaining activities of conotoxins. To answer the fundamental biological question of what the role of...

  7. Structural studies of polypeptides: Mechanism of immunoglobin catalysis and helix propagation in hybrid sequence, disulfide containing peptides

    Energy Technology Data Exchange (ETDEWEB)

    Storrs, R.W.

    1992-08-01

    Catalytic immunoglobin fragments were studied Nuclear Magnetic Resonance spectroscopy to identify amino acid residues responsible for the catalytic activity. Small, hybrid sequence peptides were analyzed for helix propagation following covalent initiation and for activity related to the protein from which the helical sequence was derived. Hydrolysis of p-nitrophenyl carbonates and esters by specific immunoglobins is thought to involve charge complementarity. The pK of the transition state analog P-nitrophenyl phosphate bound to the immunoglobin fragment was determined by {sup 31}P-NMR to verify the juxtaposition of a positively charged amino acid to the binding/catalytic site. Optical studies of immunoglobin mediated photoreversal of cis, syn cyclobutane thymine dimers implicated tryptophan as the photosensitizing chromophore. Research shows the chemical environment of a single tryptophan residue is altered upon binding of the thymine dimer. This tryptophan residue was localized to within 20 {Angstrom} of the binding site through the use of a nitroxide paramagnetic species covalently attached to the thymine dimer. A hybrid sequence peptide was synthesized based on the bee venom peptide apamin in which the helical residues of apamin were replaced with those from the recognition helix of the bacteriophage 434 repressor protein. Oxidation of the disufide bonds occured uniformly in the proper 1-11, 3-15 orientation, stabilizing the 434 sequence in an {alpha}-helix. The glycine residue stopped helix propagation. Helix propagation in 2,2,2-trifluoroethanol mixtures was investigated in a second hybrid sequence peptide using the apamin-derived disulfide scaffold and the S-peptide sequence. The helix-stop signal previously observed was not observed in the NMR NOESY spectrum. Helical connectivities were seen throughout the S-peptide sequence. The apamin/S-peptide hybrid binded to the S-protein (residues 21-166 of ribonuclease A) and reconstituted enzymatic activity.

  8. Structural studies of polypeptides: Mechanism of immunoglobin catalysis and helix propagation in hybrid sequence, disulfide containing peptides

    Energy Technology Data Exchange (ETDEWEB)

    Storrs, R.W.

    1992-08-01

    Catalytic immunoglobin fragments were studied Nuclear Magnetic Resonance spectroscopy to identify amino acid residues responsible for the catalytic activity. Small, hybrid sequence peptides were analyzed for helix propagation following covalent initiation and for activity related to the protein from which the helical sequence was derived. Hydrolysis of p-nitrophenyl carbonates and esters by specific immunoglobins is thought to involve charge complementarity. The pK of the transition state analog P-nitrophenyl phosphate bound to the immunoglobin fragment was determined by [sup 31]P-NMR to verify the juxtaposition of a positively charged amino acid to the binding/catalytic site. Optical studies of immunoglobin mediated photoreversal of cis, syn cyclobutane thymine dimers implicated tryptophan as the photosensitizing chromophore. Research shows the chemical environment of a single tryptophan residue is altered upon binding of the thymine dimer. This tryptophan residue was localized to within 20 [Angstrom] of the binding site through the use of a nitroxide paramagnetic species covalently attached to the thymine dimer. A hybrid sequence peptide was synthesized based on the bee venom peptide apamin in which the helical residues of apamin were replaced with those from the recognition helix of the bacteriophage 434 repressor protein. Oxidation of the disufide bonds occured uniformly in the proper 1-11, 3-15 orientation, stabilizing the 434 sequence in an [alpha]-helix. The glycine residue stopped helix propagation. Helix propagation in 2,2,2-trifluoroethanol mixtures was investigated in a second hybrid sequence peptide using the apamin-derived disulfide scaffold and the S-peptide sequence. The helix-stop signal previously observed was not observed in the NMR NOESY spectrum. Helical connectivities were seen throughout the S-peptide sequence. The apamin/S-peptide hybrid binded to the S-protein (residues 21-166 of ribonuclease A) and reconstituted enzymatic activity.

  9. Influence of Disulfide Connectivity on Structure and Bioactivity of α-Conotoxin TxIA

    Directory of Open Access Journals (Sweden)

    Yong Wu

    2014-01-01

    Full Text Available Cone snails express a sophisticated arsenal of small bioactive peptides known as conopeptides or conotoxins (CTxs. Through evolutionary selection, these peptides have gained the ability to interact with a range of ion channels and receptors, such as nicotinic acetylcholine receptors (nAChRs. Here, we used reversed-phase high performance liquid chromatography (RP-HPLC and electrospray ionization-mass spectrometry (ESI-MS to explore the venom peptide diversity of Conus textile, a species of cone snail native to Hainan, China. One fraction of C. textile crude venom potently blocked α3β2 nAChRs. Subsequent purification, synthesis, and tandem mass spectrometric analysis demonstrated that the most active compound in this fraction was identical to α-CTx TxIA, an antagonist of α3β2 nAChRs. Then three disulfide isoforms of α-CTx TxIA were synthesized and their activities were investigated systematically for the first time. As we observed, disulfide isomerisation was particularly important for α-CTx TxIA potency. Although both globular and ribbon isomers showed similar retention times in RP-HPLC, globular TxIA potently inhibited α3β2 nAChRs with an IC50 of 5.4 nM, while ribbon TxIA had an IC50 of 430 nM. In contrast, beads isomer had little activity towards α3β2 nAChRs. Two-step oxidation synthesis produced the highest yield of α-CTx TxIA native globular isomer, while a one-step production process based on random oxidation folding was not suitable. In summary, this study demonstrated the relationship between conotoxin activity and disulfide connectivity on α-CTx TxIA.

  10. An intersubunit disulfide bridge stabilizes the tetrameric nucleoside diphosphate kinase of Aquifex aeolicus.

    Science.gov (United States)

    Boissier, Fanny; Georgescauld, Florian; Moynié, Lucile; Dupuy, Jean-William; Sarger, Claude; Podar, Mircea; Lascu, Ioan; Giraud, Marie-France; Dautant, Alain

    2012-06-01

    The nucleoside diphosphate kinase (Ndk) catalyzes the reversible transfer of the γ-phosphate from nucleoside triphosphate to nucleoside diphosphate. Ndks form hexamers or two types of tetramers made of the same building block, namely, the common dimer. The secondary interfaces of the Type I tetramer found in Myxococcus xanthus Ndk and of the Type II found in Escherichia coli Ndk involve the opposite sides of subunits. Up to now, the few available structures of Ndk from thermophiles were hexameric. Here, we determined the X-ray structures of four crystal forms of the Ndk from the hyperthermophilic bacterium Aquifex aeolicus (Aa-Ndk). Aa-Ndk displays numerous features of thermostable proteins and is made of the common dimer but it is a tetramer of Type I. Indeed, the insertion of three residues in a surface-exposed spiral loop, named the Kpn-loop, leads to the formation of a two-turn α-helix that prevents both hexamer and Type II tetramer assembly. Moreover, the side chain of the cysteine at position 133, which is not present in other Ndk sequences, adopts two alternate conformations. Through the secondary interface, each one forms a disulfide bridge with the equivalent Cys133 from the neighboring subunit. This disulfide bridge was progressively broken during X-ray data collection by radiation damage. Such crosslinks counterbalance the weakness of the common-dimer interface. A 40% decrease of the kinase activity at 60°C after reduction and alkylation of the protein corroborates the structural relevance of the disulfide bridge on the tetramer assembly and enzymatic function. PMID:22467275

  11. Enhanced gene delivery by chitosan-disulfide-conjugated LMW-PEI for facilitating osteogenic differentiation.

    Science.gov (United States)

    Zhao, Xiaoli; Li, Zhaoyang; Pan, Haobo; Liu, Wenguang; Lv, Minmin; Leung, Frankie; Lu, William W

    2013-05-01

    Chitosan-disulfide-conjugated LMW-PEI (CS-ss-PEI) was designed to combine the biocompatibility of chitosan and the gene delivery ability of polyethylenimine (PEI) using bio-reducible disulfide for bone morphogenetic protein (BMP2) gene delivery in mediating osteogenic differentiation. It was prepared by conjugating low molecular weight PEI (LMW-PEI) to chitosan through oxidization of thiols introduced for the formation of disulfide linkage. The structure, molecular weight and buffer capacity were characterized by Fourier transform infrared (FTIR), light scattering and acid-base titration, respectively. The reduction in molecular weight of CS-ss-PEI by the reducing agent indicated its bio-reducible property. With the increment in the LMW-PEI component, the copolymer showed increased DNA binding ability and formed denser nanocomplexes. CS-ss-PEI exhibited low cytotoxicity in COS-1, HepG2 and 293T cells over the different weight ratios. The transfection efficiency of CS-ss-PEI4 was significantly higher than that of PEI 25k and comparable with Lipofectamine in mediating luciferase expression. Its application for BMP2 gene delivery was confirmed in C2C12 cells by BMP2 expression. For inducing in vitro osteogenic differentiation, CS-ss-PEI4 mediated BMP2 gene delivery showed a stronger effect in MG-63 osteoblast cells and stem cells in terms of alkaline phosphatase activity and mineralization compared with PEI25k and Lipofectamine. This study provides a potential gene delivery system for orthopedic-related disease. PMID:23395816

  12. Differential regulation of tissue thiol-disulfide redox status in a murine model of peritonitis

    Directory of Open Access Journals (Sweden)

    Benton Shana M

    2012-10-01

    Full Text Available Abstract Background Glutathione (GSH/glutathione disulfide (GSSG and cysteine (Cys/cystine (CySS are major redox pools with important roles in cytoprotection. We determined the impact of septic peritonitis on thiol-disulfide redox status in mice. Methods FVB/N mice (6–12 week old; 8/group underwent laparotomy with cecal ligation and puncture (CLP or laparotomy alone (control. Sections of ileum, colon, lung and liver were obtained and GSH, GSSG, Cys and CySS concentrations determined by HPLC 24 h after laparotomy. Redox potential [Eh in millivolts (mV] of the GSH/GSSG and Cys/CySS pools was calculated using the Nernst equation. Data were analyzed by ANOVA (mean ± SE. Results GSH/GSSG Eh in ileum, colon, and liver was significantly oxidized in septic mice versus control mice (ileum: septic −202±4 versus control −228±2 mV; colon: -195±8 versus −214±1 mV; and liver: -194±3 vs. -210±1 mV, all Ph was unchanged with CLP, while liver and lung Cys/CySS Eh became significantly more reducing (liver: septic = −103±3 versus control −90±2 mV; lung: -101±5 versus −81±1 mV, each P Conclusions Septic peritonitis induced by CLP oxidizes ileal and colonic GSH/GSSG redox but Cys/CySS Eh remains unchanged in these intestinal tissues. In liver, CLP oxidizes the GSH/GSSG redox pool and CyS/CySS Eh becomes more reducing; in lung, CLP does not alter GSH/GSSG Eh, and Cys/CySS Eh is less oxidized. CLP-induced infection/inflammation differentially regulates major thiol-disulfide redox pools in this murine model.

  13. Molecular characterization and expression profiling of the protein disulfide isomerase gene family in Brachypodium distachyon L.

    Directory of Open Access Journals (Sweden)

    Chong Zhu

    Full Text Available Protein disulfide isomerases (PDI are involved in catalyzing protein disulfide bonding and isomerization in the endoplasmic reticulum and functions as a chaperone to inhibit the aggregation of misfolded proteins. Brachypodium distachyon is a widely used model plant for temperate grass species such as wheat and barley. In this work, we report the first molecular characterization, phylogenies, and expression profiles of PDI and PDI-like (PDIL genes in B. distachyon in different tissues under various abiotic stresses. Eleven PDI and PDIL genes in the B. distachyon genome by in silico identification were evenly distributed across all five chromosomes. The plant PDI family has three conserved motifs that are involved in catalyzing protein disulfide bonding and isomerization, but a different exon/intron structural organization showed a high degree of structural differentiation. Two pairs of genes (BdPDIL4-1 and BdPDIL4-2; BdPDIL7-1 and BdPDIL7-2 contained segmental duplications, indicating each pair originated from one progenitor. Promoter analysis showed that Brachypodium PDI family members contained important cis-acting regulatory elements involved in seed storage protein synthesis and diverse stress response. All Brachypodium PDI genes investigated were ubiquitously expressed in different organs, but differentiation in expression levels among different genes and organs was clear. BdPDIL1-1 and BdPDIL5-1 were expressed abundantly in developing grains, suggesting that they have important roles in synthesis and accumulation of seed storage proteins. Diverse treatments (drought, salt, ABA, and H2O2 induced up- and down-regulated expression of Brachypodium PDI genes in seedling leaves. Interestingly, BdPDIL1-1 displayed significantly up-regulated expression following all abiotic stress treatments, indicating that it could be involved in multiple stress responses. Our results provide new insights into the structural and functional characteristics of the

  14. Antagonistic effect of disulfide-rich peptide aptamers selected by cDNA display on interleukin-6-dependent cell proliferation

    Energy Technology Data Exchange (ETDEWEB)

    Nemoto, Naoto, E-mail: nemoto@fms.saitama-u.ac.jp [Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan); Innovation Center for Startups, National Institute of Advanced Industrial Science and Technology, 2-2-2 Marunouchi, Chiyoda-ku, Tokyo 100-0005 (Japan); Janusys Corporation, 508, Saitama Industrial Technology Center, Skip City, 3-12-18 Kami-Aoki, Kawaguchi, Saitama 333-0844 (Japan); Tsutsui, Chihiro [Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan); Yamaguchi, Junichi [Innovation Center for Startups, National Institute of Advanced Industrial Science and Technology, 2-2-2 Marunouchi, Chiyoda-ku, Tokyo 100-0005 (Japan); Applied Gene Technology, Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566 (Japan); Ueno, Shingo [Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan); Machida, Masayuki [Applied Gene Technology, Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Central 6, 1-1-1 Higashi, Tsukuba, Ibaraki 305-8566 (Japan); Kobayashi, Toshikatsu [Innovation Center for Startups, National Institute of Advanced Industrial Science and Technology, 2-2-2 Marunouchi, Chiyoda-ku, Tokyo 100-0005 (Japan); Janusys Corporation, 508, Saitama Industrial Technology Center, Skip City, 3-12-18 Kami-Aoki, Kawaguchi, Saitama 333-0844 (Japan); Sakai, Takafumi [Graduate School of Science and Engineering, Saitama University, 255 Shimo-Okubo, Sakura-ku, Saitama 338-8570 (Japan)

    2012-04-27

    Highlights: Black-Right-Pointing-Pointer Disulfide-rich peptide aptamer inhibits IL-6-dependent cell proliferation. Black-Right-Pointing-Pointer Disulfide bond of peptide aptamer is essential for its affinity to IL-6R. Black-Right-Pointing-Pointer Inhibitory effect of peptide depends on number and pattern of its disulfide bonds. -- Abstract: Several engineered protein scaffolds have been developed recently to circumvent particular disadvantages of antibodies such as their large size and complex composition, low stability, and high production costs. We previously identified peptide aptamers containing one or two disulfide-bonds as an alternative ligand to the interleukin-6 receptor (IL-6R). Peptide aptamers (32 amino acids in length) were screened from a random peptide library by in vitro peptide selection using the evolutionary molecular engineering method 'cDNA display'. In this report, the antagonistic activity of the peptide aptamers were examined by an in vitro competition enzyme-linked immunosorbent assay (ELISA) and an IL-6-dependent cell proliferation assay. The results revealed that a disulfide-rich peptide aptamer inhibited IL-6-dependent cell proliferation with similar efficacy to an anti-IL-6R monoclonal antibody.

  15. A disulfide bond in the TIM23 complex is crucial for voltage gating and mitochondrial protein import.

    Science.gov (United States)

    Ramesh, Ajay; Peleh, Valentina; Martinez-Caballero, Sonia; Wollweber, Florian; Sommer, Frederik; van der Laan, Martin; Schroda, Michael; Alexander, R Todd; Campo, María Luisa; Herrmann, Johannes M

    2016-08-15

    Tim17 is a central, membrane-embedded subunit of the mitochondrial protein import machinery. In this study, we show that Tim17 contains a pair of highly conserved cysteine residues that form a structural disulfide bond exposed to the intermembrane space (IMS). This disulfide bond is critical for efficient protein translocation through the TIM23 complex and for dynamic gating of its preprotein-conducting channel. The disulfide bond in Tim17 is formed during insertion of the protein into the inner membrane. Whereas the import of Tim17 depends on the binding to the IMS protein Mia40, the oxidoreductase activity of Mia40 is surprisingly dispensable for Tim17 oxidation. Our observations suggest that Tim17 can be directly oxidized by the sulfhydryl oxidase Erv1. Thus, import and oxidation of Tim17 are mediated by the mitochondrial disulfide relay, though the mechanism by which the disulfide bond in Tim17 is formed differs considerably from that of soluble IMS proteins. PMID:27502485

  16. An algorithmic approach to automated high-throughput identification of disulfide connectivity in proteins using tandem mass spectrometry.

    Science.gov (United States)

    Lee, Timothy; Singh, Rahul; Yen, Ten-Yang; Macher, Bruce

    2007-01-01

    Knowledge of the pattern of disulfide linkages in a protein leads to a better understanding of its tertiary structure and biological function. At the state-of-the-art, liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) can produce spectra of the peptides in a protein that are putatively joined by a disulfide bond. In this setting, efficient algorithms are required for matching the theoretical mass spaces of all possible bonded peptide fragments to the experimentally derived spectra to determine the number and location of the disulfide bonds. The algorithmic solution must also account for issues associated with interpreting experimental data from mass spectrometry, such as noise, isotopic variation, neutral loss, and charge state uncertainty. In this paper, we propose a algorithmic approach to high-throughput disulfide bond identification using data from mass spectrometry, that addresses all the aforementioned issues in a unified framework. The complexity of the proposed solution is of the order of the input spectra. The efficacy and efficiency of the method was validated using experimental data derived from proteins with with diverse disulfide linkage patterns.

  17. The road to the first, fully active and more stable human insulin variant with an additional disulfide bond.

    Science.gov (United States)

    Vinther, Tine N; Kjeldsen, Thomas B; Jensen, Knud J; Hubálek, František

    2015-11-01

    Insulin, a small peptide hormone, is crucial in maintaining blood glucose homeostasis. The stability and activity of the protein is directed by an intricate system involving disulfide bonds to stabilize the active monomeric species and by their non-covalent oligomerization. All known insulin variants in vertebrates consist of two peptide chains and have six cysteine residues, which form three disulfide bonds, two of them link the two chains and a third is an intra-chain bond in the A-chain. This classical insulin fold appears to have been conserved over half a billion years of evolution. We addressed the question whether a human insulin variant with four disulfide bonds could exist and be fully functional. In this review, we give an overview of the road to engineering four-disulfide bonded insulin analogs. During our journey, we discovered several active four disulfide bonded insulin analogs with markedly improved stability and gained insights into the instability of analogs with seven cysteine residues, importance of dimerization for stability, insulin fibril formation process, and the conformation of insulin binding to its receptor. Our results also open the way for new strategies in the development of insulin biopharmaceuticals. PMID:26382042

  18. Chemically exfoliated large-area two-dimensional flakes of molybdenum disulfide for device applications

    Directory of Open Access Journals (Sweden)

    Vivek Pachauri

    2013-09-01

    Full Text Available A solution-based exfoliation method for obtaining large-area two-dimensional flakes of molybdenum disulfide, followed by the fabrication of electrical devices is presented in this manuscript. The exfoliation method is based on the use of an aprotic solvent, namely, acetonitrile under mild sonication steps. In order to fabricate devices, a dielectrophoresis technique is used for transferring MoS2 flakes site-specifically on to the electrode pairs pre-written on the glass chips. The devices fabricated thus can be operated as chemical sensor in liquids while investigations under photo illumination indicate that such devices can also efficiently function as photodetectors.

  19. Efficient assembly of recombinant major histocompatibility complex class I molecules with preformed disulfide bonds

    DEFF Research Database (Denmark)

    Ostergaard Pedersen, L; Nissen, Mogens Holst; Hansen, N J;

    2001-01-01

    The expression of major histocompatibility class I (MHC-I) crucially depends upon the binding of appropriate peptides. MHC-I from natural sources are therefore always preoccupied with peptides complicating their purification and analysis. Here, we present an efficient solution to this problem...... suggests that de novo folding of denatured MHC-I molecules proceed efficiently if directed by preformed disulfide bond(s). Importantly, these molecules express serological epitopes and stain specific T cells; and they bind peptides specifically. Several denatured MHC-I heavy chains were analyzed and shown...

  20. Few-layer molybdenum disulfide transistors and circuits for high-speed flexible electronics

    OpenAIRE

    Cheng, Rui; Jiang, Shan; Chen, Yu; Liu, Yuan; Weiss, Nathan; Cheng, Hung-Chieh; Wu, Hao; Huang, Yu; Duan, Xiangfeng

    2014-01-01

    Two-dimensional layered materials, such as molybdenum disulfide, are emerging as an exciting material system for future electronics due to their unique electronic properties and atomically thin geometry. Here we report a systematic investigation of MoS2 transistors with optimized contact and device geometry, to achieve self-aligned devices with performance including an intrinsic gain over 30, an intrinsic cut-off frequency fT up to 42 GHz and a maximum oscillation frequency fMAX up to 50 GHz,...

  1. Dibromido(di-2-pyridyl disulfide-κ2N,N′zinc(II

    Directory of Open Access Journals (Sweden)

    Mario Wriedt

    2008-01-01

    Full Text Available The molecular structure of the title compound, [ZnBr2(C10H8N2S2], contains a seven-membered chelate ring in which the zinc atom is coordinated by two bromide ions and by the two pyridyl N atoms of a single 2,2′-dipyridyldisulfide (dpds ligand within a slightly distorted tetrahedron. As is usual for this type of complex, the disulfide group does not participate in zinc coordination. The chelate complexes are connected via weak intermolecular C—H...Br hydrogen bonding into chains, which extend in the [010] direction.

  2. The compromise of dynamic disulfide/thiol homeostasis as a biomarker of oxidative stress in trichloroethylene exposure.

    Science.gov (United States)

    Bal, C; Büyükşekerci, M; Koca, C; Ağış, E R; Erdoğan, S; Baran, P; Gündüzöz, M; Yilmaz, Öh

    2016-09-01

    In this study, we aimed to investigate disulfide/thiol homeostasis in trichloroethylene (TCE) exposure. The study was carried out in 30 nonsmoker TCE-exposed workers with a variety of occupations. Additionally, 30 healthy nonsmoker volunteers were recruited as the control group. TCE exposure was determined by measuring urinary trichloroacetic acid (TCA) concentration. Median urinary TCA levels of exposed workers (20.5 mg/L) were significantly higher than control subjects (5 mg/L). Thiol and disulfide concentrations were determined using a novel automated method. Disulfide/thiol ratio was significantly higher in the exposed group (p TCE-exposed workers. We predict that in TCE-exposed workers this disturbance can be a therapeutic target, and the efficiency of the treatment can easily be monitored by the novel method we used. PMID:26429930

  3. Characterization of intramolecular disulfide bonds and secondary modifications of the glycoprotein from viral hemorrhagic septicemia virus, a fish rhabdovirus

    DEFF Research Database (Denmark)

    Einer-Jensen, Katja; Nielsen, Thomas Krogh; Roepstorff, Peter;

    1998-01-01

    were analyzed by mass spectrometry before and after chemical reduction, and six disulfide bonds were identified: Cys29-Cys339, Cys44-Cys295, Cys90-Cys132, Cys172-Cys177, Cys195-Cys265, and Cys231-CyS236. Mass spectrometric analysis in combination with glycosidases allowed characterization of the glycan...... of the protein, The present study was initiated to identify the disulfide bonds and other structural aspects relevant to vaccine design. The N-terminal amino acid residue was identified as being a pyroglutamic acid, corresponding to Gln21 of the primary transcript, Peptides from endoproteinase-degraded G protein...... cysteine residues are situated at conserved positions, This finding suggests that there might be some common disulfide bonding pattern among the six rhabdoviruses....

  4. Influence of interface structures on the properties of molybdenum disulfide/graphene composites: A density functional theory study

    Energy Technology Data Exchange (ETDEWEB)

    Zan, Wenyan; Geng, Wei [State Key Laboratory of Applied Organic Chemistry, Department of Chemistry, Lanzhou University, Lanzhou 730000 (China); Liu, Huanxiang [School of Pharmacy, Lanzhou University, Lanzhou 730000 (China); Yao, Xiaojun, E-mail: xjyao@lzu.edu.cn [State Key Laboratory of Applied Organic Chemistry, Department of Chemistry, Lanzhou University, Lanzhou 730000 (China)

    2015-11-15

    Density functional theory calculations were performed to study the photocatalytic properties of molybdenum disulfide/graphene composites by analyzing the structure, electronic properties and optical properties of molybdenum disulfide/graphene composites. Three typical structures of molybdenum disulfide considered in our work include pristine molybdenum disulfide and molybdenum disulfide with mononiobium doping. They were then composited with graphene, N-doped graphene and graphene with epoxy, respectively. The characteristics of these composites (MoS{sub 2}/graphene, MoS{sub 2}/N-G, MoS{sub 2}/O-G and Nb–MoS{sub 2}/N-G) including binding energies, charge transfer, projected density of states, electron density and optical properties were calculated and analyzed. The binding energies of between MoS{sub 2} and graphene were related to the extent of charge transfer. The data of projected density of states, band structures and optical properties gave an explanation of the mechanism for significant photocatalytic activity of MoS{sub 2}/N-doped graphene and Nb-doped MoS{sub 2}/N-doped graphene composites. - Highlights: • MoS{sub 2}/graphene, MoS{sub 2}/N-doped graphene, MoS{sub 2}/graphene with epoxy and Nb-doped MoS{sub 2}/N-doped graphene were studied. • The electronic and optical properties of molybdenum disulfide/graphene composites were calculated. • MoS{sub 2}/N-doped graphene and Nb-doped MoS{sub 2}/N-doped graphene composites had better photocatalytic performance.

  5. Intracellular reduction/activation of a disulfide switch in thiosemicarbazone iron chelators.

    Science.gov (United States)

    Akam, Eman A; Chang, Tsuhen M; Astashkin, Andrei V; Tomat, Elisa

    2014-10-01

    Iron scavengers (chelators) offer therapeutic opportunities in anticancer drug design by targeting the increased demand for iron in cancer cells as compared to normal cells. Prochelation approaches are expected to avoid systemic iron depletion as chelators are liberated under specific intracellular conditions. In the strategy described herein, a disulfide linkage is employed as a redox-directed switch within the binding unit of an antiproliferative thiosemicarbazone prochelator, which is activated for iron coordination following reduction to the thiolate chelator. In glutathione redox buffer, this reduction event occurs at physiological concentrations and half-cell potentials. Consistent with concurrent reduction and activation, higher intracellular thiol concentrations increase cell susceptibility to prochelator toxicity in cultured cancer cells. The reduction of the disulfide switch and intracellular iron chelation are confirmed in cell-based assays using calcein as a fluorescent probe for paramagnetic ions. The resulting low-spin Fe(III) complex is identified in intact Jurkat cells by EPR spectroscopy measurements, which also document a decreased concentration of active ribonucleotide reductase following exposure to the prochelator. Cell viability and fluorescence-based assays show that the iron complex presents low cytotoxicity and does not participate in intracellular redox chemistry, indicating that this antiproliferative chelation strategy does not rely on the generation of reactive oxygen species. PMID:25100578

  6. N,N'-Dithiobisphthalimide, a disulfide aromatic compound, is a potent spermicide agent in humans.

    Science.gov (United States)

    Florez, Martha; Díaz, Emilce S; Brito, Iván; González, Jorge; Morales, Patricio

    2011-12-01

    Several studies have shown that users of vaginal preparations containing nonoxynol-9 (N-9) are at a high risk for sexually transmitted diseases, including HIV. Therefore, there is a great interest in identifying compounds that can specifically inhibit sperm without damaging the vaginal lining, possess a powerful spermicide activity, and can be used in contraceptive vaginal preparations to replace N-9. In this work, we studied the spermostatic and/or spermicidal activity of five non-detergent, disulfide compounds on human sperm, HeLa cells, and Lactobacillus acidophilus. The motility and viability of human sperm in semen and culture medium was evaluated after treatment with different concentrations of the disulfide compounds (2.5 - 100 µM). In addition, we evaluated the cytotoxic effect on HeLa cells and L. acidophilus. We identified compound 101, N,N'-dithiobisphthalimide (No. CAS 7764-30-9), as the most effective molecule. It has a half maximal effective concentration (EC(50)) of 8 µM and a minimum effective concentration (defined as the concentration that immobilizes 100 percent of the sperm in 20 sec) of 24 µM. At these concentrations, compound 101 does not affect the viability of the sperm, HeLa cells, or L. acidophilus. Our results indicate that dithiobisphthalimide has a potent spermostatic, irreversible effect with no toxic effects on HeLa cells and L. acidophilus. PMID:21942567

  7. Intracellular reduction/activation of a disulfide switch in thiosemicarbazone iron chelators

    Science.gov (United States)

    Akam, Eman A.; Chang, Tsuhen M.; Astashkin, Andrei V.

    2014-01-01

    Iron scavengers (chelators) offer therapeutic opportunities in anticancer drug design by targeting the increased demand for iron in cancer cells as compared to normal cells. Prochelation approaches are expected to avoid systemic iron depletion as chelators are liberated under specific intracellular conditions. In the strategy described herein, a disulfide linkage is employed as a redox-directed switch within the binding unit of an antiproliferative thiosemicarbazone prochelator, which is activated for iron coordination following reduction to the thiolate chelator. In glutathione redox buffer, this reduction event occurs at physiological concentrations and half-cell potentials. Consistent with concurrent reduction and activation, higher intracellular thiol concentrations increase cell susceptibility to prochelator toxicity in cultured cancer cells. The reduction of the disulfide switch and intracellular iron chelation are confirmed in cell-based assays using calcein as a fluorescent probe for paramagnetic ions. The resulting low-spin Fe(III) complex is identified in intact Jurkat cells by EPR spectroscopy measurements, which also document a decreased concentration of active ribonucleotide reductase following exposure to the prochelator. Cell viability and fluorescence-based assays show that the iron complex presents low cytotoxicity and does not participate in intracellular redox chemistry, indicating that this antiproliferative chelation strategy does not rely on the generation of reactive oxygen species. PMID:25100578

  8. Identification of intra- and intermolecular disulfide bridges in the multidrug resistance transporter ABCG2

    DEFF Research Database (Denmark)

    Henriksen, Ulla Birk; Fog, Jacob U; Litman, Thomas;

    2005-01-01

    cysteines predicted to be on the extracellular face of ABCG2. Upon mutation of Cys-592 or Cys-608 to alanine (C592A and C608A), ABCG2 migrated as a dimer in SDS-PAGE under non-reducing conditions; however, mutation of Cys-603 to Ala (C603A) caused the transporter to migrate as a single monomeric band......ABCG2 is an ATP binding cassette (ABC) half-transporter that plays a key role in multidrug resistance to chemotherapy. ABCG2 is believed to be a functional homodimer that has been proposed to be linked by disulfide bridges. We have investigated the structural and functional role of the only three....... Despite this change, C603A displayed efficient membrane targeting and preserved transport function. Because the transporter migrated as a dimer in SDS-PAGE, when only Cys-603 was present (C592A-C608A), the data suggest that Cys-603 forms a symmetrical intermolecular disulfide bridge in the ABCG2 homodimer...

  9. Oxidative folding and reductive activities of EhPDI, a protein disulfide isomerase from Entamoeba histolytica.

    Science.gov (United States)

    Mares, Rosa E; Magaña, Paloma D; Meléndez-López, Samuel G; Licea, Alexei F; Cornejo-Bravo, José M; Ramos, Marco A

    2009-09-01

    PDI enzymes are oxidoreductases that catalyze oxidation, reduction and isomerization of disulfide bonds in polypeptide substrates. We have previously identified an E. histolytica PDI enzyme (EhPDI) that exhibits oxidase activity in vivo. However, little is known about the specific role of its redox-related structural features on the enzymatic activity. Here, we have studied the in vivo oxidative folding of EhPDI by mutagenic analysis and functional complementation assays as well as the in vitro oxidative folding and reductive activities by comparative kinetics using functional homologues in standard assays. We have found that the active-site cysteine residues of the functional domains (Trx-domains) are essential for catalysis of disulfide bond formation in polypeptides and proteins, such as the bacterial alkaline phosphatase. Furthermore, we have shown that the recombinant EhPDI enzyme has some typical properties of PDI enzymes: oxidase and reductase activities. These activities were comparable to those observed for other functional equivalents, such as bovine PDI or bacterial thioredoxin, under the same experimental conditions. These findings will be helpful for further studies intended to understand the physiological role of EhPDI.

  10. Maternal nicotine exposure leads to impaired disulfide bond formation and augmented endoplasmic reticulum stress in the rat placenta.

    Directory of Open Access Journals (Sweden)

    Michael K Wong

    Full Text Available Maternal nicotine exposure has been associated with many adverse fetal and placental outcomes. Although underlying mechanisms remain elusive, recent studies have identified that augmented endoplasmic reticulum (ER stress is linked to placental insufficiency. Moreover, ER function depends on proper disulfide bond formation--a partially oxygen-dependent process mediated by protein disulfide isomerase (PDI and ER oxidoreductases. Given that nicotine compromised placental development in the rat, and placental insufficiency has been associated with poor disulfide bond formation and ER stress, we hypothesized that maternal nicotine exposure leads to both placental ER stress and impaired disulfide bond formation. To test this hypothesis, female Wistar rats received daily subcutaneous injections of either saline (vehicle or nicotine bitartrate (1 mg/kg for 14 days prior to mating and during pregnancy. Placentas were harvested on embryonic day 15 for analysis. Protein and mRNA expression of markers involved in ER stress (e.g., phosphorylated eIF2α, Grp78, Atf4, and CHOP, disulfide bond formation (e.g., PDI, QSOX1, VKORC1, hypoxia (Hif1α, and amino acid deprivation (GCN2 were quantified via Western blot and/or Real-time PCR. Maternal nicotine exposure led to increased expression of Grp78, phosphorylated eIF2α, Atf4, and CHOP (p<0.05 in the rat placenta, demonstrating the presence of augmented ER stress. Decreased expression of PDI and QSOX1 (p<0.05 reveal an impaired disulfide bond formation pathway, which may underlie nicotine-induced ER stress. Finally, elevated expression of Hif1α and GCN2 (p<0.05 indicate hypoxia and amino acid deprivation in nicotine-exposed placentas, respectively, which may also cause impaired disulfide bond formation and augmented ER stress. This study is the first to link maternal nicotine exposure with both placental ER stress and disulfide bond impairment in vivo, providing novel insight into the mechanisms underlying

  11. Co-factor insertion and disulfide bond requirements for twin-arginine translocase-dependent export of the Bacillus subtilis Rieske protein QcrA

    NARCIS (Netherlands)

    Goosens, Vivianne J; Monteferrante, Carmine G; van Dijl, Jan Maarten

    2014-01-01

    Background: The Rieske protein QcrA was recently shown to be exported by twin-arginine translocation (Tat) in Bacillus subtilis. Results: QcrA has disulfide bond and co-factor requirements for effective Tat-dependent translocation. Conclusion: A hierarchy exists between disulfide bonding and co-fact

  12. Identification, activity and disulfide connectivity of C-di-GMP regulating proteins in Mycobacterium tuberculosis.

    Directory of Open Access Journals (Sweden)

    Kajal Gupta

    Full Text Available C-di-GMP, a bacterial second messenger plays a key role in survival and adaptation of bacteria under different environmental conditions. The level of c-di-GMP is regulated by two opposing activities, namely diguanylate cyclase (DGC and phosphodiesterase (PDE-A exhibited by GGDEF and EAL domain, respectively in the same protein. Previously, we reported a bifunctional GGDEF-EAL domain protein, MSDGC-1 from Mycobacterium smegmatis showing both these activities (Kumar and Chatterji, 2008. In this current report, we have identified and characterized the homologous protein from Mycobacterium tuberculosis (Rv 1354c named as MtbDGC. MtbDGC is also a bifunctional protein, which can synthesize and degrade c-di-GMP in vitro. Further we expressed Mtbdgc in M. smegmatis and it was able to complement the MSDGC-1 knock out strain by restoring the long term survival of M. smegmatis. Another protein Rv 1357c, named as MtbPDE, is an EAL domain protein and degrades c-di-GMP to pGpG in vitro. Rv1354c and 1357c have seven cysteine amino acids in their sequence, distributed along the full length of the protein. Disulfide bonds play an important role in stabilizing protein structure and regulating protein function. By proteolytic digestion and mass spectrometric analysis of MtbDGC, connectivity between cysteine pairs Cys94-Cys584, Cys2-Cys479 and Cys429-Cys614 was determined, whereas the third cysteine (Cys406 from N terminal was found to be free in MtbDGC protein, which was further confirmed by alkylation with iodoacetamide labeling. Bioinformatics modeling investigations also supported the pattern of disulfide connectivity obtained by Mass spectrometric analysis. Cys406 was mutated to serine by site directed mutagenesis and the mutant MtbC406S was not found to be active and was not able to synthesize or degrade c-di-GMP. The disulfide connectivity established here would help further in understanding the structure - function relationship in MtbDGC.

  13. Post-translational disulfide modifications in cell signaling--role of inter-protein, intra-protein, S-glutathionyl, and S-cysteaminyl disulfide modifications in signal transmission.

    Science.gov (United States)

    O'Brian, Catherine A; Chu, Feng

    2005-05-01

    Cell signaling entails a host of post-translational modifications of effector-proteins. These modifications control signal transmission by regulating the activity, localization or half-life of the effector-protein. Prominent oxidative modifications induced by cell-signaling reactive oxygen species (ROS) are cysteinyl modifications such as S-nitrosylation, sulfenic acid and disulfide formation. Disulfides protect protein sulfhydryls against oxidative destruction and simultaneously influence cell signaling by engaging redox-regulatory sulfhydryls in effector-proteins. The types of disulfides implicated in signaling span (1) protein S-glutathionylation, e.g. as a novel mode of Ras activation through S-glutathionylation at Cys-118 in response to a hydrogen-peroxide burst, (2) intra-protein disulfides, e.g. in the regulation of the stability of the protein phosphatase Cdc25C by hydrogen-peroxide, (3) inter-protein disulfides, e.g. in the hydrogen peroxide-mediated inactivation of receptor protein-tyrosine phosphatase alpha (RPTPalpha) by dimerization and (4) protein S-cysteaminylation by cystamine. Cystamine is a byproduct of pantetheinase-catalyzed pantothenic acid recycling from pantetheine for biosynthesis of Coenzyme A (CoA), a ubiquitous and metabolically indispensable cofactor. Cystamine inactivates protein kinase C-epsilon (PKCepsilon), gamma-glutamylcysteine synthetase and tissue transglutaminase by S-cysteaminylation-triggered mechanisms. The importance of protein S-cysteaminylation in signal transmission in vivo is evident from the ability of cystamine administration to rescue the intestinal inflammatory-response deficit of pantetheinase knockout mice. These mice lack the predominant epithelial pantetheinase isoform and have sharply reduced levels of cystamine/cysteamine in epithelial tissues. In addition, intraperitoneal administration of cystamine significantly delays neurodegenerative pathogenesis in a Huntington's disease mouse model. Thus, cystamine may

  14. Identification of Thioredoxin Disulfide Targets Using a Quantitative Proteomics Approach Based on Isotope-Coded Affinity Tags

    DEFF Research Database (Denmark)

    Hägglund, Per; Bunkenborg, Jakob; Maeda, Kenji;

    2008-01-01

    , protein extract of embryos from germinated barley seeds was treated +/- Trx, and thiols released from target protein disulfides were irreversibly blocked with iodoacetamide. The remaining cysteine residues in the Trx-treated and the control (-Trx) samples were then chemically reduced and labeled...

  15. Disulfide-Based Diblock Copolymer Worm Gels: A Wholly-Synthetic Thermoreversible 3D Matrix for Sheet-Based Cultures.

    Science.gov (United States)

    Simon, Karen A; Warren, Nicholas J; Mosadegh, Bobak; Mohammady, Marym R; Whitesides, George M; Armes, Steven P

    2015-12-14

    It is well-known that 3D in vitro cell cultures provide a much better model than 2D cell cultures for understanding the in vivo microenvironment of cells. However, significant technical challenges in handling and analyzing 3D cell cultures remain, which currently limits their widespread application. Herein, we demonstrate the application of wholly synthetic thermoresponsive block copolymer worms in sheet-based 3D cell culture. These worms form a soft, free-standing gel reversibly at 20-37 °C, which can be rapidly converted into a free-flowing dispersion of spheres on cooling to 5 °C. Functionalization of the worms with disulfide groups was found to be essential for ensuring sufficient mechanical stability of these hydrogels to enable long-term cell culture. These disulfide groups are conveniently introduced via statistical copolymerization of a disulfide-based dimethacrylate under conditions that favor intramolecular cyclization and subsequent thiol/disulfide exchange leads to the formation of reversible covalent bonds between adjacent worms within the gel. This new approach enables cells to be embedded within micrometer-thick slabs of gel with good viability, permits cell culture for at least 12 days, and facilitates recovery of viable cells from the gel simply by incubating the culture in buffer at 4 °C (thus, avoiding the enzymatic degradation required for cell harvesting when using commercial protein-based gels, such as Matrigel). PMID:26509930

  16. Formation of disulfide bridges by a single-chain Fv antibody in the reducing ectopic environment of the plant cytosol

    NARCIS (Netherlands)

    Schouten, A.; Roosien, J.; Bakker, J.; Schots, A.

    2002-01-01

    Disulfide bridge formation in the reducing environment of the cytosol is considered a rare event and is mostly linked to inactivation of protein activity. In this report the in vivo redox state of a single-chain Fv (scFv) antibody fragment in the plant cytosol was investigated. The scFv antibody fra

  17. A dielectric barrier discharge terminally inactivates RNase A by oxidizing sulfur-containing amino acids and breaking structural disulfide bonds

    Science.gov (United States)

    Lackmann, J.-W.; Baldus, S.; Steinborn, E.; Edengeiser, E.; Kogelheide, F.; Langklotz, S.; Schneider, S.; Leichert, L. I. O.; Benedikt, J.; Awakowicz, P.; Bandow, J. E.

    2015-12-01

    RNases are among the most stable proteins in nature. They even refold spontaneously after heat inactivation, regaining full activity. Due to their stability and universal presence, they often pose a problem when experimenting with RNA. We investigated the capabilities of nonthermal atmospheric-pressure plasmas to inactivate RNase A and studied the inactivation mechanism on a molecular level. While prolonged heating above 90 °C is required for heat inactivating RNase A, direct plasma treatment with a dielectric barrier discharge (DBD) source caused permanent inactivation within minutes. Circular dichroism spectroscopy showed that DBD-treated RNase A unfolds rapidly. Raman spectroscopy indicated methionine modifications and formation of sulfonic acid. A mass spectrometry-based analysis of the protein modifications that occur during plasma treatment over time revealed that methionine sulfoxide formation coincides with protein inactivation. Chemical reduction of methionine sulfoxides partially restored RNase A activity confirming that sulfoxidation is causal and sufficient for RNase A inactivation. Continued plasma exposure led to over-oxidation of structural disulfide bonds. Using antibodies, disulfide bond over-oxidation was shown to be a general protein inactivation mechanism of the DBD. The antibody’s heavy and light chains linked by disulfide bonds dissociated after plasma exposure. Based on their ability to inactivate proteins by oxidation of sulfur-containing amino acids and over-oxidation of disulfide bonds, DBD devices present a viable option for inactivating undesired or hazardous proteins on heat or solvent-sensitive surfaces.

  18. Disulfide assignment of the C-terminal cysteine knot of agouti-related protein (AGRP) by direct sequencing analysis.

    Science.gov (United States)

    Young, Y; Zeni, L; Rosenfeld, R D; Stark, K L; Rohde, M F; Haniu, M

    1999-12-01

    We have assigned the disulfide structure of Md-65 agouti-related protein (Md65-AGRP) using differential reduction and alkylation followed by direct sequencing analysis. The mature human AGRP is a single polypeptide chain of 112 amino acid residues, consisting of an N-terminal acidic region and a unique C-terminal cysteine-rich domain. The C-terminal domain, a 48 amino acid peptide named Md65-AGRP, was expressed in Escherichia coil cells and refolded under different conditions from the mature recombinant protein. The disulfide bonds in the cystine knot structure of Md65-AGRP were partially reduced using tris(2-carboxyethyl) phosphine (TCEP) under acidic conditions, followed by alkylation with N-ethylmaleimide (NEM). The procedure generated several isoforms with varying degrees of NEM alkylation. The multiple forms of Md65-AGRP generated by partial reduction and NEM modification were then completely reduced and carboxymethylated to identify unreactive disulfide bonds. Differentially labeled Md65-AGRP were directly sequenced and analyzed by MALDI mass spectrometry. The results confirmed that Md65-AGRP contained the same disulfide structure as that of Md5-AGRP reported previously [Bures, E. J., Hui, J. O., Young, Y. et al. (1998) Biochemistry 37, 12172-12177].

  19. Activity assays of mammalian thioredoxin and thioredoxin reductase: fluorescent disulfide substrates, mechanisms, and use with tissue samples.

    Science.gov (United States)

    Montano, Sergio J; Lu, Jun; Gustafsson, Tomas N; Holmgren, Arne

    2014-03-15

    Thioredoxin (Trx) is a protein disulfide reductase that, together with nicotinamide adenine dinucleotide phosphate (NADPH) and thioredoxin reductase (TrxR), controls oxidative stress or redox signaling via thiol redox control. Human cytosolic Trx1 has Cys32 and Cys35 as the active site and three additional cysteine residues (Cys62, Cys69, and Cys73), which by oxidation generates inactive Cys62 to Cys69 two-disulfide Trx. This, combined with TrxR with a broad substrate specificity, complicates assays of mammalian Trx and TrxR. We sought to understand the autoregulation of Trx and TrxR and to generate new methods for quantification of Trx and TrxR. We optimized the synthesis of two fluorescent substrates, di-eosin-glutathione disulfide (Di-E-GSSG) and fluorescein isothiocyanate-labeled insulin (FiTC-insulin), which displayed higher fluorescence on disulfide reduction. Di-E-GSSG showed a very large increase in fluorescence quantum yield but had a relatively low affinity for Trx and was also a weak direct substrate for TrxR, in contrast to GSSG. FiTC-insulin was used to develop highly sensitive assays for TrxR and Trx. Reproducible conditions were developed for reactivation of modified Trx, commonly present in frozen or oxidized samples. Trx in cell extracts and tissue samples, including plasma and serum, were subsequently analyzed, showing highly reproducible results and allowing measurement of trace amounts of Trx.

  20. Human Islet Amyloid Polypeptide N-Terminus Fragment Self-Assembly: Effect of Conserved Disulfide Bond on Aggregation Propensity

    Science.gov (United States)

    Ilitchev, Alexandre I.; Giammona, Maxwell J.; Do, Thanh D.; Wong, Amy G.; Buratto, Steven K.; Shea, Joan-Emma; Raleigh, Daniel P.; Bowers, Michael T.

    2016-06-01

    Amyloid formation by human islet amyloid polypeptide (hIAPP) has long been implicated in the pathogeny of type 2 diabetes mellitus (T2DM) and failure of islet transplants, but the mechanism of IAPP self-assembly is still unclear. Numerous fragments of hIAPP are capable of self-association into oligomeric aggregates, both amyloid and non-amyloid in structure. The N-terminal region of IAPP contains a conserved disulfide bond between cysteines at position 2 and 7, which is important to hIAPP's in vivo function and may play a role in in vitro aggregation. The importance of the disulfide bond in this region was probed using a combination of ion mobility-based mass spectrometry experiments, molecular dynamics simulations, and high-resolution atomic force microscopy imaging on the wildtype 1-8 hIAPP fragment, a reduced fragment with no disulfide bond, and a fragment with both cysteines at positions 2 and 7 mutated to serine. The results indicate the wildtype fragment aggregates by a different pathway than either comparison peptide and that the intact disulfide bond may be protective against aggregation due to a reduction of inter-peptide hydrogen bonding.

  1. Fast and efficient green synthesis of thiosulfonate S-esters by microwave-supported permanganate oxidation of symmetrical disulfides

    DEFF Research Database (Denmark)

    Thi, Luu Thi Xuan; Thi Nguyen, Thao-Tran; Le, Thach Ngoc;

    2015-01-01

    Potassium permanganate absorbed on copper(II) sulfate pentahydrate has been found to be an efficient, inexpensive, and green oxidation agent for the synthesis of “symmetrical” thiosulfonate S-esters by oxidation of the corresponding symmetrical disulfides. The oxidation reactions were carried out...

  2. Solubility of disulfide-bonded proteins in the cytoplasm of Escherichia coli and its "oxidizing" mutant

    Institute of Scientific and Technical Information of China (English)

    Sheng Xiong; Yi-Fei Wang; Xiang-Rong Ren; Bing Li; Mei-Ying Zhang; Yong Luo; Ling Zhang; Qiu-Ling Xie; Kuan-Yuan Su

    2005-01-01

    AIM: To study the influence of redox environment of Escherichia coli ( E. coli) cytoplasm on disulfide bond formation of recombinant proteins.METHODS: Bovine fibroblast growth factor (BbFGF) was selected as a model of simple proteins with a single disulfide bond and free cysteines. Anti-HBsAg single-chain Fv (HBscFv), an artificial multidomain protein, was selected as the model molecule of complex protein with 2 disulfide bonds. A BbFGF-producing plasmid, pJN-BbFGF,and a HBscFv producing-plasmid, pQE-HBscFv, were constructed and transformed into E. coli strains BL21(DE3)and M15[pREP4]respectively. At the same time, both plasmids were transformedinto a reductase-deficient host strain, E. coli Origami(DE3). The 4 recombinant E. coli strains were cultured and the target proteins were purified. Solubility and bioactivity of recombinant BbFGF and HBscFv produced in different host strains were analyzed and compared respectively.RESULTS: All recombinant E. colistrains could efficiently produce target proteins. The level of BbFGF in BL21(DE3)was 15-23% of the total protein, and was 5-10% in Origami (DE3). In addition, 65% of the BbFGF produced in BL21(DE3) formed into inclusion body in the cytoplasm,and all the target proteins became soluble in Origami (DE3). The bioactivity of BbFGF purified from Origami(DE3)was higher than its counterpart from BL21(DE3). The ED50of BbFGF from Origami(DE3) and BL21(DE3) was 1.6 μg/L and 2.2 μg/L, respectively. Both HBscFv formed into inclusion body in the cytoplasm of M15[pQE-HBscFv] or Origami[pQE-HBscFv]. But the supernatant of Origami[pQE-HBscFv] lysate displayed weak bioactivity and its counterpart from M15[pQE-HBscFv] did not display any bioactivity. The soluble HBscFv in Origami[pQE-HBscFv]was purified to be 1-2 mg/L and its affinity constant was determined to be 2.62×107 mol/L. The yield of native HBscFv refolded from indusion body in M15[pQE-H Fv] was30-35 mg/L and the affinity constant was 1.98×107 mol/L.There was no

  3. High-performance molybdenum disulfide field-effect transistors with spin tunnel contacts.

    Science.gov (United States)

    Dankert, André; Langouche, Lennart; Kamalakar, Mutta Venkata; Dash, Saroj Prasad

    2014-01-28

    Molybdenum disulfide has recently emerged as a promising two-dimensional semiconducting material for nanoelectronic, optoelectronic, and spintronic applications. Here, we investigate the field-effect transistor behavior of MoS2 with ferromagnetic contacts to explore its potential for spintronics. In such devices, we elucidate that the presence of a large Schottky barrier resistance at the MoS2/ferromagnet interface is a major obstacle for the electrical spin injection and detection. We circumvent this problem by a reduction in the Schottky barrier height with the introduction of a thin TiO2 tunnel barrier between the ferromagnet and MoS2. This results in an enhancement of the transistor on-state current by 2 orders of magnitude and an increment in the field-effect mobility by a factor of 6. Our magnetoresistance calculation reveals that such integration of ferromagnetic tunnel contacts opens up the possibilities for MoS2-based spintronic devices. PMID:24377305

  4. Disulfide bond reduction-triggered molecular hydrogels of folic acid-Taxol conjugates.

    Science.gov (United States)

    Yang, Chengbiao; Li, Dongxia; Fengzhao, Qianqi; Wang, Lianyong; Wang, Ling; Yang, Zhimou

    2013-09-25

    Molecular hydrogels of therapeutic agents are a novel kind of self-delivery system that can sustain release of drugs or pro-drugs. We have previously developed a molecular hydrogelator of folic acid (FA)-Taxol conjugate triggered by phosphatase. In this paper, we report a novel molecular hydrogelator system of FA-Taxol conjugates with improved synthetic strategy. The hydrogels are formed by the reduction of disulfide bond by glutathione (GSH). These hydrogels could sustain release of Taxol through ester bond hydrolysis. Compared with intravenous (i.v.) injection of clinically used Taxol® with four times the dosage, our hydrogel could inhibit tumor growth more efficiently by a single dose of intra-tumor (i.t.) administration. These observations suggested the big potential of this novel gelation system of Taxol for cancer therapy.

  5. Simultaneous Disulfide and Boronic Acid Ester Exchange in Dynamic Combinatorial Libraries

    Directory of Open Access Journals (Sweden)

    Sanna L. Diemer

    2015-09-01

    Full Text Available Dynamic combinatorial chemistry has emerged as a promising tool for the discovery of complex receptors in supramolecular chemistry. At the heart of dynamic combinatorial chemistry are the reversible reactions that enable the exchange of building blocks between library members in dynamic combinatorial libraries (DCLs ensuring thermodynamic control over the system. If more than one reversible reaction operates in a single dynamic combinatorial library, the complexity of the system increases dramatically, and so does its possible applications. One can imagine two reversible reactions that operate simultaneously or two reversible reactions that operate independently. Both these scenarios have advantages and disadvantages. In this contribution, we show how disulfide exchange and boronic ester transesterification can function simultaneous in dynamic combinatorial libraries under appropriate conditions. We describe the detailed studies necessary to establish suitable reaction conditions and highlight the analytical techniques appropriate to study this type of system.

  6. Controlling the work function of molybdenum disulfide by in situ metal deposition

    Science.gov (United States)

    Zhou, Peng; Song, Xiongfei; Yan, Xiao; Liu, Chunsen; Chen, Lin; Sun, Qingqing; Zhang, David Wei

    2016-08-01

    Control of the work function of molybdenum disulfide (MoS2) under ultrathin metal was investigated using in situ metal deposition and direct ultraviolet photoelectron spectroscopy measurement in an ultra-high vacuum system. When the metal thickness turned from two dimensional into bulk, the work function was also raised up at the nickel‑MoS2 interface, barely changed at the titanium‑MoS2 interface and lowered at the hafnium‑MoS2 interface. Meanwhile, the mechanisms of charge transfer and band alignment with metal deposition were also discussed. The Schottky barrier at metal‑MoS2 interfaces could be tailored by both types and thicknesses of deposited metal. The low work function metal was a good indicator for MoS2 contact electrodes. It paved the way towards future high performance MoS2 device applications.

  7. Dissecting molecular interactions involved in recognition of target disulfides by the barley thioredoxin system

    DEFF Research Database (Denmark)

    Björnberg, Olof; Maeda, Kenji; Svensson, Birte;

    2012-01-01

    -amylase/subtilisin inhibitor (BASI), two loops in barley thioredoxin h2 (HvTrxh2), containing an invariant cis-proline (86EAMP89) and a conserved glycine (104VGA106), surround the active site cysteines ( 45WCGPC49) and contribute to binding of BASI through backbone-backbone hydrogen bonds [Maeda, K., Hägglund, P., Finnie, C...... retained catalytic properties, with the exception of a 3-fold increased activity toward BASI. From the 104VGA106 loop, a backbone hydrogen bond donated by A106 appears to be important for target disulfide recognition as A106P lost 90% activity toward BASI but was efficiently recycled by thioredoxin...... reductase. The findings support important roles in target recognition of backbone-backbone hydrogen bond and electrostatic interactions and are discussed in relation to earlier structural and functional studies of thioredoxins and related proteins. © 2012 American Chemical Society....

  8. Molybdenum Disulfide as a Protection Layer and Catalyst for Gallium Indium Phosphide Solar Water Splitting Photocathodes

    Energy Technology Data Exchange (ETDEWEB)

    Britto, Reuben J.; Benck, Jesse D.; Young, James L.; Hahn, Christopher; Deutsch, Todd G.; Jaramillo, Thomas F.

    2016-06-02

    Gallium indium phosphide (GaInP2) is a semiconductor with promising optical and electronic properties for solar water splitting, but its surface stability is problematic as it undergoes significant chemical and electrochemical corrosion in aqueous electrolytes. Molybdenum disulfide (MoS2) nanomaterials are promising to both protect GaInP2 and to improve catalysis since MoS2 is resistant to corrosion and also possesses high activity for the hydrogen evolution reaction (HER). In this work, we demonstrate that GaInP2 photocathodes coated with thin MoS2 surface protecting layers exhibit excellent activity and stability for solar hydrogen production, with no loss in performance (photocurrent onset potential, fill factor, and light limited current density) after 60 hours of operation. This represents a five-hundred fold increase in stability compared to bare p-GaInP2 samples tested in identical conditions.

  9. Transient existence of crystalline lithium disulfide Li2S2 in a lithium-sulfur battery

    Science.gov (United States)

    Paolella, Andrea; Zhu, Wen; Marceau, Hugues; Kim, Chi-su; Feng, Zimin; Liu, Dongqiang; Gagnon, Catherine; Trottier, Julie; Abdelbast, Guerfi; Hovington, Pierre; Vijh, Ashok; Demopoulos, George P.; Armand, Michel; Zaghib, Karim

    2016-09-01

    Crystalline lithium disulfide (Li2S2) is identified, for the first time, as a transient species in the lithium-sulfur cell, by using an operando X-ray diffraction (XRD) technique. The observed XRD pattern precisely matches with the predicted pattern based on the density function theory. The formation of Li2S2 crystals is repetitively found in the highly concentrated (7 M Li+) electrolyte at high voltage region (>2 V) near the end of the first charge cycle and before the end of the second discharge cycle. These conditions indicate that crystalline Li2S2 exists in the non-equilibrium regime. The formation of crystalline Li2S2 under only the specified conditions suggests that it is not formed as an intermediate discharge product, contrary to what is generally believed, but as a transient species by the disproportionation reaction from higher order polysulfides which is facilitated by the "solvent-in-salt" conditions.

  10. Increase Renaturation Yield of Reteplase Using the Recombinant Human Protein Disulfide Isomerase

    Institute of Scientific and Technical Information of China (English)

    Zhao Youchun(赵友春); Wang Ge; Kong Yang; Wang Yanbing; Zhang Changkai; Chen Chao; Liang Bufeng

    2004-01-01

    Reteplase, the recombinant type of novel tissue plasminogen activator (t-PA) variant, is a promising thrombolytics in clinics. Expressed in the form of an inclusion body, reteplase consists of about 40 % of the total intracellular proteins of Escherichia coli. The recombinant human protein disulfide isomerase (rhPDI) is used to increase the chance for the correct matching of the 18 hydrosulfide groups of the reteplase molecule in the renaturation process and it increase is the reteplase renaturation yield from 1%~2% to 15%~20% with a the purity aboue 99% and the specific activity of 5(105 IU/mg is reached. This novel method can reduce significantly the cost of production.

  11. Origin of Structural Transformation in Mono- and Bi-Layered Molybdenum Disulfide

    Science.gov (United States)

    Sun, Xiaoli; Wang, Zhiguo; Li, Zhijie; Fu, Y. Q.

    2016-05-01

    Mono- and multi-layered molybdenum disulfide (MoS2) is considered to be one of the next generation anode materials for rechargeable ion batteries. Structural transformation from trigonal prismatic (2H) to octahedral (1T) upon lithium or sodium intercalation has been in-situ observed experimentally using transmission electron microscope during studies of their electrochemical dynamics processes. In this work, we explored the fundamental mechanisms of this structural transformation in both mono- and bi-layered MoS2 using density functional theory. For the intercalated MoS2, the Li and Na donate their electrons to the MoS2. Based on the theoretical analysis, we confirmed that, for the first time, electron transfer is dominant in initiating this structural transformation, and the results provide an in-depth understanding of the transformation mechanism induced by the electron doping. The critical values of electron concentrations for this structural transformation are decreased with increasing the layer thickness.

  12. Bile salt-induced intermolecular disulfide bond formation activates Vibrio cholerae virulence.

    Science.gov (United States)

    Yang, Menghua; Liu, Zhi; Hughes, Chambers; Stern, Andrew M; Wang, Hui; Zhong, Zengtao; Kan, Biao; Fenical, William; Zhu, Jun

    2013-02-01

    To be successful pathogens, bacteria must often restrict the expression of virulence genes to host environments. This requires a physical or chemical marker of the host environment as well as a cognate bacterial system for sensing the presence of a host to appropriately time the activation of virulence. However, there have been remarkably few such signal-sensor pairs identified, and the molecular mechanisms for host-sensing are virtually unknown. By directly applying a reporter strain of Vibrio cholerae, the causative agent of cholera, to a thin layer chromatography (TLC) plate containing mouse intestinal extracts, we found two host signals that activate virulence gene transcription. One of these was revealed to be the bile salt taurocholate. We then show that a set of bile salts cause dimerization of the transmembrane transcription factor TcpP by inducing intermolecular disulfide bonds between cysteine (C)-207 residues in its periplasmic domain. Various genetic and biochemical analyses led us to propose a model in which the other cysteine in the periplasmic domain, C218, forms an inhibitory intramolecular disulfide bond with C207 that must be isomerized to form the active C207-C207 intermolecular bond. We then found bile salt-dependent effects of these cysteine mutations on survival in vivo, correlating to our in vitro model. Our results are a demonstration of a mechanism for direct activation of the V. cholerae virulence cascade by a host signal molecule. They further provide a paradigm for recognition of the host environment in pathogenic bacteria through periplasmic cysteine oxidation.

  13. CYP-independent inhibition of platelet aggregation in rabbits by a mixed disulfide conjugate of clopidogrel.

    Science.gov (United States)

    Zhang, H; Lauver, D A; Hollenberg, P F

    2014-12-01

    Dual antiplatelet therapy with clopidogrel and aspirin has been the standard of care in the United States for patients with acute coronary syndromes (ACS) and/or undergoing percutaneous coronary interventions (PCI). However, the effectiveness of clopidogrel varies significantly among different sub-populations due to inter-individual variability. In this study we examined the antiplatelet potential of a novel mixed disulfide conjugate of clopidogrel with the aim to overcome the inter-individual variability. In the metabolic studies using human liver microsomes and cDNA-expressed P450s, we confirmed that multiple P450s are involved in the bioactivation of 2-oxoclopidogrel to H4, one of the diastereomers of the pharmacologically active metabolite (AM) possessing antiplatelet activity. Results from kinetic studies demonstrated that 2C19 is the most active in converting 2-oxoclopidogrel to H4 with a catalytic efficiency of 0.027 µM⁻¹min⁻¹ in the reconstituted system. On the basis of this finding, we were able to biosynthesise the conjugate of clopidogrel with 3-nitropyridine-2-thiol, referred to as clopNPT, and examined its antiplatelet activity in male New Zealand white rabbits. After administration as intravenous bolus at 2 mg/kg, the clopNPT conjugate was rapidly converted to the AM leading to the inhibition of platelet aggregation (IPA). Analyses of the blood samples drawn at various time points showed that intravenous administration of clopNPT led to ~70% IPA within 1 hour and the IPA persisted for more than 3 hours. Since the antiplatelet activity of clopNPT does not require bioactivation by P450s, the mixed disulfide conjugate of clopidogrel has the potential to overcome the inter-individual variability in clopidogrel therapy. PMID:25230737

  14. Disulfide scrambling in superoxide dismutase 1 reduces its cytotoxic effect in cultured cells and promotes protein aggregation.

    Directory of Open Access Journals (Sweden)

    Lina Leinartaitė

    Full Text Available Mutations in the gene coding for superoxide dismutase 1 (SOD1 are associated with familiar forms of the neurodegenerative disease amyotrophic lateral sclerosis (ALS. These mutations are believed to result in a "gain of toxic function", leading to neuronal degeneration. The exact mechanism is still unknown, but misfolding/aggregation events are generally acknowledged as important pathological events in this process. Recently, we observed that demetallated apoSOD1, with cysteine 6 and 111 substituted for alanine, is toxic to cultured neuroblastoma cells. This toxicity depended on an intact, high affinity Zn(2+ site. It was therefor contradictory to discover that wild-type apoSOD1 was not toxic, despite of its high affinity for Zn(2+. This inconsistency was hypothesized to originate from erroneous disulfide formation involving C6 and C111. Using high resolution non-reducing SDS-PAGE, we have in this study demonstrated that the inability of wild-type apoSOD1 to cause cell death stems from formation of non-native intra-molecular disulfides. Moreover, monomeric apoSOD1 variants capable of such disulfide scrambling aggregated into ThT positive oligomers under physiological conditions without agitation. The oligomers were stabilized by inter-molecular disulfides and morphologically resembled what has in other neurodegenerative diseases been termed protofibrils. Disulfide scrambling thus appears to be an important event for misfolding and aggregation of SOD1, but may also be significant for protein function involving cysteines, e.g. mitochondrial import and copper loading.

  15. Conformational difference in human IgG2 disulfide isoforms revealed by hydrogen/deuterium exchange mass spectrometry.

    Science.gov (United States)

    Zhang, Aming; Fang, Jing; Chou, Robert Y-T; Bondarenko, Pavel V; Zhang, Zhongqi

    2015-03-17

    Both recombinant and natural human IgG2 antibodies have several different disulfide bond isoforms, which possess different global structures, thermal stabilities, and biological activities. A detailed mapping of the structural difference among IgG2 disulfide isoforms, however, has not been established. In this work, we employed hydrogen/deuterium exchange mass spectrometry to study the conformation of three major IgG2 disulfide isoforms known as IgG2-B, IgG2-A1, and IgG2-A2 in two recombinant human IgG2 monoclonal antibodies. By comparing the protection factors between amino acid residues in isoforms B and A1 (the classical form), we successfully identified several local regions in which the IgG2-B isoform showed more solvent protection than the IgG2-A1 isoform. On the basis of three-dimensional structural models of IgG2, these identified regions were located on the Fab domains, close to the hinge, centered on the side where the two Fab arms faced each other in spatial proximity. We speculated that in the more solvent-protected B isoform, the two Fab arms were brought into contact by the nonclassical disulfide bonds, resulting in a more compact global structure. Loss of Fab domain flexibility in IgG2-B could limit its ability to access cell-surface epitopes, leading to reduced antigen binding potency. The A2 isoform was previously found to have disulfide linkages similar to those of the classical A1 isoform, but with different biophysical behaviors. Our data indicated that, compared to IgG2-A1, IgG2-A2 had less solvent protection in some heavy-chain Fab regions close the hinge, suggesting that the A2 isoform had more flexible Fab domains. PMID:25730439

  16. Analysis of glutathione and glutathione disulfide in whole cells and mitochondria by postcolumn derivatization high-performance liquid chromatography with ortho-phthalaldehyde.

    Science.gov (United States)

    Lenton, K J; Therriault, H; Wagner, J R

    1999-10-01

    A method is described for the detection of glutathione (GSH) and glutathione disulfide (GSSG) based on a HPLC postcolumn reaction with ortho-phthalaldehyde (OPT) at pH 12 followed by fluorescence detection. Although similar methods have been reported, the high pH of the postcolumn reaction adds considerable selectivity and sensitivity to the measurement of GSH and glutathione disulfide. The limit of detection approaches 100 fmol, which is sufficient to detect whole-cell glutathione disulfide in 10,000 cells or mitochondrial glutathione disulfide in 20 million cells. Using this method, glutathione and glutathione disulfide were measured in human lymphocytes, granulocytes, and cultured Jurkat T cells, as well as in the corresponding samples of mitochondria. The percentage of glutathione disulfide to total glutathione in whole-cell extracts was approximately 1%. In contrast, the percentage was relatively high in mitochondria, with the mitochondria of granulocytes having the highest (25%) followed by those of lymphocytes (15%) and finally by cultured Jurkat T cells (9%). This method extends the analysis of glutathione and glutathione disulfide to mitochondria obtained from a relatively small number of cells. PMID:10527505

  17. Production of recombinant disulfide-rich venom peptides for structural and functional analysis via expression in the periplasm of E. coli.

    Directory of Open Access Journals (Sweden)

    Julie K Klint

    Full Text Available Disulfide-rich peptides are the dominant component of most animal venoms. These peptides have received much attention as leads for the development of novel therapeutic agents and bioinsecticides because they target a wide range of neuronal receptors and ion channels with a high degree of potency and selectivity. In addition, their rigid disulfide framework makes them particularly well suited for addressing the crucial issue of in vivo stability. Structural and functional characterization of these peptides necessitates the development of a robust, reliable expression system that maintains their native disulfide framework. The bacterium Escherichia coli has long been used for economical production of recombinant proteins. However, the expression of functional disulfide-rich proteins in the reducing environment of the E. coli cytoplasm presents a significant challenge. Thus, we present here an optimised protocol for the expression of disulfide-rich venom peptides in the periplasm of E. coli, which is where the endogenous machinery for production of disulfide-bonds is located. The parameters that have been investigated include choice of media, induction conditions, lysis methods, methods of fusion protein and peptide purification, and sample preparation for NMR studies. After each section a recommendation is made for conditions to use. We demonstrate the use of this method for the production of venom peptides ranging in size from 2 to 8 kDa and containing 2-6 disulfide bonds.

  18. Molecular dynamics simulations on pars intercerebralis major peptide-C (PMP-C) reveal the role of glycosylation and disulfide bonds in its enhanced structural stability and function.

    Science.gov (United States)

    Kaushik, Sandeep; Mohanty, Debasisa; Surolia, Avadhesha

    2012-01-01

    Fucosylation of Thr 9 in pars intercerebralis major peptide-C (PMP-C) enhances its structural stability and functional ability as a serine protease inhibitor. In order to understand the role of disulfide bonds and glycosylation on the structure and function of PMP-C, we have carried out multiple explicit solvent molecular dynamics (MD) simulations on fucosylated and non-fucosylated forms of PMP-C, both in the presence and absence of the disulfide bonds. Our simulations revealed that there were no significant structural changes in the native disulfide bonded forms of PMP-C due to fucosylation. On the other hand, the non-fucosylated form of PMP-C without disulfide bonds showed larger deviations from the starting structure than the fucosylated form. However, the structural deviations were restricted to the terminal regions while core β-sheet retained its hydrogen bonded structure even in absence of disulfide bonds as well as fucosylation. Interestingly, fucosylation of disulfide bonded native PMP-C led to a decreased thermal flexibility in the residue stretch 29-32 which is known to interact with the active site of the target proteases. Our analysis revealed that disulfide bonds covalently connect the residue stretch 29-32 to the central β-sheet of PMP-C and using a novel network of side chain interactions and disulfide bonds fucosylation at Thr 9 is altering the flexibility of the stretch 29-32 located at a distal site. Thus, our simulations explain for the first time, how presence of disulfide bonds between conserved cysteines and fucosylation enhance the function of PMP-C as a protease inhibitor.

  19. Comprehensive identification of disulfide bonds using non-specific proteinase K digestion and CID-cleavable crosslinking analysis methodology for Orbitrap LC/ESI-MS/MS data.

    Science.gov (United States)

    Makepeace, Karl A T; Serpa, Jason J; Petrotchenko, Evgeniy V; Borchers, Christoph H

    2015-11-01

    Disulfide bonds are valuable constraints in protein structure modeling. The Cys-Cys disulfide bond undergoes specific fragmentation under CID and, therefore, can be considered as a CID-cleavable crosslink. We have recently reported on the benefits of using non-specific digestion with proteinase K for inter-peptide crosslink determination. Here, we describe an updated application of our CID-cleavable crosslink analysis software and our crosslinking analysis with non-specific digestion methodology for the robust and comprehensive determination of disulfide bonds in proteins, using Orbitrap LC/ESI-MS/MS data.

  20. Probing the Conformational and Functional Consequences of Disulfide Bond Engineering in Growth Hormone by Hydrogen-Deuterium Exchange Mass Spectrometry Coupled to Electron Transfer Dissociation

    DEFF Research Database (Denmark)

    Seger, Signe T; Breinholt, Jens; Faber, Johan H;

    2015-01-01

    spectrometry (HDX-MS) to map the impact of the new disulfide bond on the conformational dynamics of this new hGH variant. Compared to wild type hGH, the variant exhibits reduced loop dynamics, indicating a stabilizing effect of the introduced disulfide bond. Furthermore, the disulfide bond exhibits longer...... ranging effects, stabilizing a short α-helix quite distant from the mutation sites, but also rendering a part of the α-helical hGH core slightly more dynamic. In the regions where the hGH variant exhibits a different deuterium uptake than the wild type protein, electron transfer dissociation (ETD...

  1. A novel disulfide-rich protein motif from avian eggshell membranes.

    Directory of Open Access Journals (Sweden)

    Vamsi K Kodali

    Full Text Available Under the shell of a chicken egg are two opposed proteinaceous disulfide-rich membranes. They are fabricated in the avian oviduct using fibers formed from proteins that are extensively coupled by irreversible lysine-derived crosslinks. The intractability of these eggshell membranes (ESM has slowed their characterization and their protein composition remains uncertain. In this work, reductive alkylation of ESM followed by proteolytic digestion led to the identification of a cysteine rich ESM protein (abbreviated CREMP that was similar to spore coat protein SP75 from cellular slime molds. Analysis of the cysteine repeats in partial sequences of CREMP reveals runs of remarkably repetitive patterns. Module a contains a C-X(4-C-X(5-C-X(8-C-X(6 pattern (where X represents intervening non-cysteine residues. These inter-cysteine amino acid residues are also strikingly conserved. The evolutionarily-related module b has the same cysteine spacing as a, but has 11 amino acid residues at its C-terminus. Different stretches of CREMP sequences in chicken genomic DNA fragments show diverse repeat patterns: e.g. all a modules; an alternation of a-b modules; or an a-b-b arrangement. Comparable CREMP proteins are found in contigs of the zebra finch (Taeniopygia guttata and in the oviparous green anole lizard (Anolis carolinensis. In all these cases the long runs of highly conserved modular repeats have evidently led to difficulties in the assembly of full length DNA sequences. Hence the number, and the amino acid lengths, of CREMP proteins are currently unknown. A 118 amino acid fragment (representing an a-b-a-b pattern from a chicken oviduct EST library expressed in Escherichia coli is a well folded, highly anisotropic, protein with a large chemical shift dispersion in 2D solution NMR spectra. Structure is completely lost on reduction of the 8 disulfide bonds of this protein fragment. Finally, solid state NMR spectra suggest a surprising degree of order in intact

  2. Layer-modulated synthesis of uniform tungsten disulfide nanosheet using gas-phase precursors

    Science.gov (United States)

    Park, Jusang; Lee, Wonseon; Choi, Taejin; Hwang, Sung-Hwan; Myoung, Jae Min; Jung, Jae-Hoon; Kim, Soo-Hyun; Kim, Hyungjun

    2015-01-01

    The synthesis of layered transition-metal-disulfide (MS2, M = Mo, W) nanosheets with layer controllability and large-area uniformity is an essential requirement for their application in electronic and optical devices. In this report, we describe a synthesis process of WS2 nanosheets with layer controllability and high uniformity using chemical vapor deposition (CVD) and WCl6 and H2S as gas-phase precursors. Through this process, we can systematically modulate the thickness of WS2 nanosheets by controlling the duration of the reaction between WCl6 and H2S. The CVD-grown WS2 nanosheets exhibit good stoichiometry as well as dependencies of a clear Raman shift and bandgap on the number of layers. These properties are confirmed by X-ray photoemission spectroscopy, Raman spectroscopy, and photoluminescence measurements. The number of layers of WS2 nanosheets is confirmed by atomic force microscopy. Finally, we demonstrate the fabrication and performance of a photodetector based on a hybrid structure consisting of graphene and a WS2 nanosheet.The synthesis of layered transition-metal-disulfide (MS2, M = Mo, W) nanosheets with layer controllability and large-area uniformity is an essential requirement for their application in electronic and optical devices. In this report, we describe a synthesis process of WS2 nanosheets with layer controllability and high uniformity using chemical vapor deposition (CVD) and WCl6 and H2S as gas-phase precursors. Through this process, we can systematically modulate the thickness of WS2 nanosheets by controlling the duration of the reaction between WCl6 and H2S. The CVD-grown WS2 nanosheets exhibit good stoichiometry as well as dependencies of a clear Raman shift and bandgap on the number of layers. These properties are confirmed by X-ray photoemission spectroscopy, Raman spectroscopy, and photoluminescence measurements. The number of layers of WS2 nanosheets is confirmed by atomic force microscopy. Finally, we demonstrate the fabrication

  3. Hydrothermal Synthesis of Disulfide-Containing Uranyl Compounds. In Situ Ligand Synthesis versus Direct Assembly

    Energy Technology Data Exchange (ETDEWEB)

    Rowland, Clare E. [George Washington Univ., Washington, DC (United States); Belai, Nebebech [George Washington Univ., Washington, DC (United States); Knope, Karah E. [George Washington Univ., Washington, DC (United States); Cahill, Christopher L. [George Washington Univ., Washington, DC (United States)

    2010-01-29

    Three disulfide-containing uranyl compounds, [UO2(C7H4O2S)3]·H2O (1), [UO2(C7H4O2S)2(C7H5O2S)] (2), and [UO2(C7H4O2S)4] (3) have been hydrothermally synthesized. Both in situ disulfide bond formation from 3- and 4-mercaptobenzoic acid (C7H5O2S, MBA) to yield 3,3'- and 4,4'-dithiobisbenzoic acid (C14H8O4S2, DTBA) and direct assembly with the presynthesized dimeric ligands have been explored. While the starting materials 4-MBA and 4,4'-DTBA both yield 2 via in situ ligand synthesis and direct assembly, respectively, we observe the formation of 1 from the starting material 3-MBA via in situ ligand synthesis and of 3 from the direct assembly of the uranyl cation with 3,3'-DTBA. Concurrently with the synthesis of 1 and 2, we have observed the in situ formation of the crystalline dimeric organic species, 3,3'-DTBA, [(C7H5O2S)2] (4) and 4,4'-DTBA, [(C7H5O2S)2] (5). Herein we report the synthesis and crystallographic characterization of 1-5, as well as observations regarding the utility of product formation via direct assembly and in situ ligand synthesis.

  4. Dissulfeto de molibdênio, um material multifuncional e surpreendente Molybdenum disulfide, a multifunctional and remarkable material

    Directory of Open Access Journals (Sweden)

    Fernando Wypych

    2002-02-01

    Full Text Available The aim of this work is to review the chemical and physical properties of layered molybdenum disulfide. The three polymorphic/polytypic modifications of the compound were found, the polytypes 2H (molybdenite and 3R are semiconductors while the polymorph 1T is an electronic conductor. 2H-MoS2 has several important industrial applications as hydrotreatment catalysts, energy storage devices, solar cells, solid lubricants, among others. When intercalated, the 2H phase changes to a distorted 1T phase, producing unstable intercalation compounds that can be exfoliated in solution, producing single layers and consequently nanocomposites. The direct synthesis of the 1T phase produces stable intercalation compounds. Recently molybdenum disulfide was prepared as nanotubes and fulerene-like structures that bring new insights in the investigation of this important material.

  5. Engineering a disulfide bond in the lid hinge region of Rhizopus chinensis lipase: increased thermostability and altered acyl chain length specificity.

    Directory of Open Access Journals (Sweden)

    Xiao-Wei Yu

    Full Text Available The key to enzyme function is the maintenance of an appropriate balance between molecular stability and structural flexibility. The lid domain which is very important for "interfacial activation" is the most flexible part in the lipase structure. In this work, rational design was applied to explore the relationship between lid rigidity and lipase activity by introducing a disulfide bond in the hinge region of the lid, in the hope of improving the thermostability of R. chinensis lipase through stabilization of the lid domain without interfering with its catalytic performance. A disulfide bridge between F95C and F214C was introduced into the lipase from R. chinensis in the hinge region of the lid according to the prediction of the "Disulfide by Design" algorithm. The disulfide variant showed substantially improved thermostability with an eleven-fold increase in the t(1/2 value at 60°C and a 7°C increase of T(m compared with the parent enzyme, probably contributed by the stabilization of the geometric structure of the lid region. The additional disulfide bond did not interfere with the catalytic rate (k(cat and the catalytic efficiency towards the short-chain fatty acid substrate, however, the catalytic efficiency of the disulfide variant towards pNPP decreased by 1.5-fold probably due to the block of the hydrophobic substrate channel by the disulfide bond. Furthermore, in the synthesis of fatty acid methyl esters, the maximum conversion rate by RCLCYS reached 95% which was 9% higher than that by RCL. This is the first report on improving the thermostability of the lipase from R. chinensis by introduction of a disulfide bond in the lid hinge region without compromising the catalytic rate.

  6. Protein and non-protein sulfhydryls and disulfides in gastric mucosa and liver after gastrotoxic chemicals and sucralfate: Possible new targets of pharmacologic agents

    Institute of Scientific and Technical Information of China (English)

    Lajos Nagy; Miki Nagata; Sandor Szabo

    2007-01-01

    AIM: To investigate the role of major non-protein and protein sulfhydryls and disulfides in chemically induced gastric hemorrhagic mucosal lesions (HML) and the mechanism of gastroprotective effect of sucralfate.METHODS: Rats were given 1 mL of 75% ethanol, 25%NaCl, 0.6 mol/L HCI, 0.2 mol/L NaOH or 1% ammonia solutions intragastrically (i.g.) and sacrificed 1, 3, 6 or 12 min later. Total (reduced and oxidized) glutathione (GSH + GSSG), glutathione disulfide (GSSG), protein free sulfhydryls (PSH), protein-glutathione mixed disulfides (PSSG) and protein cystine disulfides (PSSP) were measured in gastric mucosa and liver.RESULTS: Reduced glutathione (GSH) was depleted in the gastric mucosa after ethanol, HCI or NaCl exposure,while oxidized glutathione (GSSG) concentrations increased, except by HCI and NaOH exposure. Decreased levels of PSH after exposure to ethanol were observed,NaCl or NaOH while the total protein disulfides were increased. Ratios of reduced to oxidized glutathione or sulfhydrils to disulfides were decreased by all chemicals.No changes in thiol homeostasis were detected in the liver after i.g. abbreviation should be spelled out the first time here administration of ethanol. Sucralfate increased the concentrations of GSH and PSH and prevented the ethanol-induced changes in gastric mucosal thiol concentrations.CONCLUSION: Our modified methods are now suitable for direct measurements of major protein and nonprotein thiols/disulfides in the gastric mucosa or liver.A common element in the pathogenesis of chemically induced HML and in the mechanism of gastroprotective drugs seems to be the decreased ratios of reduced and oxidized glutathione as well as protein sulfhydryls and disulfides.

  7. Thioredoxin Reductase Is Essential for Thiol/Disulfide Redox Control and Oxidative Stress Survival of the Anaerobe Bacteroides fragilis▿ †

    OpenAIRE

    Rocha, Edson R.; Tzianabos, Arthur O; Smith, C. Jeffrey

    2007-01-01

    Results of this study showed that the anaerobic, opportunistic pathogen Bacteroides fragilis lacks the glutathione/glutaredoxin redox system and possesses an extensive number of putative thioredoxin (Trx) orthologs. Analysis of the genome sequence revealed six Trx orthologs and an absence of genes required for synthesis of glutathione and glutaredoxins. In addition, it was shown that the thioredoxin reductase (TrxB)/Trx system is the major or sole redox system for thiol/disulfide cellular hom...

  8. Effect of trastuzumab interchain disulfide bond cleavage on Fcγ receptor binding and antibody-dependent tumour cell phagocytosis.

    Science.gov (United States)

    Suzuki, Mami; Yamanoi, Ayaka; Machino, Yusuke; Ootsubo, Michiko; Izawa, Ken-ichi; Kohroki, Junya; Masuho, Yasuhiko

    2016-01-01

    The Fc domain of human IgG1 binds to Fcγ receptors (FcγRs) to induce effector functions such as phagocytosis. There are four interchain disulfide bonds between the H and L chains. In this study, the disulfide bonds within the IgG1 trastuzumab (TRA), which is specific for HER2, were cleaved by mild S-sulfonation or by mild reduction followed by S-alkylation with three different reagents. The cleavage did not change the binding activities of TRA to HER2-bearing SK-BR-3 cells. The binding activities of TRA to FcγRIIA and FcγRIIB were greatly enhanced by modification with mild reduction and S-alkylation with ICH2CONH2 or N-(4-aminophenyl) maleimide, while the binding activities of TRA to FcγRI and FcγRIIIA were decreased by any of the four modifications. However, the interchain disulfide bond cleavage by the different modifications did not change the antibody-dependent cell-mediated phagocytosis (ADCP) of SK-BR-3 cells by activated THP-1 cells. The order of FcγR expression levels on the THP-1 cells was FcγRII > FcγRI > FcγRIII and ADCP was inhibited by blocking antibodies against FcγRI and FcγRII. These results imply that the effect of the interchain disulfide bond cleavage on FcγRs binding and ADCP is dependent on modifications of the cysteine residues and the FcγR isotypes. PMID:26254483

  9. Protein disulfide isomerase acts as an injury response signal that enhances fibrin generation via tissue factor activation

    OpenAIRE

    Reinhardt, Christoph; von Brühl, Marie-Luise; Manukyan, Davit; Grahl, Lenka; Lorenz, Michael; Altmann, Berid; Dlugai, Silke; Hess, Sonja; Konrad, Ildiko; Orschiedt, Lena; Mackman, Nigel; Ruddock, Lloyd; Massberg, Steffen; Engelmann, Bernd

    2008-01-01

    The activation of initiator protein tissue factor (TF) is likely to be a crucial step in the blood coagulation process, which leads to fibrin formation. The stimuli responsible for inducing TF activation are largely undefined. Here we show that the oxidoreductase protein disulfide isomerase (PDI) directly promotes TF-dependent fibrin production during thrombus formation in vivo. After endothelial denudation of mouse carotid arteries, PDI was released at the injury site from adherent platelets...

  10. Discovery of a linear cyclotide from the bracelet subfamily and its disulfide mapping by top-down mass spectrometry.

    Science.gov (United States)

    Nguyen, Giang Kien Truc; Zhang, Sen; Wang, Wei; Wong, Clarence Tsun Ting; Nguyen, Ngan Thi Kim; Tam, James P

    2011-12-30

    Cyclotides are heat-stable macrocyclic peptides from plants that display a wide range of biological activities. They can be divided into two subfamilies: Möbius or bracelet, based on the presence or absence of a cis-proline residue in loop 5, respectively. Currently, over 150 cyclotides have been discovered, but only four linear variants of the Möbius subfamily have been hitherto isolated. In this study, we report the discovery of two novel cyclotides, hedyotide B1 and hedyotide B2, from the aerial parts of Hedyotis biflora. Hedyotide B1 has a cyclic cystine knot structure typical of cyclotides. Interestingly, hedyotide B2 possesses a linear backbone and is the first linear representative of the bracelet subfamily. Disulfide mapping of hedyotide B2 by a top-down MS/MS approach showed that it shares the same knotted disulfide arrangement as conventional cyclotides. Its unfolding pathway also showed that the penetrating disulfide bond Cys III-VI is the most stable disulfide linkage. Cloning of the gene encoding hedyotide B2 revealed a nonsense mutation that introduces a premature stop codon at the conserved Asn residue position, which is essential for an end-to-end backbone ligation. Biophysical characterization showed that hedyotide B2 was more susceptible to exopeptidase degradation as compared with hedyotide B1. Hedyotide B2 was also inactive against all four tested bacterial strains, whereas hedyotide B1 was bactericidal to Escherichia coli and Streptococcus salivarius at low micromolar concentration. Our results provide a deeper understanding of the structures, functions, and biosynthetic processing of cyclotides and uncyclotides in plants. PMID:21979955

  11. The effect of disulfide bond introduction and related Cys/Ser mutations on the stability of a cyclohexanone monooxygenase.

    Science.gov (United States)

    Schmidt, Sandy; Genz, Maika; Balke, Kathleen; Bornscheuer, Uwe T

    2015-11-20

    Baeyer-Villiger monooxygenases (BVMO) belong to the class B of flavin-dependent monooxygenases (type I BVMOs) and catalyze the oxidation of (cyclic) ketones into esters and lactones. The prototype BVMO is the cyclohexanone monooxygenase (CHMO) from Acinetobacter sp. NCIMB 9871. This enzyme shows an impressive substrate scope with a high chemo-, regio- and/or enantioselectivity. BVMO reactions are often difficult, if not impossible to achieve by chemical approaches and this makes these enzymes thus highly desired candidates for industrial applications. Unfortunately, the industrial use is hampered by several factors related to the lack of stability of these biocatalysts. Thus, the aim of this study was to improve the CHMO's long-term stability, one of the most relevant parameter for biocatalytic processes, and additionally its stability against oxidation. We used an easy computational method for the prediction of stabilizing disulfide bonds in the CHMO-scaffold. The three most promising predicted disulfide pairs were created and biochemically characterized. The most oxidatively stable variant (Y411C-A463C) retained nearly 60% activity after incubation with 25 mM H2O2 whereas the wild type retained only 16%. In addition, one extra disulfide pair (T415C-A463C) was created and tested for increased stability. The melting temperature (Tm) of this variant was increased by 5°C with simultaneous improved long-term stability. After verification by ABD-F labeling that this mutant does not form a disulfide bond, single and double Cys/Ser mutants were prepared and investigated. Subsequent analysis revealed that the T415C single point variant is the most stable variant with a 30-fold increased long-term stability (33% residual activity after 24h incubation at 25°C) showcasing a great achievement for practical applications.

  12. Redox Reactivity of Cerium Oxide Nanoparticles Induces the Formation of Disulfide Bridges in Thiol-Containing Biomolecules.

    Science.gov (United States)

    Rollin-Genetet, Françoise; Seidel, Caroline; Artells, Ester; Auffan, Mélanie; Thiéry, Alain; Vidaud, Claude

    2015-12-21

    The redox state of disulfide bonds is implicated in many redox control systems, such as the cysteine-cystine couple. Among proteins, ubiquitous cysteine-rich metallothioneins possess thiolate metal binding groups susceptible to metal exchange in detoxification processes. CeO2 NPs are commonly used in various industrial applications due to their redox properties. These redox properties that enable dual oxidation states (Ce(IV)/Ce(III)) to exist at their surface may act as oxidants for biomolecules. The interaction among metallothioneins, cysteine, and CeO2 NPs was investigated through various biophysical approaches to shed light on the potential effects of the Ce(4+)/Ce(3+) redox system on the thiol groups of these biomolecules. The possible reaction mechanisms include the formation of a disulfide bridge/Ce(III) complex resulting from the interaction between Ce(IV) and the thiol groups, leading to metal unloading from the MTs, depending on their metal content and cluster type. The formation of stable Ce(3+) disulfide complexes has been demonstrated via their fluorescence properties. This work provides the first evidence of thiol concentration-dependent catalytic oxidation mechanisms between pristine CeO2 NPs and thiol-containing biomolecules. PMID:26566067

  13. Design, Synthesis and Biological Evaluation of Brain-Targeted Thiamine Disulfide Prodrugs of Ampakine Compound LCX001

    Directory of Open Access Journals (Sweden)

    Dian Xiao

    2016-04-01

    Full Text Available Ampakine compounds have been shown to reverse opiate-induced respiratory depression by activation of amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA glutamate receptors. However, their pharmacological exploitations are hindered by low blood-brain barrier (BBB permeability and limited brain distribution. Here, we explored whether thiamine disulfide prodrugs with the ability of “lock-in” can be used to solve these problems. A series of thiamine disulfide prodrugs 7a–7f of ampakine compound LCX001 was synthesized and evaluated. The trials in vitro showed that prodrugs 7e, 7d, 7f possessed a certain stability in plasma and quickly decomposed in brain homogenate by the disulfide reductase. In vivo, prodrug 7e decreased the peripheral distribution of LCX001 and significantly increased brain distribution of LCX001 after i.v. administration. This compound showed 2.23- and 3.29-fold greater increases in the AUC0-t and MRT0-t of LCX001 in brain, respectively, than did LCX001 itself. A preliminary pharmacodynamic study indicated that the required molar dose of prodrug 7e was only one eighth that of LCX001 required to achieve the same effect in mice. These findings provide an important reference to evaluate the clinical outlook of ampakine compounds.

  14. Phosphorothioate anti-sense oligonucleotides: the kinetics and mechanism of the generation of the sulfurising agent from phenylacetyl disulfide (PADS).

    Science.gov (United States)

    Scotson, James L; Andrews, Benjamin I; Laws, Andrew P; Page, Michael I

    2016-09-21

    The synthesis of phosphorothioate oligonucleotides is often accomplished in the pharmaceutical industry by the sulfurisation of the nucleotide-phosphite using phenylacetyl disulfide (PADS) which has an optimal combination of properties. This is best achieved by an initial 'ageing' of PADS for 48 h in acetonitrile with 3-picoline to generate polysulfides. The initial base-catalysed degradation of PADS occurs by an E1cB-type elimination to generate a ketene and acyldisulfide anion. Proton abstraction to reversibly generate a carbanion is demonstrated by H/D exchange, the rate of which is greatly increased by electron-withdrawing substituents in the aromatic ring of PADS. The ketene can be trapped intramolecularly by an o-allyl group. The disulfide anion generated subsequently attacks unreacted PADS on sulfur to give polysulfides, the active sulfurising agent. The rate of degradation of PADS is decreased by less basic substituted pyridines and is only first order in PADS indicating that the rate-limiting step is formation of the disulfide anion from the carbanion. PMID:27531007

  15. Quantitation of protein S-glutathionylation by liquid chromatography-tandem mass spectrometry: correction for contaminating glutathione and glutathione disulfide.

    Science.gov (United States)

    Bukowski, Michael R; Bucklin, Christopher; Picklo, Matthew J

    2015-01-15

    Protein S-glutathionylation is a posttranslational modification that links oxidative stimuli to reversible changes in cellular function. Protein-glutathione mixed disulfide (PSSG) is commonly quantified by reduction of the disulfide and detection of the resultant glutathione species. This methodology is susceptible to contamination by free unreacted cellular glutathione (GSH) species, which are present in 1000-fold greater concentration. A liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based method was developed for quantification of glutathione and glutathione disulfide (GSSG), which was used for the determination of PSSG in biological samples. Analysis of rat liver samples demonstrated that GSH and GSSG coprecipitated with proteins similar to the range for PSSG in the sample. The use of [(13)C2,(5)N]GSH and [(13)C4,(5)N2]GSSG validated these results and demonstrated that the release of GSH from PSSG did not occur during sample preparation and analysis. These data demonstrate that GSH and GSSG contamination must be accounted for when determining PSSG content in cellular/tissue preparations. A protocol for rinsing samples to remove the adventitious glutathione species is demonstrated. The fragmentation patterns for glutathione were determined by high-resolution mass spectrometry, and candidate ions for detection of PSSG on protein and protein fragments were identified.

  16. A role for sperm surface protein disulfide isomerase activity in gamete fusion: evidence for the participation of ERp57.

    Science.gov (United States)

    Ellerman, Diego A; Myles, Diana G; Primakoff, Paul

    2006-06-01

    In mammals, sperm-egg interaction is based on molecular events either unique to gametes or also present in somatic cells. In gamete fusion, it is unknown which features are gamete specific and which are shared with other systems. Conformational changes mediated by thiol-disulfide exchange are involved in the activation of some virus membrane fusion proteins. Here we asked whether that mechanism is also operative in sperm-egg fusion. Different inhibitors of protein disulfide isomerase (PDI) activity were able to inhibit sperm-egg fusion in vitro. While pretreatment of oocytes had no effect, pretreatment of sperm reduced their fusion ability. Some members of the PDI family were detected on the sperm head, and use of specific antibodies and substrates suggested that the oxidoreductase ERp57 has a role in gamete fusion. The results support the idea that thiol-disulfide exchange is a mechanism that may act in gamete fusion to produce conformational changes in fusion-active proteins. PMID:16740484

  17. Disulfide bond-dependent mechanism of protection against oxidative stress in pyruvate-ferredoxin oxidoreductase of anaerobic Desulfovibrio bacteria.

    Science.gov (United States)

    Vita, Nicolas; Hatchikian, E Claude; Nouailler, Matthieu; Dolla, Alain; Pieulle, Laetitia

    2008-01-22

    Oxidative decarboxylation of pyruvate forming acetyl-coenzyme A is a crucial step in many metabolic pathways. In most anaerobes, this reaction is carried out by pyruvate-ferredoxin oxidoreductase (PFOR), an enzyme normally oxygen sensitive except in Desulfovibrio africanus (Da), where it shows an abnormally high oxygen stability. Using site-directed mutagenesis, we have specified a disulfide bond-dependent protective mechanism against oxidative conditions in Da PFOR. Our data demonstrated that the two cysteine residues forming the only disulfide bond in the as-isolated PFOR are crucial for the stability of the enzyme in oxidative conditions. A methionine residue located in the environment of the proximal [4Fe-4S] cluster was also found to be essential for this protective mechanism. In vivo analysis demonstrated unambiguously that PFOR in Da cells as well as two other Desulfovibrio species was efficiently protected against oxidative stress. Importantly, a less active but stable Da PFOR in oxidized cells rapidly reactivated when returned to anaerobic medium. Our work demonstrates the existence of an elegant disulfide bond-dependent reversible mechanism, found in the Desulfovibrio species to protect one of the key enzymes implicated in the central metabolism of these strict anaerobes. This new mechanism could be considered as an adaptation strategy used by sulfate-reducing bacteria to cope with temporary oxidative conditions and to maintain an active dormancy. PMID:18161989

  18. Fast and Efficient non-reduced Lys-C digest using pressure cycling technology for antibody disulfide mapping by LC-MS.

    Science.gov (United States)

    Cheng, Ying; Chen, Yonghong; Yu, Christopher

    2016-09-10

    Conventional sample preparation for antibody disulfide mapping often requires relatively long digestion time (from several hours to overnight) and relatively high endoproteinase concentration. These conditions are typically necessitated by the fact that antibody molecules are not sufficiently denatured under non-reduced conditions and chaotropic agents are used during digestion to achieve optimal denaturation. Disulfide scrambling can occur as artifacts of digestion as proteins are incubated for extended periods, often at neutral to slightly alkaline pH conditions. Shortening digestion time and lowering the pH during digestion frequently result in incomplete peptide cleavages or variable recoveries. Here, we report the development of a fast and efficient non-reduced Lys-C digestion method based on pressure cycling technology (PCT) and its application in determining disulfide-linkages in monoclonal antibodies (mAbs). Conditions were optimized to ensure complete digestion of the mAb with minimal sample preparation-related disulfide scrambling. The PCT-based method was able to generate up to 10-fold signal increase for some disulfide peptides in a 1h Lys-C digestion compared to the conventional bench-top digestion method. As a result of the shorter digestion time, disulfide scrambling that is seen as a major assay artifact of the conventional method was reduced to less than 0.05% in tested molecules. The results show that the PCT-based method offers fast digestion in a shorter time for all the mAbs tested. PMID:27429370

  19. Investigation of some Schiff base compounds containing disulfide bond as HCl corrosion inhibitors for mild steel

    Energy Technology Data Exchange (ETDEWEB)

    Behpour, M., E-mail: m.behpour@kashanu.ac.i [Department of Chemistry, Faculty of Science, University of Kashan, Kashan (Iran, Islamic Republic of); Ghoreishi, S.M.; Mohammadi, N. [Department of Chemistry, Faculty of Science, University of Kashan, Kashan (Iran, Islamic Republic of); Soltani, N. [Payame Noor University (PNU), Shahin Shahr Branch, Isfahan (Iran, Islamic Republic of); Salavati-Niasari, M. [Department of Chemistry, Faculty of Science, University of Kashan, Kashan (Iran, Islamic Republic of)

    2010-12-15

    Research highlights: {yields} All studied Schiff bases are effective inhibitors for mild steel in 2.0 M HCl. {yields} The inhibition is accomplished by adsorption of molecules on the steel surface. {yields} Examined Schiff bases behave as mixed type inhibitor. {yields} Feed back bonds form between the Schiff bases and steel surface. {yields} Quantum chemical calculations were applied to explain the experimental results. - Abstract: The inhibition performance of three Schiff bases containing disulfide bond as corrosion inhibitors for mild steel in 2.0 M HCl has been investigated by weight loss measurements, potentiodynamic polarization measurements and electrochemical impedance spectroscopy (EIS). Potentiodynamic polarization study showed that all the inhibitors are mixed type. The adsorption of inhibitors on mild steel surface was found to follow Langmuir adsorption isotherm and the adsorption isotherm parameters (K{sub ads}, {Delta}G{sub ads}) were determined. Quantum chemical calculations were further applied to reveal the adsorption structure and explain the experimental results. Some samples of mild steel were examined by SEM.

  20. Utilizing self-assembled-monolayer-based gate dielectrics to fabricate molybdenum disulfide field-effect transistors

    Science.gov (United States)

    Kawanago, Takamasa; Oda, Shunri

    2016-01-01

    In this study, we apply self-assembled-monolayer (SAM)-based gate dielectrics to the fabrication of molybdenum disulfide (MoS2) field-effect transistors. A simple fabrication process involving the selective formation of a SAM on metal oxides in conjunction with the dry transfer of MoS2 flakes was established. A subthreshold slope (SS) of 69 mV/dec and no hysteresis were demonstrated with the ultrathin SAM-based gate dielectrics accompanied by a low gate leakage current. The small SS and no hysteresis indicate the superior interfacial properties of the MoS2/SAM structure. Cross-sectional transmission electron microscopy revealed a sharp and abrupt interface of the MoS2/SAM structure. The SAM-based gate dielectrics are found to be applicable to the fabrication of low-voltage MoS2 field-effect transistors and can also be extended to various layered semiconductor materials. This study opens up intriguing possibilities of SAM-based gate dielectrics in functional electronic devices.

  1. Ammonia gas sensors based on poly (3-hexylthiophene)-molybdenum disulfide film transistors

    Science.gov (United States)

    Xie, Tao; Xie, Guangzhong; Su, Yuanjie; Hongfei, Du; Ye, Zongbiao; Jiang, Yadong

    2016-02-01

    In this work, in order to enhance the recovery performance of organic thin film transistors (OTFTs) ammonia (NH3) sensors, poly (3-hexylthiophene) (P3HT) and molybdenum disulfide (MoS2) were combined as sensitive materials. Different sensitive film structures as active layers of OTFTs, i.e., P3HT-MoS2 composite film, P3HT/MoS2 bilayer film and MoS2/P3HT bilayer film were fabricated by spray technology. OTFT gas sensors based on P3HT-MoS2 composite film showed a shorter recovery time than others when the ammonia concentration changed from 4 to 20 ppm. Specifically, x-ray diffraction (XRD), Raman and UV-visible absorption were employed to explore the interface properties between P3HT and single-layer MoS2. Through the complementary characterization, a mechanism based on charge transfer is proposed to explain the physical originality of these OTFT gas sensors: closer interlayer d-spacing and better π-π stacking of the P3HT chains in composite film have ensured a short recovery time of OTFT gas sensors. Moreover, sensing mechanisms of OTFTs were further studied by comparing the device performance in the presence of nitrogen or dry air as a carrier gas. This work not only strengthens the fundamental understanding of the sensing mechanism, but provides a promising approach to optimizing the OTFT gas sensors.

  2. Tunable Fabrication of Molybdenum Disulfide Quantum Dots for Intracellular MicroRNA Detection and Multiphoton Bioimaging.

    Science.gov (United States)

    Dai, Wenhao; Dong, Haifeng; Fugetsu, Bunshi; Cao, Yu; Lu, Huiting; Ma, Xinlei; Zhang, Xueji

    2015-09-01

    Molybdenum disulfide (MoS2 ) quantum dots (QDs) (size MoS2 QDs has not been investigated in great detail. Here, a facile and efficient approach for synthesis of controllable-size MoS2 QDs with excellent photoluminescence (PL) by using a sulfuric acid-assisted ultrasonic route is developed for this investigation. Various MoS2 structures including monolayer MoS2 flake, nanoporous MoS2 , and MoS2 QDs can be yielded by simply controlling the ultrasonic durations. Comprehensive microscopic and spectroscopic tools demonstrate that the MoS2 QDs have uniform lateral size and possess excellent excitation-independent blue PL. The as-generated MoS2 QDs show high quantum yield of 9.65%, long fluorescence lifetime of 4.66 ns, and good fluorescent stability over broad pH values from 4 to 10. Given the good intrinsic optical properties and large surface area combined with excellent physiological stability and biocompatibility, a MoS2 QDs-based intracellular microRNA imaging analysis system is successfully constructed. Importantly, the MoS2 QDs show good performance as multiphoton bioimaging labeling. The proposed synthesis strategy paves a new way for facile and efficient preparing MoS2 QDs with tunable-size for biomedical imaging and optoelectronic devices application. PMID:26033986

  3. A conserved cysteine residue is involved in disulfide bond formation between plant plasma membrane aquaporin monomers.

    Science.gov (United States)

    Bienert, Gerd P; Cavez, Damien; Besserer, Arnaud; Berny, Marie C; Gilis, Dimitri; Rooman, Marianne; Chaumont, François

    2012-07-01

    AQPs (aquaporins) are conserved in all kingdoms of life and facilitate the rapid diffusion of water and/or other small solutes across cell membranes. Among the different plant AQPs, PIPs (plasma membrane intrinsic proteins), which fall into two phylogenetic groups, PIP1 and PIP2, play key roles in plant water transport processes. PIPs form tetramers in which each monomer acts as a functional channel. The intermolecular interactions that stabilize PIP oligomer complexes and are responsible for the resistance of PIP dimers to denaturating conditions are not well characterized. In the present study, we identified a highly conserved cysteine residue in loop A of PIP1 and PIP2 proteins and demonstrated by mutagenesis that it is involved in the formation of a disulfide bond between two monomers. Although this cysteine seems not to be involved in regulation of trafficking to the plasma membrane, activity, substrate selectivity or oxidative gating of ZmPIP1s (Zm is Zea mays), ZmPIP2s and hetero-oligomers, it increases oligomer stability under denaturating conditions. In addition, when PIP1 and PIP2 are co-expressed, the loop A cysteine of ZmPIP1;2, but not that of ZmPIP2;5, is involved in the mercury sensitivity of the channels.

  4. Environmental Effects on Hysteresis of Transfer Characteristics in Molybdenum Disulfide Field-Effect Transistors

    Science.gov (United States)

    Shimazu, Yoshihiro; Tashiro, Mitsuki; Sonobe, Satoshi; Takahashi, Masaki

    2016-07-01

    Molybdenum disulfide (MoS2) has recently received much attention for nanoscale electronic and photonic applications. To explore the intrinsic properties and enhance the performance of MoS2-based field-effect transistors, thorough understanding of extrinsic effects such as environmental gas and contact resistance of the electrodes is required. Here, we report the effects of environmental gases on the transport properties of back-gated multilayered MoS2 field-effect transistors. Comparisons between different gases (oxygen, nitrogen, and air and nitrogen with varying relative humidities) revealed that water molecules acting as charge-trapping centers are the main cause of hysteresis in the transfer characteristics. While the hysteresis persisted even after pumping out the environmental gas for longer than 10 h at room temperature, it disappeared when the device was cooled to 240 K, suggesting a considerable increase in the time constant of the charge trapping/detrapping at these modestly low temperatures. The suppression of the hysteresis or instability in the easily attainable temperature range without surface passivation is highly advantageous for the device application of this system. The humidity dependence of the threshold voltages in the transfer curves indicates that the water molecules dominantly act as hole-trapping centers. A strong dependence of the on-state current on oxygen pressure was also observed.

  5. Mutations in the RAM network confer resistance to the thiol oxidant 4,4'-dipyridyl disulfide

    DEFF Research Database (Denmark)

    López-Mirabal, H Reynaldo; Winther, Jakob R; Thorsen, Michael;

    2008-01-01

    -specific oxidant dipyridyl disulfide (DPS) yielded tao3-516, which is impaired in the function of the RAM signaling network protein Tao3/Pag1p. We suggest that the DPS-resistance of the tao3-516 mutant might be due to deficient cell-cycle-regulated production of the chitinase Cts1p, which functions in post......-mitotic cell separation and depends on Tao3p and the RAM network for regulated expression. Consistent with this, deletion of other RAM genes or CTS1 also resulted in increased resistance to DPS. Exposure to DPS caused extensive depolarization of the actin cytoskeleton. We found that tao3-516 is resistant...... to latrunculin, a specific inhibitor of actin polymerization, and that ram, Deltaace2, and Deltacts1 mutants are resistant to benomyl, a microtubule-destabilizing drug. Since septum build-up depends on the organization of cytoskeletal proteins, the resistance to cytoskeletal stress of Cts1p-deficient mutants...

  6. Prophylaxis of Diallyl Disulfide on Skin Carcinogenic Model via p21-dependent Nrf2 stabilization

    Science.gov (United States)

    Shan, Yunlong; Wei, Zhonghong; Tao, Li; Wang, Siliang; Zhang, Feng; Shen, Cunsi; Wu, Hongyan; Liu, Zhaoguo; Zhu, Pingting; Wang, Aiyun; Chen, Wenxing; Lu, Yin

    2016-01-01

    Cancer prevention through intake of biologically active natural products appears to be an accessible way to reduce the risk of cancer. Diallyl disulfide (DADS), a major garlic derivative, has exhibited potential role in cancer therapy. The study is aimed to evaluate the prophylactic effect of DADS in chemically induced mouse skin carcinogenesis and investigate the molecular targets mediated by DADS. Two-stage chemically induced carcinogenesis model by cutaneous application of DMBA and subsequent TPA was established to study the prophylactic effect of DADS. As a result, we observed that DADS dose-dependently attenuated skin tumor incidence and multiplicity in the model mice, which was related to the up-regulation of a bunch of antioxidant enzymes activities and the nuclear accumulation of Nrf2. Furthermore, we developed skin carcinogenesis in Nrf2 knockout mice which could reverse the activity of DADS. Finally, we uncovered the underlying mechanism that DADS promoted the endogenous interaction between p21 and Nrf2, which was critical for impairing the Keap1-mediated degradation of Nrf2. Based on the results, we concluded that DADS was a promising cancer chemoprevention agent and suggested a garlic-rich diet might be beneficial to reduce the cancer risk in our daily life. PMID:27759091

  7. Hybrid quantum dot-tin disulfide field-effect transistors with improved photocurrent and spectral responsivity

    Science.gov (United States)

    Huang, Yuan; Zang, Huidong; Chen, Jia-Shiang; Sutter, Eli A.; Sutter, Peter W.; Nam, Chang-Yong; Cotlet, Mircea

    2016-03-01

    We report an improved photosensitivity in few-layer tin disulfide (SnS2) field-effect transistors (FETs) following doping with CdSe/ZnS core/shell quantum dots (QDs). The hybrid QD-SnS2 FET devices achieve more than 500% increase in the photocurrent response compared with the starting SnS2-only FET device and a spectral responsivity reaching over 650 A/W at 400 nm wavelength. The negligible electrical conductance in a control QD-only FET device suggests that the energy transfer between QDs and SnS2 is the main mechanism responsible for the sensitization effect, which is consistent with the strong spectral overlap between QD photoluminescence and SnS2 optical absorption as well as the large nominal donor-acceptor interspacing between QD core and SnS2. We also find enhanced charge carrier mobility in hybrid QD-SnS2 FETs which we attribute to a reduced contact Schottky barrier width due to an elevated background charge carrier density.

  8. Apoptosis induced by diallyl disulfide in human breast cancer cell line MCF.71

    Institute of Scientific and Technical Information of China (English)

    Xiao-yong LEI; Shu-qiong YAO; Xu-yu ZU; Ze-xiang HUANG; Li-juan LIU; Miao ZHONG; Bing-yang ZHU; Sheng- song TANG; Duan-fang LIAO

    2008-01-01

    Aim:To investigate the effect of diallyl disulfide (DADS),a component of garlic,on apoptosis in human mammary cancer cell line (MCF-7) and its mechanisms.Methods:Cytotoxicity was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays.Morphology of apoptotic cells was detected by acridine orange and ethidium bromide staining.Apoptotic cells stained with propidium iodide were examined using flow cytometry.Protein levels were detected by Western blot analysis.Results:DADS inhibited the proliferation of MCF-7 cells and induced the apoptotic ratio to increase rapidly.Cleavage of the caspase-3 and caspase-3 substrate poly(ADP-ribose) polymerase was observed in MCF-7 cells after 24 h of treatment with DADS.When the MCF-7 cells were signal-regulated kinase (ERK),a mitogen-activated protein kinase,was inhibited after 6 h; court N-terminal kinase (JNK),that is stress-activated protein kinase (SAPK),and p38 mitogen-aetivated protein kinase were activated after 6 h.Conclusion:These results suggest that DADS both inhibits the proliferation of MCF-7 cells and induces apoptosis of MCF-7 cells.The mechanisms may include the inhibition of ERK and the activation of the SAPK/JNK and p38 pathways.

  9. Contact research strategy for emerging molybdenum disulfide and other two-dimensional field-effect transistors

    Directory of Open Access Journals (Sweden)

    Yuchen Du

    2014-09-01

    Full Text Available Layered two-dimensional (2D semiconducting transition metal dichalcogenides (TMDs have been widely isolated, synthesized, and characterized recently. Numerous 2D materials are identified as the potential candidates as channel materials for future thin film technology due to their high mobility and the exhibiting bandgaps. While many TMD filed-effect transistors (FETs have been widely demonstrated along with a significant progress to clearly understand the device physics, large contact resistance at metal/semiconductor interface still remain a challenge. From 2D device research point of view, how to minimize the Schottky barrier effects on contacts thus reduce the contact resistance of metals on 2D materials is very critical for the further development of the field. Here, we present a review of contact research on molybdenum disulfide and other TMD FETs from the fundamental understanding of metal-semiconductor interfaces on 2D materials. A clear contact research strategy on 2D semiconducting materials is developed for future high-performance 2D FETs with aggressively scaled dimensions.

  10. Determination of density of states, conduction mechanisms and dielectric properties of nickel disulfide nanoparticles

    Science.gov (United States)

    Jamil, Arifa; Batool, S. S.; Sher, F.; Rafiq, M. A.

    2016-05-01

    Temperature and frequency dependent ac electrical measurements were used to explore density of states, conduction mechanisms and dielectric properties of nickel disulfide (NiS2) nanoparticles. The NiS2 nanoparticles were prepared by conventional one step solid state reaction method at 250 °C. X-ray diffraction (XRD) confirmed cubic phase of prepared nanoparticles. Scanning electron microscope (SEM) images revealed presence of irregular shaped nanoparticles as small as 50 nm. The ac electrical measurements were carried out from 300 K to 413 K. Two depressed semicircular arcs from 20 Hz to 2 MHz showed presence of bulk and grain boundary phases in NiS2 nanoparticles at all temperatures. Small polaron hopping conduction from 300 K to 393 K and correlated barrier hopping conduction mechanism at temperatures higher than 393 K was observed. High value of density of states (of the order of 1024 eV-1cm-3) was calculated from ac conductivity. At low frequencies high values (of the order of 104-107) of real part of dielectric constant (ɛ') were observed at different temperatures. These observations suggest that NiS2 nanoparticles may find applications in electronic devices.

  11. Utilizing self-assembled-monolayer-based gate dielectrics to fabricate molybdenum disulfide field-effect transistors

    Energy Technology Data Exchange (ETDEWEB)

    Kawanago, Takamasa, E-mail: kawanago.t.ab@m.titech.ac.jp; Oda, Shunri [Quantum Nanoelectronics Research Center (QNERC), Tokyo Institute of Technology, 2-12-1 Ookayama, Meguro-ku, Tokyo, 152-8550 (Japan)

    2016-01-25

    In this study, we apply self-assembled-monolayer (SAM)-based gate dielectrics to the fabrication of molybdenum disulfide (MoS{sub 2}) field-effect transistors. A simple fabrication process involving the selective formation of a SAM on metal oxides in conjunction with the dry transfer of MoS{sub 2} flakes was established. A subthreshold slope (SS) of 69 mV/dec and no hysteresis were demonstrated with the ultrathin SAM-based gate dielectrics accompanied by a low gate leakage current. The small SS and no hysteresis indicate the superior interfacial properties of the MoS{sub 2}/SAM structure. Cross-sectional transmission electron microscopy revealed a sharp and abrupt interface of the MoS{sub 2}/SAM structure. The SAM-based gate dielectrics are found to be applicable to the fabrication of low-voltage MoS{sub 2} field-effect transistors and can also be extended to various layered semiconductor materials. This study opens up intriguing possibilities of SAM-based gate dielectrics in functional electronic devices.

  12. Environmental Effects on Hysteresis of Transfer Characteristics in Molybdenum Disulfide Field-Effect Transistors.

    Science.gov (United States)

    Shimazu, Yoshihiro; Tashiro, Mitsuki; Sonobe, Satoshi; Takahashi, Masaki

    2016-07-20

    Molybdenum disulfide (MoS2) has recently received much attention for nanoscale electronic and photonic applications. To explore the intrinsic properties and enhance the performance of MoS2-based field-effect transistors, thorough understanding of extrinsic effects such as environmental gas and contact resistance of the electrodes is required. Here, we report the effects of environmental gases on the transport properties of back-gated multilayered MoS2 field-effect transistors. Comparisons between different gases (oxygen, nitrogen, and air and nitrogen with varying relative humidities) revealed that water molecules acting as charge-trapping centers are the main cause of hysteresis in the transfer characteristics. While the hysteresis persisted even after pumping out the environmental gas for longer than 10 h at room temperature, it disappeared when the device was cooled to 240 K, suggesting a considerable increase in the time constant of the charge trapping/detrapping at these modestly low temperatures. The suppression of the hysteresis or instability in the easily attainable temperature range without surface passivation is highly advantageous for the device application of this system. The humidity dependence of the threshold voltages in the transfer curves indicates that the water molecules dominantly act as hole-trapping centers. A strong dependence of the on-state current on oxygen pressure was also observed.

  13. Enhanced preclinical efficacy of tamoxifen developed as alginate-cysteine/disulfide bond reduced albumin nanoparticles.

    Science.gov (United States)

    Martínez, A; Muñiz, E; Iglesias, I; Teijón, J M; Blanco, M D

    2012-10-15

    Tamoxifen (TMX) is the most common clinical choice for the treatment of advanced or metastatic estrogen-dependent breast cancer. However, research on new challenging therapies is necessary due to its undesirable side effects and the limitation of the treatment only to the oral route. In this study, the antitumor activity of TMX-loaded nanoparticles based on different mixtures of alginate-cysteine and disulfide bond reduced bovine serum albumin was tested in vivo in MCF-7 nude mice xenograft model. These systems showed an enhancement of the TMX antitumor activity, since lower tumor evolutions and lower tumor growth rates were observed in mice treated with them. Moreover, histological and immunohistochemical studies revealed that treatments with TMX-loaded nanoparticles showed the most regressive and less proliferative tumor tissues. TMX biodistribution studies determined that TMX-loaded nanoparticles caused more accumulation of the drug into the tumor site with undetectable levels of TMX in plasma, reducing the possibility of delivering TMX to other not-targeted organs and, consequently, developing possible side effects. Thus, these TMX nanoparticulate systems are expected to provide a novel approach to the treatment of breast cancer in the future. PMID:22850290

  14. Environmental Effects on Hysteresis of Transfer Characteristics in Molybdenum Disulfide Field-Effect Transistors

    Science.gov (United States)

    Shimazu, Yoshihiro; Tashiro, Mitsuki; Sonobe, Satoshi; Takahashi, Masaki

    2016-01-01

    Molybdenum disulfide (MoS2) has recently received much attention for nanoscale electronic and photonic applications. To explore the intrinsic properties and enhance the performance of MoS2-based field-effect transistors, thorough understanding of extrinsic effects such as environmental gas and contact resistance of the electrodes is required. Here, we report the effects of environmental gases on the transport properties of back-gated multilayered MoS2 field-effect transistors. Comparisons between different gases (oxygen, nitrogen, and air and nitrogen with varying relative humidities) revealed that water molecules acting as charge-trapping centers are the main cause of hysteresis in the transfer characteristics. While the hysteresis persisted even after pumping out the environmental gas for longer than 10 h at room temperature, it disappeared when the device was cooled to 240 K, suggesting a considerable increase in the time constant of the charge trapping/detrapping at these modestly low temperatures. The suppression of the hysteresis or instability in the easily attainable temperature range without surface passivation is highly advantageous for the device application of this system. The humidity dependence of the threshold voltages in the transfer curves indicates that the water molecules dominantly act as hole-trapping centers. A strong dependence of the on-state current on oxygen pressure was also observed. PMID:27435309

  15. Platelet protein disulfide isomerase is required for thrombus formation but not for hemostasis in mice.

    Science.gov (United States)

    Kim, Kyungho; Hahm, Eunsil; Li, Jing; Holbrook, Lisa-Marie; Sasikumar, Parvathy; Stanley, Ronald G; Ushio-Fukai, Masuko; Gibbins, Jonathan M; Cho, Jaehyung

    2013-08-01

    Protein disulfide isomerase (PDI) derived from intravascular cells is required for thrombus formation. However, it remains unclear whether platelet PDI contributes to the process. Using platelet-specific PDI-deficient mice, we demonstrate that PDI-null platelets have defects in aggregation and adenosine triphosphate secretion induced by thrombin, collagen, and adenosine diphosphate. Such defects were rescued by wild-type but not mutant PDI, indicating that the isomerase activity of platelet surface PDI is critical for the regulatory effect. PDI-deficient platelets expressed increased levels of intracellular ER protein 57 (ERp57) and ERp72. Platelet PDI regulated αIIbβ3 integrin activation but not P-selectin exposure, Ca(2+) mobilization, β3-talin1 interaction, or platelet spreading on immobilized fibrinogen. Inhibition of ERp57 further diminished αIIbβ3 integrin activation and aggregation of activated PDI-deficient platelets, suggesting distinct roles of PDI and ERp57 in platelet functions. We found that platelet PDI is important for thrombus formation on collagen-coated surfaces under shear. Intravital microscopy demonstrates that platelet PDI is important for platelet accumulation but not initial adhesion and fibrin generation following laser-induced arteriolar injury. Tail bleeding time in platelet-specific PDI-deficient mice were not significantly increased. Our results provide important evidence that platelet PDI is essential for thrombus formation but not for hemostasis in mice. PMID:23788140

  16. Layer-modulated synthesis of uniform tungsten disulfide nanosheet using gas-phase precursors.

    Science.gov (United States)

    Park, Jusang; Lee, Wonseon; Choi, Taejin; Hwang, Sung-Hwan; Myoung, Jae Min; Jung, Jae-Hoon; Kim, Soo-Hyun; Kim, Hyungjun

    2015-01-28

    The synthesis of layered transition-metal-disulfide (MS2, M = Mo, W) nanosheets with layer controllability and large-area uniformity is an essential requirement for their application in electronic and optical devices. In this report, we describe a synthesis process of WS2 nanosheets with layer controllability and high uniformity using chemical vapor deposition (CVD) and WCl6 and H2S as gas-phase precursors. Through this process, we can systematically modulate the thickness of WS2 nanosheets by controlling the duration of the reaction between WCl6 and H2S. The CVD-grown WS2 nanosheets exhibit good stoichiometry as well as dependencies of a clear Raman shift and bandgap on the number of layers. These properties are confirmed by X-ray photoemission spectroscopy, Raman spectroscopy, and photoluminescence measurements. The number of layers of WS2 nanosheets is confirmed by atomic force microscopy. Finally, we demonstrate the fabrication and performance of a photodetector based on a hybrid structure consisting of graphene and a WS2 nanosheet.

  17. Layer-modulated synthesis of uniform tungsten disulfide nanosheet using gas-phase precursors.

    Science.gov (United States)

    Park, Jusang; Lee, Wonseon; Choi, Taejin; Hwang, Sung-Hwan; Myoung, Jae Min; Jung, Jae-Hoon; Kim, Soo-Hyun; Kim, Hyungjun

    2015-01-28

    The synthesis of layered transition-metal-disulfide (MS2, M = Mo, W) nanosheets with layer controllability and large-area uniformity is an essential requirement for their application in electronic and optical devices. In this report, we describe a synthesis process of WS2 nanosheets with layer controllability and high uniformity using chemical vapor deposition (CVD) and WCl6 and H2S as gas-phase precursors. Through this process, we can systematically modulate the thickness of WS2 nanosheets by controlling the duration of the reaction between WCl6 and H2S. The CVD-grown WS2 nanosheets exhibit good stoichiometry as well as dependencies of a clear Raman shift and bandgap on the number of layers. These properties are confirmed by X-ray photoemission spectroscopy, Raman spectroscopy, and photoluminescence measurements. The number of layers of WS2 nanosheets is confirmed by atomic force microscopy. Finally, we demonstrate the fabrication and performance of a photodetector based on a hybrid structure consisting of graphene and a WS2 nanosheet. PMID:25361429

  18. Cathode based on molybdenum disulfide nanoflakes for lithium-oxygen batteries.

    Energy Technology Data Exchange (ETDEWEB)

    Asadi, Mohammad; Kumar, Bijandra; Liu, Cong; Phillips, Patrick; Yasaei, Poya; Behranginia, Amirhossein; Zapol, Peter; Klie, Robert F.; Curtiss, Larry A.; Salehi-Khojin, Amin

    2016-02-01

    Lithium-oxygen (Li-O-2) batteries have been recognized as an emerging technology for energy storage systems owing to their high theoretical specific energy. One challenge is to find an electrolyte/cathode system that is efficient, stable, and cost-effective. We present such a system based on molybdenum disulfide (MoS2) nanoflakes combined with an ionic liquid (IL) that work together as an effective cocatalyst for discharge and charge in a Li-O-2 battery. Cyclic voltammetry results show superior catalytic performance for this cocatalyst for both oxygen reduction and evolution reactions compared to Au and Pt catalysts. It also performs remarkably well in the Li-O-2 battery system with 85% round-trip efficiency and reversibility up to 50 cycles. Density functional calculations provide a mechanistic understanding of the MoS2 nanoflakes/IL system. cocatalyst reported in this work could open the way for exploiting the unique properties of ionic liquids in Li-air batteries in combination with nanostructured MoS2 as a cathode material.

  19. Enhancement of protein secretion in Pichia pastoris by overexpression of protein disulfide isomerase.

    Science.gov (United States)

    Inan, Mehmet; Aryasomayajula, Dinesh; Sinha, Jayanta; Meagher, Michael M

    2006-03-01

    A potential vaccine candidate, Necator americanus secretory protein (Na-ASP1), against hookworm infections, has been expressed in Pichia pastoris. Na-ASP1, a 45 kDa protein containing 20 cysteines, was directed outside the cell by fusing the protein to the preprosequence of the alpha-mating factor of Saccharomyces cerevisiae. Most of the protein produced by single copy clones was secreted outside the cell. However, increasing gene copy number of Na-ASP1 protein in P. pastoris saturated secretory capacity and therefore, decreased the amount of secreted protein in clones harboring multiple copies of Na-ASP1 gene. Overexpression of the endoplasmic reticulum (ER) resident, homologous chaperone protein, protein disulfide isomerase (PDI) was able to increase the secretion of (Na-ASP1) protein in high copy clones. The effect of PDI levels on secretion of Na-ASP1 protein was examined in clones with varying copy number of PDI gene. Increase in secreted Na-ASP1 secretion is correlated well with the PDI copy number. Increasing levels of PDI also increased overall Na-ASP1 protein production in all the clones. Nevertheless, there was still accumulation of intracellular Na-ASP1 protein in P. pastoris clones over-expressing Na-ASP1 and PDI proteins. PMID:16255058

  20. Few-layer molybdenum disulfide transistors and circuits for high-speed flexible electronics.

    Science.gov (United States)

    Cheng, Rui; Jiang, Shan; Chen, Yu; Liu, Yuan; Weiss, Nathan; Cheng, Hung-Chieh; Wu, Hao; Huang, Yu; Duan, Xiangfeng

    2014-10-08

    Two-dimensional layered materials, such as molybdenum disulfide, are emerging as an exciting material system for future electronics due to their unique electronic properties and atomically thin geometry. Here we report a systematic investigation of MoS2 transistors with optimized contact and device geometry, to achieve self-aligned devices with performance including an intrinsic gain over 30, an intrinsic cut-off frequency fT up to 42 GHz and a maximum oscillation frequency fMAX up to 50 GHz, exceeding the reported values for MoS2 transistors to date (fT~0.9 GHz, fMAX~1 GHz). Our results show that logic inverters or radio frequency amplifiers can be formed by integrating multiple MoS2 transistors on quartz or flexible substrates with voltage gain in the gigahertz regime. This study demonstrates the potential of two-dimensional layered semiconductors for high-speed flexible electronics.

  1. Chemical vapor deposition based tungsten disulfide (WS2) thin film transistor

    KAUST Repository

    Hussain, Aftab M.

    2013-04-01

    Tungsten disulfide (WS2) is a layered transition metal dichalcogenide with a reported band gap of 1.8 eV in bulk and 1.32-1.4 eV in its thin film form. 2D atomic layers of metal dichalcogenides have shown changes in conductivity with applied electric field. This makes them an interesting option for channel material in field effect transistors (FETs). Therefore, we show a highly manufacturable chemical vapor deposition (CVD) based simple process to grow WS2 directly on silicon oxide in a furnace and then its transistor action with back gated device with room temperature field effect mobility of 0.1003 cm2/V-s using the Schottky barrier contact model. We also show the semiconducting behavior of this WS2 thin film which is more promising than thermally unstable organic materials for thin film transistor application. Our direct growth method on silicon oxide also holds interesting opportunities for macro-electronics applications. © 2013 IEEE.

  2. Tribological properties of adaptive phosphate composite coatings with addition of silver and molybdenum disulfide

    Science.gov (United States)

    Liu, Cancan; Chen, Lei; Zhou, Jiansong; Zhou, Huidi; Chen, Jianmin

    2014-05-01

    Adaptive phosphate composite coatings with addition of solid lubricants of molybdenum disulfide (MoS2) and silver (Ag) using aluminum chromium phosphate as the binder were fabricated on high-temperature steel. The tribological properties of phosphate composite coatings were evaluated from room temperature (RT) to 700 °C. The phase composition and microstructure were investigated according to the characterization by power X-ray diffraction (XRD), Raman spectroscopy and scanning electron microscopy (SEM). The results show that the composite coating with the Ag/MoS2 mass ratio of 2:1 exhibits the stable and low friction coefficients from RT to 700 °C and relative low wear rates at all testing temperatures. The tribo-chemical reaction between Ag and MoS2 occurred in the rubbing process to form silver molybdates compounds lubricating film. The temperature-adaptive tribological properties were attributed to the formation of lubricating films composed of lubricants silver, MoS2 and silver molybdates phases on the worn surfaces of the composites coatings in a wide-temperature range.

  3. Mapping Soluble Guanylyl Cyclase and Protein Disulfide Isomerase Regions of Interaction.

    Directory of Open Access Journals (Sweden)

    Erin J Heckler

    Full Text Available Soluble guanylyl cyclase (sGC is a heterodimeric nitric oxide (NO receptor that produces cyclic GMP. This signaling mechanism is a key component in the cardiovascular system. NO binds to heme in the β subunit and stimulates the catalytic conversion of GTP to cGMP several hundred fold. Several endogenous factors have been identified that modulate sGC function in vitro and in vivo. In previous work, we determined that protein disulfide isomerase (PDI interacts with sGC in a redox-dependent manner in vitro and that PDI inhibited NO-stimulated activity in cells. To our knowledge, this was the first report of a physical interaction between sGC and a thiol-redox protein. To characterize this interaction between sGC and PDI, we first identified peptide linkages between sGC and PDI, using a lysine cross-linking reagent and recently developed mass spectrometry analysis. Together with Flag-immunoprecipitation using sGC domain deletions, wild-type (WT and mutated PDI, regions of sGC involved in this interaction were identified. The observed data were further explored with computational modeling to gain insight into the interaction mechanism between sGC and oxidized PDI. Our results indicate that PDI interacts preferentially with the catalytic domain of sGC, thus providing a mechanism for PDI inhibition of sGC. A model in which PDI interacts with either the α or the β catalytic domain is proposed.

  4. Purification, crystallization and preliminary crystallographic investigation of FrnE, a disulfide oxidoreductase from Deinococcus radiodurans.

    Science.gov (United States)

    Panicker, Lata; Misra, Hari Sharan; Bihani, Subhash Chandra

    2014-11-01

    In prokaryotes, Dsb proteins catalyze the formation of native disulfide bonds through an oxidative folding pathway and are part of the cell machinery that protects proteins from oxidative stress. Deinococcus radiodurans is an extremophile which shows unparalleled resistance to ionizing radiation and oxidative stress. It has a strong mechanism to protect its proteome from oxidative damage. The genome of Deinococcus shows the presence of FrnE, a Dsb protein homologue that potentially provides the bacterium with oxidative stress tolerance. Here, crystallization and preliminary X-ray crystallographic analysis of FrnE from D. radiodurans are reported. Diffraction-quality single crystals were obtained using the hanging-drop vapour-diffusion method with reservoir solution consisting of 100 mM sodium acetate pH 5.0, 10% PEG 8000, 15-20% glycerol. Diffraction data were collected on an Agilent SuperNova system using a microfocus sealed-tube X-ray source. The crystal diffracted to 1.8 Å resolution at 100 K. The space group of the crystal was found to be P2₁22₁, with unit-cell parameters a=47.91, b=62.94, c=86.75 Å, α=β=γ=90°. Based on Matthews coefficient analysis, one monomer per asymmetric unit is present in the crystal, with a solvent content of approximately 45%.

  5. Utilizing self-assembled-monolayer-based gate dielectrics to fabricate molybdenum disulfide field-effect transistors

    International Nuclear Information System (INIS)

    In this study, we apply self-assembled-monolayer (SAM)-based gate dielectrics to the fabrication of molybdenum disulfide (MoS2) field-effect transistors. A simple fabrication process involving the selective formation of a SAM on metal oxides in conjunction with the dry transfer of MoS2 flakes was established. A subthreshold slope (SS) of 69 mV/dec and no hysteresis were demonstrated with the ultrathin SAM-based gate dielectrics accompanied by a low gate leakage current. The small SS and no hysteresis indicate the superior interfacial properties of the MoS2/SAM structure. Cross-sectional transmission electron microscopy revealed a sharp and abrupt interface of the MoS2/SAM structure. The SAM-based gate dielectrics are found to be applicable to the fabrication of low-voltage MoS2 field-effect transistors and can also be extended to various layered semiconductor materials. This study opens up intriguing possibilities of SAM-based gate dielectrics in functional electronic devices

  6. Environmental Effects on Hysteresis of Transfer Characteristics in Molybdenum Disulfide Field-Effect Transistors.

    Science.gov (United States)

    Shimazu, Yoshihiro; Tashiro, Mitsuki; Sonobe, Satoshi; Takahashi, Masaki

    2016-01-01

    Molybdenum disulfide (MoS2) has recently received much attention for nanoscale electronic and photonic applications. To explore the intrinsic properties and enhance the performance of MoS2-based field-effect transistors, thorough understanding of extrinsic effects such as environmental gas and contact resistance of the electrodes is required. Here, we report the effects of environmental gases on the transport properties of back-gated multilayered MoS2 field-effect transistors. Comparisons between different gases (oxygen, nitrogen, and air and nitrogen with varying relative humidities) revealed that water molecules acting as charge-trapping centers are the main cause of hysteresis in the transfer characteristics. While the hysteresis persisted even after pumping out the environmental gas for longer than 10 h at room temperature, it disappeared when the device was cooled to 240 K, suggesting a considerable increase in the time constant of the charge trapping/detrapping at these modestly low temperatures. The suppression of the hysteresis or instability in the easily attainable temperature range without surface passivation is highly advantageous for the device application of this system. The humidity dependence of the threshold voltages in the transfer curves indicates that the water molecules dominantly act as hole-trapping centers. A strong dependence of the on-state current on oxygen pressure was also observed. PMID:27435309

  7. Sodium and Lithium Storage Properties of Spray-Dried Molybdenum Disulfide-Graphene Hierarchical Microspheres

    Science.gov (United States)

    Kalluri, Sujith; Seng, Kuok Hau; Guo, Zaiping; Du, Aijun; Konstantinov, Konstantin; Liu, Hua Kun; Dou, Shi Xue

    2015-07-01

    Developing nano/micro-structures which can effectively upgrade the intriguing properties of electrode materials for energy storage devices is always a key research topic. Ultrathin nanosheets were proved to be one of the potential nanostructures due to their high specific surface area, good active contact areas and porous channels. Herein, we report a unique hierarchical micro-spherical morphology of well-stacked and completely miscible molybdenum disulfide (MoS2) nanosheets and graphene sheets, were successfully synthesized via a simple and industrial scale spray-drying technique to take the advantages of both MoS2 and graphene in terms of their high practical capacity values and high electronic conductivity, respectively. Computational studies were performed to understand the interfacial behaviour of MoS2 and graphene, which proves high stability of the composite with high interfacial binding energy (-2.02 eV) among them. Further, the lithium and sodium storage properties have been tested and reveal excellent cyclic stability over 250 and 500 cycles, respectively, with the highest initial capacity values of 1300 mAh g-1 and 640 mAh g-1 at 0.1 A g-1.

  8. Preparation of titanium dioxide/tungsten disulfide composite photocatalysts with enhanced photocatalytic activity under visible light

    Energy Technology Data Exchange (ETDEWEB)

    Zheng, Lili; Zhang, Weiping; Xiao, Xinyan [South China University of Technology, Guangzhou (China)

    2016-01-15

    Titanium dioxide/tungsten disulfide (TiO{sub 2}/WS{sub 2}) composite photocatalysts were fabricated via a one-step hydrothermal synthesis process, using TiCl{sub 4} as titanium source and bulk WS{sub 2} as sensitizer. The morphology, structure, specific surface area and optical absorption properties of the composite photocatalysts were characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FTIR), X-ray powder diffraction (XRD), specific surface area analyzer and ultraviolet-visible diffuse reflection spectrum (UV-vis DRS), respectively. The photocatalytic activity of as-prepared photocatalysts was evaluated by the degradation of methyl orange (MO) under illumination of 500W Xenon lamp. The results indicated that TiO2/WS2 composite photocatalysts possessed excellent photocatalytic activity, and -95% of the degradation rate for MO was reached when molar ratio of WS{sub 2} to TiO{sub 2} was 0.004 and the irradiation time was 60 min. Moreover, the carrier trapping experiment and fluorescence spectra showed that •O{sup -}{sub 2} was the key component in the photocatalytic degradation process and O{sub 2} was reduced to be •O{sup -}{sub 2} by the electrons from the conduction band of TiO{sub 2} and WS{sub 2} for the degradation of MO.

  9. 40 CFR 63.500 - Back-end process provisions-carbon disulfide limitations for styrene butadiene rubber by emulsion...

    Science.gov (United States)

    2010-07-01

    .... There shall be a standard operating procedure representing the production of every grade of styrene... or 1A of 40 CFR part 60, appendix A, as required, shall be used for selection of the sampling sites. (ii) The gas volumetric flow rate shall be determined using Method 2, 2A, 2C, or 2D of 40 CFR part...

  10. The α-proteobacteria Wolbachia pipientis protein disulfide machinery has a regulatory mechanism absent in γ-proteobacteria.

    Directory of Open Access Journals (Sweden)

    Patricia M Walden

    Full Text Available The α-proteobacterium Wolbachia pipientis infects more than 65% of insect species worldwide and manipulates the host reproductive machinery to enable its own survival. It can live in mutualistic relationships with hosts that cause human disease, including mosquitoes that carry the Dengue virus. Like many other bacteria, Wolbachia contains disulfide bond forming (Dsb proteins that introduce disulfide bonds into secreted effector proteins. The genome of the Wolbachia strain wMel encodes two DsbA-like proteins sharing just 21% sequence identity to each other, α-DsbA1 and α-DsbA2, and an integral membrane protein, α-DsbB. α-DsbA1 and α-DsbA2 both have a Cys-X-X-Cys active site that, by analogy with Escherichia coli DsbA, would need to be oxidized to the disulfide form to serve as a disulfide bond donor toward substrate proteins. Here we show that the integral membrane protein α-DsbB oxidizes α-DsbA1, but not α-DsbA2. The interaction between α-DsbA1 and α-DsbB is very specific, involving four essential cysteines located in the two periplasmic loops of α-DsbB. In the electron flow cascade, oxidation of α-DsbA1 by α-DsbB is initiated by an oxidizing quinone cofactor that interacts with the cysteine pair in the first periplasmic loop. Oxidizing power is transferred to the second cysteine pair, which directly interacts with α-DsbA1. This reaction is inhibited by a non-catalytic disulfide present in α-DsbA1, conserved in other α-proteobacterial DsbAs but not in γ-proteobacterial DsbAs. This is the first characterization of the integral membrane protein α-DsbB from Wolbachia and reveals that the non-catalytic cysteines of α-DsbA1 regulate the redox relay system in cooperation with α-DsbB.

  11. An efficient algorithmic approach for mass spectrometry-based disulfide connectivity determination using multi-ion analysis

    Directory of Open Access Journals (Sweden)

    Yen Ten-Yang

    2011-02-01

    Full Text Available Abstract Background Determining the disulfide (S-S bond pattern in a protein is often crucial for understanding its structure and function. In recent research, mass spectrometry (MS based analysis has been applied to this problem following protein digestion under both partial reduction and non-reduction conditions. However, this paradigm still awaits solutions to certain algorithmic problems fundamental amongst which is the efficient matching of an exponentially growing set of putative S-S bonded structural alternatives to the large amounts of experimental spectrometric data. Current methods circumvent this challenge primarily through simplifications, such as by assuming only the occurrence of certain ion-types (b-ions and y-ions that predominate in the more popular dissociation methods, such as collision-induced dissociation (CID. Unfortunately, this can adversely impact the quality of results. Method We present an algorithmic approach to this problem that can, with high computational efficiency, analyze multiple ions types (a, b, bo, b*, c, x, y, yo, y*, and z and deal with complex bonding topologies, such as inter/intra bonding involving more than two peptides. The proposed approach combines an approximation algorithm-based search formulation with data driven parameter estimation. This formulation considers only those regions of the search space where the correct solution resides with a high likelihood. Putative disulfide bonds thus obtained are finally combined in a globally consistent pattern to yield the overall disulfide bonding topology of the molecule. Additionally, each bond is associated with a confidence score, which aids in interpretation and assimilation of the results. Results The method was tested on nine different eukaryotic Glycosyltransferases possessing disulfide bonding topologies of varying complexity. Its performance was found to be characterized by high efficiency (in terms of time and the fraction of search space

  12. Graphene-Molybdenum Disulfide-Graphene Tunneling Junctions with Large-Area Synthesized Materials.

    Science.gov (United States)

    Joiner, Corey A; Campbell, Philip M; Tarasov, Alexey A; Beatty, Brian R; Perini, Chris J; Tsai, Meng-Yen; Ready, William J; Vogel, Eric M

    2016-04-01

    Tunneling devices based on vertical heterostructures of graphene and other 2D materials can overcome the low on-off ratios typically observed in planar graphene field-effect transistors. This study addresses the impact of processing conditions on two-dimensional materials in a fully integrated heterostructure device fabrication process. In this paper, graphene-molybdenum disulfide-graphene tunneling heterostructures were fabricated using only large-area synthesized materials, unlike previous studies that used small exfoliated flakes. The MoS2 tunneling barrier is either synthesized on a sacrificial substrate and transferred to the bottom-layer graphene or synthesized directly on CVD graphene. The presence of graphene was shown to have no impact on the quality of the grown MoS2. The thickness uniformity of MoS2 grown on graphene and SiO2 was found to be 1.8 ± 0.22 nm. XPS and Raman spectroscopy are used to show how the MoS2 synthesis process introduces defects into the graphene structure by incorporating sulfur into the graphene. The incorporation of sulfur was shown to be greatly reduced in the absence of molybdenum suggesting molybdenum acts as a catalyst for sulfur incorporation. Tunneling simulations based on the Bardeen transfer Hamiltonian were performed and compared to the experimental tunneling results. The simulations show the use of MoS2 as a tunneling barrier suppresses contributions to the tunneling current from the conduction band. This is a result of the observed reduction of electron conduction within the graphene sheets. PMID:26987383

  13. Protein disulfide isomerase as a novel target for cyclopentenone prostaglandins: implications for hypoxic ischemic injury

    Science.gov (United States)

    Liu, Hao; Chen, Jie; Li, Wenjin; Rose, Marie E.; Shinde, Sunita N.; Balasubramani, Manimalha; Uechi, Guy T.; Mutus, Bülent; Graham, Steven H.; Hickey, Robert W.

    2016-01-01

    Cyclooxygenase-2 (COX-2) is an important contributor to ischemic brain injury. Identification of the downstream mediators of COX-2 toxicity may allow the development of targeted therapies. Of particular interest is the cyclopentenone family of prostaglandin metabolites. Cyclopentenone prostaglandins (CyPGs) are highly reactive molecules that form covalent bonds with cellular thiols. Protein disulfide isomerase (PDI) is an important molecule for the restoration of denatured proteins following ischemia. Because PDI has several thiols, including thiols within the active thioredoxin-like domain, we hypothesized that PDI is a target of CyPGs and that CyPG binding of PDI is detrimental. CyPG–PDI binding was detected in vitro via immunoprecipitation and MS. CyPG–PDI binding decreased PDI enzymatic activity in recombinant PDI treated with CyPG, and PDI immunoprecipitated from neuronal culture treated with CyPG or anoxia. Toxic effects of binding were demonstrated in experiments showing that: (a) pharmacologic inhibition of PDI increased cell death in anoxic neurons, (b) PDI overexpression protected neurons exposed to anoxia and SH-SY5Y cells exposed to CyPG, and (c) PDI overexpression in SH-SY5Y cells attenuated ubiquitination of proteins and decreased activation of pro-apoptotic caspases. In conclusion, CyPG production and subsequent binding of PDI is a novel and potentially important mechanism of ischemic brain injury. We show that CyPGs bind to PDI, cyclopentenones inhibit PDI activity, and CyPG–PDI binding is associated with increased neuronal susceptibility to anoxia. Additional studies are necessary to determine the relative role of CyPG-dependent inhibition of PDI activity in ischemia and other neurodegenerative disorders. PMID:25754985

  14. Effect of diallyl disulfide on acute gastric mucosal damage induced by alcohol in rats.

    Science.gov (United States)

    Lee, I-C; Baek, H-S; Kim, S-H; Moon, C; Park, S-H; Kim, S-H; Shin, I-S; Park, S-C; Kim, J-C

    2015-03-01

    This study investigated the gastroprotective effects of diallyl disulfide (DADS), a secondary organosulfur compound derived from garlic (Allium sativum L.) on experimental model of ethanol (EtOH)-induced gastric ulcer in rats. The antiulcerogenic activity of DADS was evaluated by gross/histopathological inspection, pro-inflammatory cytokines, and lipid peroxidation with antioxidant enzyme activities in the stomach. DADS (100 mg/kg) was administered by oral gavage 2 h prior to EtOH treatment (5 ml/kg). The animals were killed 1 h after receiving EtOH treatment. Pretreatment with DADS attenuated EtOH-induced gastric mucosal injury, as evidenced by decreased severity of hemorrhagic lesions and gastric ulcer index upon visual inspection. DADS also prevented histopathological alterations and gastric apoptotic changes caused by EtOH. An increase in tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase was observed in the gastric tissues of EtOH-treated rats that coincided with increased serum TNF-α and interleukin 6 levels. In contrast, DADS effectively suppressed production of pro-inflammatory mediators induced by EtOH. Furthermore, DADS prevented the formation of gastric malondialdehyde and the depletion of reduced glutathione content and restored antioxidant enzyme activities, such as catalase, glutathione peroxidase, and glutathione reductase in the gastric tissues of EtOH-treated rats. These results indicate that DADS prevents gastric mucosal damage induced by acute EtOH administration in rats and that the protective effects of DADS may be due to its potent antioxidant and anti-inflammatory activities. PMID:24972622

  15. Molybdenum disulfide nanoflower-chitosan-Au nanoparticles composites based electrochemical sensing platform for bisphenol A determination

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Ke-Jing, E-mail: kejinghuang@163.com [College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000 (China); State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082 (China); Liu, Yu-Jie; Liu, Yan-Ming; Wang, Ling-Ling [College of Chemistry and Chemical Engineering, Xinyang Normal University, Xinyang 464000 (China)

    2014-07-15

    Highlights: • This work constructs a novel electrochemical biosensor for bisphenol A detection. • Flower-like MoS{sub 2} are prepared by a simple hydrothermal procedure. • AuNPs are assembled on MoS{sub 2} nanoflowers modified electrode for signal amplification. • The developed sensor exhibits low detection limit and wide linear range. - Abstract: Two-dimensional transition metal dichalcogenide are attracting increasing attention in electrochemical sensing due to their unique electronic properties. In this work, flower-like molybdenum disulfide (MoS{sub 2}) was prepared by a simple hydrothermal method. The scanning electron microscopy and transmission electron microscopy images showed the MoS{sub 2} nanoflower had sizes with diameter of about 200 nm and was constructed with many irregular sheets as a petal-like structure with thickness of several nanometers. A novel electrochemical sensor was constructed for the determination of bisphenol A (BPA) based on MoS{sub 2} and chitosan-gold nanoparticles composites modified electrode. The sensor showed an efficient electrocatalytic role for the oxidation of BPA, and the oxidation overpotentials of BPA decreased significantly and the peak current increased greatly compared with bare GCE and other modified electrode. A good linear relationship between the oxidation peak current and BPA concentration was obtained in the range from 0.05 to 100 μM with a detection limit of 5.0 × 10{sup −9} M (S/N = 3). The developed sensor exhibited high sensitivity and long-term stability, and it was successfully applied for the determination of BPA in different samples. This work indicated MoS{sub 2} nanoflowers were promising in electrochemical sensing and catalytic applications.

  16. Molybdenum disulfide nanoflower-chitosan-Au nanoparticles composites based electrochemical sensing platform for bisphenol A determination

    International Nuclear Information System (INIS)

    Highlights: • This work constructs a novel electrochemical biosensor for bisphenol A detection. • Flower-like MoS2 are prepared by a simple hydrothermal procedure. • AuNPs are assembled on MoS2 nanoflowers modified electrode for signal amplification. • The developed sensor exhibits low detection limit and wide linear range. - Abstract: Two-dimensional transition metal dichalcogenide are attracting increasing attention in electrochemical sensing due to their unique electronic properties. In this work, flower-like molybdenum disulfide (MoS2) was prepared by a simple hydrothermal method. The scanning electron microscopy and transmission electron microscopy images showed the MoS2 nanoflower had sizes with diameter of about 200 nm and was constructed with many irregular sheets as a petal-like structure with thickness of several nanometers. A novel electrochemical sensor was constructed for the determination of bisphenol A (BPA) based on MoS2 and chitosan-gold nanoparticles composites modified electrode. The sensor showed an efficient electrocatalytic role for the oxidation of BPA, and the oxidation overpotentials of BPA decreased significantly and the peak current increased greatly compared with bare GCE and other modified electrode. A good linear relationship between the oxidation peak current and BPA concentration was obtained in the range from 0.05 to 100 μM with a detection limit of 5.0 × 10−9 M (S/N = 3). The developed sensor exhibited high sensitivity and long-term stability, and it was successfully applied for the determination of BPA in different samples. This work indicated MoS2 nanoflowers were promising in electrochemical sensing and catalytic applications

  17. Post-streptococcal auto-antibodies inhibit protein disulfide isomerase and are associated with insulin resistance.

    Directory of Open Access Journals (Sweden)

    Adi Aran

    Full Text Available Post-streptococcal autoimmunity affects millions worldwide, targeting multiple organs including the heart, brain, and kidneys. To explore the post-streptococcal autoimmunity spectrum, we used western blot analyses, to screen 310 sera from healthy subjects with (33% and without (67% markers of recent streptococcal infections [anti-Streptolysin O (ASLO or anti-DNAse B (ADB]. A 58 KDa protein, reacting strongly with post-streptococcal sera, was identified as Protein Disulfide Isomerase (PDI, an abundant protein with pleiotropic metabolic, immunologic, and thrombotic effects. Anti-PDI autoantibodies, purified from human sera, targeted similar epitopes in Streptolysin O (SLO, P51-61 and PDI (P328-338. The correlation between post-streptococcal status and anti-human PDI auto-immunity was further confirmed in a total of 2987 samples (13.6% in 530 ASLO positive versus 5.6% in 2457 ASLO negative samples, p<0.0001. Finally, anti-PDI auto-antibodies inhibited PDI-mediated insulin degradation in vitro (n = 90, p<0.001, and correlated with higher serum insulin (14.1 iu/ml vs. 12.2 iu/ml, n = 1215, p = 0.039 and insulin resistance (Homeostatic Model Assessment (HOMA 4.1 vs. 3.1, n = 1215, p = 0.004, in a population-based cohort. These results identify PDI as a major target of post-streptococcal autoimmunity, and establish a new link between infection, autoimmunity, and metabolic disturbances.

  18. Patagonfibrase modifies protein expression of tissue factor and protein disulfide isomerase in rat skin.

    Science.gov (United States)

    Peichoto, María Elisa; Santoro, Marcelo Larami

    2016-09-01

    Patagonfibrase is a hemorrhagic metalloproteinase isolated from the venom of the South American rear-fanged snake Philodryas patagoniensis, and is an important contributor to local lesions inflicted by this species. The tissue factor (TF)-factor VIIa complex, besides triggering the coagulation cascade, has been demonstrated to be involved in inflammatory events. Our aim was to determine whether patagonfibrase affects the expression of TF and protein disulfide isomerase (PDI), an enzyme that controls TF biological activity, at the site of patagonfibrase injection, and thus if they may play a role in hemostatic and inflammatory events induced by snake venoms. Patagonfibrase (60 μg/kg) was administered s.c. to rats, and after 3 h blood was collected to evaluate hemostasis parameters, and skin fragments close to the site of injection were taken to assess TF and PDI expression. Patagonfibrase did not alter blood cell counts, plasma fibrinogen levels, or levels of TF activity in plasma. However, by semiquantitative Western blotting, patagonfibrase increased TF expression by 2-fold, and decreased PDI expression by 3-fold in skin samples. In agreement, by immunohistochemical analyses, prominent TF expression was observed in the subcutaneous tissue. Thus, patagonfibrase affects the local expression of TF and PDI without inducing any systemic hemostatic disturbance, although that they may be involved in the local inflammatory events induced by hemorrhagic metalloproteinases. Once antivenom therapy is not totally effective to treat the local injury induced by snake venoms, modulation of the activity and expression of TF and/or PDI might become a strategy for treating snake envenomation. PMID:27390042

  19. Tribological properties of adaptive phosphate composite coatings with addition of silver and molybdenum disulfide

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Cancan [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou (China); University of Chinese Academy of Sciences, Beijing (China); Chen, Lei, E-mail: chenlei@lzb.ac.cn [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou (China); Zhou, Jiansong, E-mail: jszhou@licp.cas.cn [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou (China); Zhou, Huidi; Chen, Jianmin [State Key Laboratory of Solid Lubrication, Lanzhou Institute of Chemical Physics, Chinese Academy of Sciences, Lanzhou (China)

    2014-05-01

    Highlights: • A new kind of adaptive coatings was fabricated using relatively simple spraying techniques. • The tribological properties of Ag/MoS{sub 2} phosphate composite coatings were investigated at the temperature from 20 °C to 700 °C. • The composition and wear mechanisms of Ag/MoS{sub 2} phosphate composite coatings were also discussed. • The Ag/MoS{sub 2} phosphate composite coatings have self-repairing capability in the rubbing process at 700 °C. - Abstract: Adaptive phosphate composite coatings with addition of solid lubricants of molybdenum disulfide (MoS{sub 2}) and silver (Ag) using aluminum chromium phosphate as the binder were fabricated on high-temperature steel. The tribological properties of phosphate composite coatings were evaluated from room temperature (RT) to 700 °C. The phase composition and microstructure were investigated according to the characterization by power X-ray diffraction (XRD), Raman spectroscopy and scanning electron microscopy (SEM). The results show that the composite coating with the Ag/MoS{sub 2} mass ratio of 2:1 exhibits the stable and low friction coefficients from RT to 700 °C and relative low wear rates at all testing temperatures. The tribo-chemical reaction between Ag and MoS{sub 2} occurred in the rubbing process to form silver molybdates compounds lubricating film. The temperature-adaptive tribological properties were attributed to the formation of lubricating films composed of lubricants silver, MoS{sub 2} and silver molybdates phases on the worn surfaces of the composites coatings in a wide-temperature range.

  20. A cell nanoinjector based on carbon nanotubes

    Energy Technology Data Exchange (ETDEWEB)

    Chen, Xing; Kis, Andras; Zettl, Alex; Bertozzi, Carolyn R.

    2007-01-30

    Technologies for introducing molecules into living cells are vital for probing the physical properties and biochemical interactions that govern the cell's behavior. Here we report the development of a nanoscale cell injection system-termed the nanoinjector-that uses carbon nanotubes to deliver cargo into cells. A single multi-walled carbon nanotube attached to an atomic force microscope tip was functionalized with cargo via a disulfide-based linker. Penetration of cell membranes with this 'nanoneedle', followed by reductive cleavage of the disulfide bonds within the cell's interior, resulted in the release of cargo inside the cells. The capability of the nanoinjector was demonstrated by injection of protein-coated quantum dots into live human cells. Single-particle tracking was employed to characterize the diffusion dynamics of injected quantum dots in the cytosol. This new technique causes no discernible membrane or cell damage, and can deliver a discrete number of molecules to the cell's interior without the requirement of a carrier solvent.

  1. The enhancement of polysulfide absorbsion in Lisbnd S batteries by hierarchically porous CoS2/carbon paper interlayer

    Science.gov (United States)

    Ma, Zhaoling; Li, Zhen; Hu, Kui; Liu, Dongdong; Huo, Jia; Wang, Shuangyin

    2016-09-01

    The high-energy Lisbnd S battery suffers from poor cycling performance due to the shuttle effect of the polysulfides. Strategies must be adopted to suppress the diffusion of polysulfides into the electrolyte in Lisbnd S battery. In this work, for the first time, we adopt hydrophilic carbon paper anchored by hierarchically porous cobalt disulfides as the interlayer for capturing polysulfides through physical absorption and chemical bonding. Hierarchical pores can physically adsorb polysulfides, and moreover cobalt disulfide can trap the polysulfides by forming strong chemical interaction. The sulfur-graphene composite with a sulfur content of 70.5% delivers a high initial capacity of 1239.5 mAh g-1 at 0.2 C and retains a reversible capacity of 818 mAh g-1 after 200 cycles. In spite of a little capacity contribution by the insertion of lithium ions into cobalt disulfide for the initial cycles, it disappears in the subsequent cycling. Therefore, the as-developed porous transition metal disulfides on carbon paper as the interlayer could significantly enhance the cycling performance of Lisbnd S batteries.

  2. Adsorption of odorous sulfur compounds onto activated carbons modified by gamma irradiation.

    Science.gov (United States)

    Vega, Esther; Sánchez-Polo, Manuel; Gonzalez-Olmos, Rafael; Martin, María J

    2015-11-01

    A commercial activated carbon (AC) was modified by gamma irradiation and was tested as adsorbent for the removal of ethyl mercaptan, dimethyl disulfide and dimethyl disulfide in wet conditions. Modifications were carried out under five different conditions: irradiation in absence of water, in presence of ultrapure water, in ultrapure water at pH=1.0 and 1000 mg L(-1) Cl(-), in ultrapure water at pH=7.5 and 1000 mg L(-1) Br(-), and in ultrapure water at pH=12.5 and 1000 mg L(-1) NO3(-). The chemical properties of each AC were characterized by elemental analysis, temperature programmed desorption and X-ray photoelectron spectroscopy. Outcomes show that a large number of oxygen functional groups were incorporated in the AC surface by gamma irradiation, especially in the AC irradiated in the presence of ultrapure water. The dynamic adsorption test results reveal that the incorporation of oxygen functional groups did not enhance the adsorption capacities for dimethyl sulfide and dimethyl disulfide. A significant improvement in the ethyl mercaptan adsorption capacity was correlated with the incorporation of phenolic groups in the AC surface. Moreover, diethyl disulfide was detected as by-product of ethyl mercaptan oxidation process under wet conditions and its formation depended on the chemical properties of ACs. PMID:26160734

  3. Identification of a Disulfide Bridge in Sodium-Coupled Neutral Amino Acid Transporter 2(SNAT2) by Chemical Modification.

    Science.gov (United States)

    Chen, Chen; Wang, Jiahong; Cai, Ruiping; Yuan, Yanmeng; Guo, Zhanyun; Grewer, Christof; Zhang, Zhou

    2016-01-01

    Sodium-coupled neutral amino acid transporter 2 (SNAT2) belongs to solute carrier 38 (SLC38) family of transporters, which is ubiquitously expressed in mammalian tissues and mediates transport of small, neutral amino acids, exemplified by alanine(Ala, A). Yet structural data on SNAT2, including the relevance of intrinsic cysteine residues on structure and function, is scarce, in spite of its essential roles in many tissues. To better define the potential of intrinsic cysteines to form disulfide bonds in SNAT2, mutagenesis experiments and thiol-specific chemical modifications by N-ethylmaleimide (NEM) and methoxy-polyethylene glycol maleimide (mPEG-Mal, MW 5000) were performed, with or without the reducing regent dithiothreitol (DTT) treatment. Seven single mutant transporters with various cysteine (Cys, C) to alanine (Ala, A) substitutions, and a C245,279A double mutant were introduced to SNAT2 with a hemagglutinin (HA) tag at the C-terminus. The results showed that the cells expressing C245A or C279A were labeled by one equivalent of mPEG-Mal in the presence of DTT, while wild-type or all the other single Cys to Ala mutants were modified by two equivalents of mPEG-Mal. Furthermore, the molecular weight of C245,279A was not changed in the presence or absence of DTT treatment. The results suggest a disulfide bond between Cys245 and Cys279 in SNAT2 which has no effect on cell surface trafficking, as well as transporter function. The proposed disulfide bond may be important to delineate proximity in the extracellular domain of SNAT2 and related proteins. PMID:27355203

  4. Enhancing the thermostability of a cold-active lipase from Penicillium cyclopium by in silico design of a disulfide bridge.

    Science.gov (United States)

    Tan, Zhongbiao; Li, Jianfang; Wu, Minchen; Wang, Junqing

    2014-08-01

    Cysteine mutants of a cold-active lipase (PcLipI) from Penicillium cyclopium were designed by the software Disulfide by Design Ver. 1.20 in an effort to improve enzyme thermostability by addition of a disulfide bridge. Those mutants predicted by molecular dynamics simulation to have better thermostability than the wild type were first expressed in Escherichia coli BL21(DE3) and then, for further investigation, in Pichia pastoris GS115. By replacing Val248 and Thr251 with cysteines to create a disulfide bridge, the recombinant lipases reE-PcLipV248C-T251C (expressed in E. coli) and reP-PcLipV248C-T251C (expressed in P. pastoris) were obtained. Both had enhanced thermostability with half-lives at 35 °C about 4.5- and 12.8-fold longer than that of the parent PcLipI expressed in E. coli and P. pastoris, respectively. The temperature optima of reE-PcLipV248C-T251C and reP-PcLipV248C-T251C were 35 and 30 °C, which were each 5 °C higher than those of the parent PcLipI expressed in E. coli and P. pastoris. The K ms of reE-PcLipV248C-T251C and reP-PcLipV248C-T251C toward tributyrin were 53.2 and 39.5 mM, while their V maxs were 1,460 and 3,800 U/mg, respectively. PcLipV248C-T251C had better thermostability and catalytic efficiency than the other mutants and the parent PcLipI. PMID:24867629

  5. Mixed disulfide formation at Cys141 leads to apparent unidirectional attenuation of Aspergillus niger NADP-glutamate dehydrogenase activity.

    Directory of Open Access Journals (Sweden)

    Adhish S Walvekar

    Full Text Available NADP-Glutamate dehydrogenase from Aspergillus niger (AnGDH exhibits sigmoid 2-oxoglutarate saturation. Incubation with 2-hydroxyethyl disulfide (2-HED, the disulfide of 2-mercaptoethanol resulted in preferential attenuation of AnGDH reductive amination (forward activity but with a negligible effect on oxidative deamination (reverse activity, when monitored in the described standard assay. Such a disulfide modified AnGDH displaying less than 1.0% forward reaction rate could be isolated after 2-HED treatment. This unique forward inhibited GDH form (FIGDH, resembling a hypothetical 'one-way' active enzyme, was characterized. Kinetics of 2-HED mediated inhibition and protein thiol titrations suggested that a single thiol group is modified in FIGDH. Two site-directed cysteine mutants, C141S and C415S, were constructed to identify the relevant thiol in FIGDH. The forward activity of C141S alone was insensitive to 2-HED, implicating Cys141 in FIGDH formation. It was observed that FIGDH displayed maximal reaction rate only after a pre-incubation with 2-oxoglutarate and NADPH. In addition, compared to the native enzyme, FIGDH showed a four fold increase in K0.5 for 2-oxoglutarate and a two fold increase in the Michaelis constants for ammonium and NADPH. With no change in the GDH reaction equilibrium constant, the FIGDH catalyzed rate of approach to equilibrium from reductive amination side was sluggish. Altered kinetic properties of FIGDH at least partly account for the observed apparent loss of forward activity when monitored under defined assay conditions. In sum, although Cys141 is catalytically not essential, its covalent modification provides a striking example of converting the biosynthetic AnGDH into a catabolic enzyme.

  6. Oxidation of p53 through DNA charge transport involves a network of disulfides within the DNA-binding domain.

    Science.gov (United States)

    Schaefer, Kathryn N; Geil, Wendy M; Sweredoski, Michael J; Moradian, Annie; Hess, Sonja; Barton, Jacqueline K

    2015-01-27

    Transcription factor p53 plays a critical role in the cellular response to stress stimuli. We have seen that p53 dissociates selectively from various promoter sites as a result of oxidation at long-range through DNA-mediated charge transport (CT). Here, we examine this chemical oxidation and determine the residues in p53 that are essential for oxidative dissociation, focusing on the network of cysteine residues adjacent to the DNA-binding site. Of the eight mutants studied, only the C275S mutation shows decreased affinity for the Gadd45 promoter site. However, both mutations C275S and C277S result in substantial attenuation of oxidative dissociation, with C275S causing the most severe attenuation. Differential thiol labeling was used to determine the oxidation states of cysteine residues within p53 after DNA-mediated oxidation. Reduced cysteines were iodoacetamide-labeled, whereas oxidized cysteines participating in disulfide bonds were (13)C2D2-iodoacetamide-labeled. Intensities of respective iodoacetamide-modified peptide fragments were analyzed by mass spectrometry. A distinct shift in peptide labeling toward (13)C2D2-iodoacetamide-labeled cysteines is observed in oxidized samples, confirming that chemical oxidation of p53 occurs at long range. All observable cysteine residues trend toward the heavy label under conditions of DNA CT, indicating the formation of multiple disulfide bonds among the cysteine network. On the basis of these data, it is proposed that disulfide formation involving C275 is critical for inducing oxidative dissociation of p53 from DNA. PMID:25584637

  7. Role of cysteine-58 and cysteine-95 residues in the thiol di-sulfide oxidoreductase activity of Macrophage Migration Inhibitory Factor-2 of Wuchereria bancrofti.

    Science.gov (United States)

    Chauhan, Nikhil; Hoti, S L

    2016-01-01

    Macrophage Migration Inhibitory Factor (MIF) is the first human cytokine reported and was thought to have a central role in the regulation of inflammatory responses. Homologs of this molecule have been reported in bacteria, invertebrates and plants. Apart from cytokine activity, it also has two catalytic activities viz., tautomerase and di-sulfide oxidoreductase, which appear to be involved in immunological functions. The CXXC catalytic site is responsible for di-sulfide oxidoreductase activity of MIF. We have recently reported thiol-disulfide oxidoreductase activity of Macrophage Migration Inhibitory Factor-2 of Wuchereria bancrofti (Wba-MIF-2), although it lacks the CXXC motif. We hypothesized that three conserved cysteine residues might be involved in the formation of di-sulfide oxidoreductase catalytic site. Homology modeling of Wba-MIF-2 showed that among the three cysteine residues, Cys58 and Cys95 residues came in close proximity (3.23Å) in the tertiary structure with pKa value 9, indicating that these residues might play a role in the di-sulfide oxidoreductase catalytic activity. We carried out site directed mutagenesis of these residues (Cys58Ser & Cys95Ser) and expressed mutant proteins in Escherichia coli. The mutant proteins did not show any oxidoreductase activity in the insulin reduction assay, thus indicating that these two cysteine residues are vital for the catalytic activity of Wba-MIF-2. PMID:26432350

  8. Crystal Structure of Reduced and of Oxidized Peroxiredoxin IV Enzyme Reveals a Stable Oxidized Decamer and a Non-disulfide-bonded Intermediate in the Catalytic Cycle*

    Science.gov (United States)

    Cao, Zhenbo; Tavender, Timothy J.; Roszak, Aleksander W.; Cogdell, Richard J.; Bulleid, Neil J.

    2011-01-01

    Peroxiredoxin IV (PrxIV) is an endoplasmic reticulum-localized enzyme that metabolizes the hydrogen peroxide produced by endoplasmic reticulum oxidase 1 (Ero1). It has been shown to play a role in de novo disulfide formation, oxidizing members of the protein disulfide isomerase family of enzymes, and is a member of the typical 2-Cys peroxiredoxin family. We have determined the crystal structure of both reduced and disulfide-bonded, as well as a resolving cysteine mutant of human PrxIV. We show that PrxIV has a similar structure to other typical 2-Cys peroxiredoxins and undergoes a conformational change from a fully folded to a locally unfolded form following the formation of a disulfide between the peroxidatic and resolving cysteine residues. Unlike other mammalian typical 2-Cys peroxiredoxins, we show that human PrxIV forms a stable decameric structure even in its disulfide-bonded state. In addition, the structure of a resolving cysteine mutant reveals an intermediate in the reaction cycle that adopts the locally unfolded conformation. Interestingly the peroxidatic cysteine in the crystal structure is sulfenylated rather than sulfinylated or sulfonylated. In addition, the peroxidatic cysteine in the resolving cysteine mutant is resistant to hyper-oxidation following incubation with high concentrations of hydrogen peroxide. These results highlight some unique properties of PrxIV and suggest that the equilibrium between the fully folded and locally unfolded forms favors the locally unfolded conformation upon sulfenylation of the peroxidatic cysteine residue. PMID:21994946

  9. The road to the first, fully active and more stable human insulin variant with an additional disulfide bond

    DEFF Research Database (Denmark)

    Vinther, Tine N.; Kjeldsen, Thomas B.; Jensen, Knud Jørgen;

    2015-01-01

    variants in vertebrates consist of two peptide chains and have six cysteine residues, which form three disulfide bonds, two of them link the two chains and a third is an intra-chain bond in the A-chain. This classical insulin fold appears to have been conserved over half a billion years of evolution. We...... with markedly improved stability and gained insights into the instability of analogs with seven cysteine residues, importance of dimerization for stability, insulin fibril formation process, and the conformation of insulin binding to its receptor. Our results also open the way for new strategies...

  10. Processing, disulfide pattern, and biological activity of a sugar beet defensin, AX2, expressed in Pichia pastoris

    DEFF Research Database (Denmark)

    Kristensen, A K; Brunstedt, J; Nielsen, J E;

    1999-01-01

    activity of the recombinant protein were determined and compared with that of the authentic AX2. In P. pastoris, the protein was expressed with an additional N-terminal arginine. The disulfide bonding was found to be identical to that of the authentic AX2. However, when tested in in vitro bioassay......, the biological activity of the recombinant protein was slightly lower than that measured for the authentic protein. Furthermore, the recombinant protein was significantly more sensitive to Ca(2+) than the authentic protein. This is most probably due to the extra arginine, since no other differences between...

  11. Urea- Hydrogen Peroxide (UHP Oxidation of Thiols to the Corresponding Disulfides Promoted by Maleic Anhydride as Mediator

    Directory of Open Access Journals (Sweden)

    M. H. Habibi

    2005-10-01

    Full Text Available Urea-hydrogen peroxide (UHP was used in the presence of maleic anhydride as mediator in a simple and convenient method for the oxidation in high yield of some thiols to the corresponding disulfides. Peroxymaleic acid formed in situ from the reaction of UHP with maleic anhydride has a key role in this oxidation. Performance of the reaction in various solvents showed that methanol was the solvent of choice at 0 oC. The products were isolated by simple filtration on silica gel.

  12. Urea- Hydrogen Peroxide (UHP) Oxidation of Thiols to the Corresponding Disulfides Promoted by Maleic Anhydride as Mediator

    OpenAIRE

    M. H. Habibi; M. Montazerozohori; Bahador Karami

    2005-01-01

    Urea-hydrogen peroxide (UHP) was used in the presence of maleic anhydride as mediator in a simple and convenient method for the oxidation in high yield of some thiols to the corresponding disulfides. Peroxymaleic acid formed in situ from the reaction of UHP with maleic anhydride has a key role in this oxidation. Performance of the reaction in various solvents showed that methanol was the solvent of choice at 0 oC. The products were isolated by simple filtration on silica gel.

  13. A Periplasmic LolA Derivative with a Lethal Disulfide Bond Activates the Cpx Stress Response System▿

    OpenAIRE

    Tao, Kazuyuki; Watanabe, Shoji; Narita, Shin-ichiro; Tokuda, Hajime

    2010-01-01

    LolA accommodates the acyl chains of lipoproteins in its hydrophobic cavity and shuttles between the inner and outer membranes through the hydrophilic periplasm to place lipoproteins in the outer membrane. The LolA(I93C/F140C) derivative, in which Cys replaces Ile at position 93 and Phe at position 140, strongly inhibited growth in the absence of a reducing agent because of the lethal intramolecular disulfide bond between the two Cys residues. Expression of I93C/F140C was found to activate th...

  14. Disulfide bond formation and folding of plant peroxidases expressed as inclusion body protein in Escherichia coli thioredoxin reductase negative strains

    DEFF Research Database (Denmark)

    Teilum, K; Ostergaard, L; Welinder, K G

    1999-01-01

    Escherichia coli is widely used for the production of proteins, which are of interest in structure and function studies. The folding yield of inclusion body protein is, however, generally low (a few percent) for proteins such as the plant and fungal peroxidases, which contain four disulfide bonds......, two Ca2+ ions, and a heme group. We have studied the expression yield and folding efficiency of (i) a novel Arabidopsis thaliana peroxidase, ATP N; and (ii) barley grain peroxidase, BP 1. The expression yield ranges from 0 to 60 microgram/ml of cell culture depending on the peroxidase gene and the...

  15. Toxoplasma gondii protein disulfide isomerase (TgPDI is a novel vaccine candidate against toxoplasmosis.

    Directory of Open Access Journals (Sweden)

    Hai-Long Wang

    Full Text Available Toxoplasma gondii is a ubiquitous protozoan parasite that can infect all warm-blooded animals, including both mammals and birds. Protein disulfide isomerase (PDI localises to the surface of T. gondii tachyzoites and modulates the interactions between parasite and host cells. In this study, the protective efficacy of recombinant T. gondii PDI (rTgPDI as a vaccine candidate against T. gondii infection in BALB/c mice was evaluated. rTgPDI was expressed and purified from Escherichia coli. Five groups of animals (10 animals/group were immunised with 10, 20, 30, 40 μg of rTgPDI per mouse or with PBS as a control group. All immunisations were performed via the nasal route at 1, 14 and 21 days. Two weeks after the last immunisation, the immune responses were evaluated by lymphoproliferative assays and by cytokine and antibody measurements. The immunised mice were challenged with tachyzoites of the virulent T. gondii RH strain on the 14th day after the last immunisation. Following the challenge, the tachyzoite loads in tissues were assessed, and animal survival time was recorded. Our results showed that the group immunised with 30 μg rTgPDI showed significantly higher levels of specific antibodies against the recombinant protein, a strong lymphoproliferative response and significantly higher levels of IgG2a, IFN-gamma (IFN-γ, IL-2 and IL-4 production compared with other doses and control groups. While no changes in IL-10 levels were detected. After being challenged with T. gondii tachyzoites, the numbers of tachyzoites in brain and liver tissues from the rTgPDI group were significantly reduced compared with those of the control group, and the survival time of the mice in the rTgPDI group was longer than that of mice in the control group. Our results showed that immunisation with rTgPDI elicited a protective immune reaction and suggested that rTgPDI might represent a promising vaccine candidate for combating toxoplasmosis.

  16. Protein disulfide isomerase interacts with tau protein and inhibits its fibrillization.

    Directory of Open Access Journals (Sweden)

    Li-Rong Xu

    Full Text Available BACKGROUND: Tau protein is implicated in the pathogenesis of neurodegenerative disorders such as tauopathies including Alzheimer disease, and Tau fibrillization is thought to be related to neuronal toxicity. Physiological inhibitors of Tau fibrillization hold promise for developing new strategies for treatment of Alzheimer disease. Because protein disulfide isomerase (PDI is both an enzyme and a chaperone, and implicated in neuroprotection against Alzheimer disease, we want to know whether PDI can prevent Tau fibrillization. In this study, we have investigated the interaction between PDI and Tau protein and the effect of PDI on Tau fibrillization. METHODOLOGY/PRINCIPAL FINDINGS: As evidenced by co-immunoprecipitation and confocal laser scanning microscopy, human PDI interacts and co-locates with some endogenous human Tau on the endoplasmic reticulum of undifferentiated SH-SY5Y neuroblastoma cells. The results from isothermal titration calorimetry show that one full-length human PDI binds to one full-length human Tau (or human Tau fragment Tau244-372 monomer with moderate, micromolar affinity at physiological pH and near physiological ionic strength. As revealed by thioflavin T binding assays, Sarkosyl-insoluble SDS-PAGE, and transmission electron microscopy, full-length human PDI remarkably inhibits both steps of nucleation and elongation of Tau244-372 fibrillization in a concentration-dependent manner. Furthermore, we find that two molecules of the a-domain of human PDI interact with one Tau244-372 molecule with sub-micromolar affinity, and inhibit both steps of nucleation and elongation of Tau244-372 fibrillization more strongly than full-length human PDI. CONCLUSIONS/SIGNIFICANCE: We demonstrate for the first time that human PDI binds to Tau protein mainly through its thioredoxin-like catalytic domain a, forming a 1∶1 complex and preventing Tau misfolding. Our findings suggest that PDI could act as a physiological inhibitor of Tau

  17. Massilia putida sp. nov., a dimethyl disulfide-producing bacterium isolated from wolfram mine tailing.

    Science.gov (United States)

    Feng, Guang-Da; Yang, Song-Zhen; Li, Hua-Ping; Zhu, Hong-Hui

    2016-01-01

    A heavy metal-resistant and dimethyl disulfide-producing bacterial strain, designated 6NM-7T, was isolated from wolfram mine tailing, Dayu County, Jiangxi Province, PR China. Strain 6NM-7T was aerobic, Gram-stain-negative and motile by means of a single polar flagellum. Phylogenetic analysis, based on 16S rRNA gene sequences, showed that strain 6NM-7T was affiliated with the genus Massilia and was closely related to Massilia norwichensis LMG 28164T (98.8 % 16S rRNA gene sequence similarity), Massilia kyonggiensis KACC 17471T (98.4 %), Massilia niastensis KACC 12599T (97.8 %), Massilia tieshanensis KACC 14940T (97.3 %), Massilia haematophila KACC 13771T (97.2 %), Massilia namucuonensis CGMCC 1.11014T (97.1 %) and Massilia aerilata KACC 12505T (97.1 %). The DNA-DNA relatedness values between strain 6NM-7T and its closely related type strains were all below 70 %. The major respiratory quinone was unbiquinone 8 (Q-8) and the major cellular fatty acids consisted of C16 : 0 (33.2 %), summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH; 21.8 %), C17 : 0 cyclo (20.8 %), C18 : 1ω7c (7.4 %) and C10 : 0 3-OH (5.8 %). The major polar lipids consisted of phosphatidylethanolamine, phosphatidylglycerol and diphosphatidylglycerol. The genomic DNA G+C content of strain 6NM-7T was 66.8 ± 0.6 mol%. On the basis of the results of this polyphasic taxonomic study, strain 6NM-7T should be assigned to a novel species of the genus Massilia, for which the name Massilia putida sp. nov. is proposed. The type strain is 6NM-7T ( = DSM 27523T = KCTC 42761T). PMID:26449383

  18. Structures of cGMP-Dependent Protein Kinase (PKG) Iα Leucine Zippers Reveal an Interchain Disulfide Bond Important for Dimer Stability

    Science.gov (United States)

    Qin, Liying; Reger, Albert S.; Guo, Elaine; Yang, Matthew P.; Zwart, Peter; Casteel, Darren E.; Kim, Choel

    2016-01-01

    cGMP-dependent protein kinase (PKG) Iα is a central regulator of smooth muscle tone and vasorelaxation. The N-terminal leucine zipper (LZ) domain dimerizes and targets PKG Iα by interacting with G-kinase-anchoring proteins. The PKG Iα LZ contains C42 that is known to form a disulfide bond upon oxidation and to activate PKG Iα. To understand the molecular details of the PKG Iα LZ and C42–C42′ disulfide bond, we determined crystal structures of the PKG Iα wild-type (WT) LZ and C42L LZ. Our data demonstrate that the C42–C42′ disulfide bond dramatically stabilizes PKG Iα and that the C42L mutant mimics the oxidized WT LZ structurally. PMID:26132214

  19. Carbon Carbon Composites: An Overview .

    OpenAIRE

    G. Rohini Devi; K. Rama Rao

    1993-01-01

    Carbon carbon composites are a new class of engineering materials that are ceramic in nature but exhibit brittle to pseudoplastic behaviour. Carbon-carbon is a unique all-carbon composite with carbon fibre embeded in carbon matrix and is known as an inverse composite. Due to their excellent thermo-structural properties, carbon-carbon composites are used in specialised application like re-entry nose-tips, leading edges, rocket nozzles, and aircraft brake discs apart from several indust...

  20. Loss of metal ions, disulfide reduction and mutations related to familial ALS promote formation of amyloid-like aggregates from superoxide dismutase.

    Directory of Open Access Journals (Sweden)

    Zeynep A Oztug Durer

    Full Text Available Mutations in the gene encoding Cu-Zn superoxide dismutase (SOD1 are one of the causes of familial amyotrophic lateral sclerosis (FALS. Fibrillar inclusions containing SOD1 and SOD1 inclusions that bind the amyloid-specific dye thioflavin S have been found in neurons of transgenic mice expressing mutant SOD1. Therefore, the formation of amyloid fibrils from human SOD1 was investigated. When agitated at acidic pH in the presence of low concentrations of guanidine or acetonitrile, metalated SOD1 formed fibrillar material which bound both thioflavin T and Congo red and had circular dichroism and infrared spectra characteristic of amyloid. While metalated SOD1 did not form amyloid-like aggregates at neutral pH, either removing metals from SOD1 with its intramolecular disulfide bond intact or reducing the intramolecular disulfide bond of metalated SOD1 was sufficient to promote formation of these aggregates. SOD1 formed amyloid-like aggregates both with and without intermolecular disulfide bonds, depending on the incubation conditions, and a mutant SOD1 lacking free sulfhydryl groups (AS-SOD1 formed amyloid-like aggregates at neutral pH under reducing conditions. ALS mutations enhanced the ability of disulfide-reduced SOD1 to form amyloid-like aggregates, and apo-AS-SOD1 formed amyloid-like aggregates at pH 7 only when an ALS mutation was also present. These results indicate that some mutations related to ALS promote formation of amyloid-like aggregates by facilitating the loss of metals and/or by making the intramolecular disulfide bond more susceptible to reduction, thus allowing the conversion of SOD1 to a form that aggregates to form resembling amyloid. Furthermore, the occurrence of amyloid-like aggregates per se does not depend on forming intermolecular disulfide bonds, and multiple forms of such aggregates can be produced from SOD1.

  1. Molecular Cloning, Expression Analysis, and Preliminarily Functional Characterization of the Gene Encoding Protein Disulfide Isomerase from Jatropha curcas.

    Science.gov (United States)

    Wang, Haibo; Zou, Zhurong; Gong, Ming

    2015-05-01

    Reactive oxygen species (ROS) in plants, arising from various environmental stresses, impair the thiol-contained proteins that are susceptible to irregular oxidative formation of disulfide bonds, which might be alleviated by a relatively specific modifier called protein disulfide isomerase (PDI). From our previous data of the transcriptome and digital gene expression of cold-hardened Jatropha curcas, a PDI gene was proposed to be cold-relevant. In this study, its full-length cDNA (JcPDI) was cloned, with the size of 1649 bp containing the entire open reading frame (ORF) of 1515 bp. This ORF encodes a polypeptide of 504 amino acids with theoretical molecular weight of 56.6 kDa and pI value of 4.85. One N-terminal signal peptide (-MASKGSIWSCMFLFSLI VAISAGEG-) and the C-terminal anchoring sequence motif (-KDEL-) specific to the endoplasmic reticulum, as well as two thioredoxin domains (-CGHC-), are also found by predictions. Through semi-quantitative RT-PCR, the expression of JcPDI was characterized to be tissue-differential strongly in leaves and roots, but weakly in stems, and of cold-induced alternations. Furthermore, JcPDI overexpression in yeast could notably enhance the cold resistance of host cells. Conclusively, these results explicitly suggested a considerable association of JcPDI to cold response and a putative application potential for its correlated genetic engineering. PMID:25825250

  2. Sample limited characterization of a novel disulfide-rich venom peptide toxin from terebrid marine snail Terebra variegata.

    Directory of Open Access Journals (Sweden)

    Prachi Anand

    Full Text Available Disulfide-rich peptide toxins found in the secretions of venomous organisms such as snakes, spiders, scorpions, leeches, and marine snails are highly efficient and effective tools for novel therapeutic drug development. Venom peptide toxins have been used extensively to characterize ion channels in the nervous system and platelet aggregation in haemostatic systems. A significant hurdle in characterizing disulfide-rich peptide toxins from venomous animals is obtaining significant quantities needed for sequence and structural analyses. Presented here is a strategy for the structural characterization of venom peptide toxins from sample limited (4 ng specimens via direct mass spectrometry sequencing, chemical synthesis and NMR structure elucidation. Using this integrated approach, venom peptide Tv1 from Terebra variegata was discovered. Tv1 displays a unique fold not witnessed in prior snail neuropeptides. The novel structural features found for Tv1 suggest that the terebrid pool of peptide toxins may target different neuronal agents with varying specificities compared to previously characterized snail neuropeptides.

  3. Isolation, Characterization, and Synthesis of the Barrettides: Disulfide-Containing Peptides from the Marine Sponge Geodia barretti.

    Science.gov (United States)

    Carstens, Bodil B; Rosengren, K Johan; Gunasekera, Sunithi; Schempp, Stefanie; Bohlin, Lars; Dahlström, Mia; Clark, Richard J; Göransson, Ulf

    2015-08-28

    Two disulfide-containing peptides, barrettides A (1) and B (2), from the cold-water marine sponge Geodia barretti are described. Those 31 amino acid residue long peptides were sequenced using mass spectrometry methods and structurally characterized using NMR spectroscopy. The structure of 1 was confirmed by total synthesis using the solid-phase peptide synthesis approach that was developed. The two peptides were found to differ only at a single position in their sequence. The three-dimensional structure of 1 revealed that these peptides possess a unique fold consisting of a long β-hairpin structure that is cross-braced by two disulfide bonds in a ladder-like arrangement. The peptides are amphipathic in nature with the hydrophobic and charged residues clustered on separate faces of the molecule. The barrettides were found not to inhibit the growth of either Escherichia coli or Staphylococcus aureus but displayed antifouling activity against barnacle larvae (Balanus improvisus) without lethal effects in the concentrations tested. PMID:26222779

  4. Micelle-Vesicle Transition by Cleavage of Disulfide Spacer Chain for Gemini Surfactant in Didodecyldimethylammonium Chloride Aqueous Solutions.

    Science.gov (United States)

    Mizuhashi, Toshinari; Asakawa, Tsuyoshi; Ohta, Akio

    2015-01-01

    We examined the "micelle-vesicle transition" through the mixing effect of single-tailed thiol surfactants produced by the cleavage of gemini surfactants, [C12H25N(CH3)2CH2CH2SSCH2CH2N(CH3)2C12H25] 2Cl (C12SSC12), which have a disulfide bond in the spacer chain. Phase diagrams of C12H25N(CH3)2CH2CH2SHCl-didodecyldimethylammonium chloride (C12SH-DDAC) and C12SSC12-DDAC were determined by conductivity and pyrene fluorescence probe methods. The aggregate diameters were evaluated by dynamic light scattering (DLS). The critical vesicle concentration (CVC) was confirmed by the abrupt increase in the intensity of light scattering with excitation at 335 nm. Vesicle formation was confined to the DDAC-rich region of the C12SSC12-DDAC system, while the vesicle formation region for the C12SH-DDAC system spread out with the addition of dithiothreitol (DTT) to C12SSC12-DDAC. This implies that single-tailed surfactants can induce a more favorable environment for molecular packing of the vesicular surface. The micelle-vesicle transition occurs with disulfide spacer chain cleavage of gemini surfactants at a particular specific concentration range. PMID:26250426

  5. The Interplay of Disulfide Bonds, α-Helicity, and Hydrophobic Interactions Leads to Ultrahigh Proteolytic Stability of Peptides.

    Science.gov (United States)

    Chen, Yaqi; Yang, Chaoqiong; Li, Tao; Zhang, Miao; Liu, Yang; Gauthier, Marc A; Zhao, Yibing; Wu, Chuanliu

    2015-08-10

    The contribution of noncovalent interactions to the stability of naturally occurring peptides and proteins has been generally acknowledged, though how these can be rationally manipulated to improve the proteolytic stability of synthetic peptides remains to be explored. In this study, a platform to enhance the proteolytic stability of peptides was developed by controllably dimerizing them into α-helical dimers, connected by two disulfide bonds. This platform not only directs peptides toward an α-helical conformation but permits control of the interfacial hydrophobic interactions between the peptides of the dimer. Using two model dimeric systems constructed from the N-terminal α-helix of RNase A and known inhibitors for the E3 ubiquitin ligase MDM2 (and its homologue MDMX), a deeper understanding into the interplay of disulfide bonds, α-helicity, and hydrophobic interactions on enhanced proteolytic stability was sought out. Results reveal that all three parameters play an important role on attaining ultrahigh proteolytic resistance, a concept that can be exploited for the development of future peptide therapeutics. The understanding gained through this study will enable this strategy to be tailored to new peptides because the proposed strategy displays substantial tolerance to sequence permutation. It thus appears promising for conveniently creating prodrugs composed entirely of the therapeutic peptide itself (i.e., in the form of a dimer).

  6. Role of disulfide linkages in desulfurization chemistry. The reactions of benzenethiol on a sulfur-covered Mo(110) surface

    Energy Technology Data Exchange (ETDEWEB)

    Weldon, M.K.; Napier, M.E.; Wiegand, B.C.; Friend, C.M. [Harvard Univ., Cambridge, MA (United States); Uvdal, P. [Lund Univ. (Sweden)

    1994-09-07

    The reactions of benzenethiol on a sulfur-covered Mo(110) surface were studied using temperature programmed reaction, X-ray photoelectron, and high resolution electron energy loss spectroscopies. The sulfur overlayer profoundly alters the kinetics and selectivity for desulfurization and dehydrogenation. By using isotopic labeling, we have established that phenyl disulfide (C{sub 6}H{sub 5}S-S-) is formed via S-H bond scission and S-S bond formation on Mo(110) at 100 K. The S-S- linkage is oriented perpendicular and the phenyl ring parallel to the surface. The disulfide subsequently forms an upright phenylthiolate species, bound directly to the Mo(110) surface, prior to the onset of benzene formation at 300 K. In contrast to the clean surface, where only the low-temperature state is observed, a second benzene peak is observed at 500 K on the sulfur-covered surface. This feature is attributed to disproportionation of surface phenyl groups to produce gaseous benzene and surface benzyne. In addition, gaseous phenyl also desorbs from the surface in the same temperature range, due to a lack of available surface hydrogen. The selectivity for gaseous hydrocarbon production is approximately 80%, nearly twice that on the clean surface, while the total amount of reaction remains the same. 40 refs., 5 figs., 2 tabs.

  7. Im10A, a short conopeptide isolated from Conus imperialis and possesses two highly concentrated disulfide bridges and analgesic activity.

    Science.gov (United States)

    Yu, Shuo; Du, Tianpeng; Liu, Zhuguo; Wu, Qiaoling; Feng, Guixue; Dong, Mingxin; Zhou, Xiaowei; Jiang, Ling; Dai, Qiuyun

    2016-07-01

    In the present study, we isolated, synthesized and NMR structurally characterized a novel conopeptide Im10A consisting of 11 amino acids (NTICCEGCMCY-NH2) from Conus imperialis. Unlike other conopeptides with four cysteine residues, Im10A had only two residues in loop 1 and one residue in loop 2 (CC-loop1-C-loop2-C), which formed a stable disulfide connectivity "I-IV, II- III" (framework X) with a type I β-turn. Interestingly, Im10A exhibited 50.7% analgesic activity on rat partial sciatic nerve ligation (PNL) at 2h after Im10A administration. However, 10μM Im10A exhibited no apparent effect on neuronal nicotinic acetylcholine receptor, and it did not target DRG voltage-dependent sodium, potassium and calcium ion channels and opioid receptor. To our knowledge, Im10A had the most concentrated disulfide bridges among conopeptides with four cysteine residues. This finding provided a new motif for the future development of biomimetic compounds. PMID:27131596

  8. Effect of pharmaceutical potential endocrine disruptor compounds on protein disulfide isomerase reductase activity using di-eosin-oxidized-glutathione.

    Directory of Open Access Journals (Sweden)

    Danièle Klett

    Full Text Available BACKGROUND: Protein Disulfide Isomerase (PDI in the endoplasmic reticulum of all cells catalyzes the rearrangement of disulfide bridges during folding of membrane and secreted proteins. As PDI is also known to bind various molecules including hormones such as estradiol and thyroxin, we considered the hypothesis that adverse effects of endocrine-disrupter compounds (EDC could be mediated through their interaction with PDI leading to defects in membrane or secreted proteins. METHODOLOGY/PRINCIPAL FINDINGS: Taking advantage of the recent description of the fluorescence self quenched substrate di-eosin-oxidized-glutathione (DiE-GSSG, we determined kinetically the effects of various potential pharmaceutical EDCs on the in-vitro reductase activity of bovine liver PDI by measuring the fluorescence of the reaction product (E-GSH. Our data show that estrogens (ethynylestradiol and bisphenol-A as well as indomethacin exert an inhibition whereas medroxyprogesteroneacetate and nortestosterone exert a potentiation of bovine PDI reductase activity. CONCLUSIONS: The present data indicate that the tested EDCs could not only affect endocrine target cells through nuclear receptors as previously shown, but could also affect these and all other cells by positively or negatively affecting PDI activity. The substrate DiE-GSSG has been demonstrated to be a convenient substrate to measure PDI reductase activity in the presence of various potential EDCs. It will certainly be usefull for the screening of potential effect of all kinds of chemicals on PDI reductase activity.

  9. Preliminary crystallographic data of the three homologues of the thiol–disulfide oxidoreductase DsbA in Neisseria meningitidis

    International Nuclear Information System (INIS)

    The Neisseria meningitidis genome possesses three genes encoding active DsbAs. To throw light on the reason for this genetic multiplicity, the three enzymes have been purified and crystallized. Bacterial virulence depends on the correct folding of surface-exposed proteins, a process that is catalyzed by the thiol-disulfide oxidoreductase DsbA, which facilitates the synthesis of disulfide bonds in Gram-negative bacteria. Uniquely among bacteria, the Neisseria meningitidis genome possesses three genes encoding active DsbAs: DsbA1, DsbA2 and DsbA3. DsbA1 and DsbA2 have been characterized as lipoproteins involved in natural competence and in host-interactive biology, while the function of DsbA3 remains unknown. In an attempt to shed light on the reason for this multiplicity of dsbA genes, the three enzymes from N. meningitidis have been purified and crystallized in the presence of high concentrations of ammonium sulfate. The best crystals were obtained using DsbA1 and DsbA3; they belong to the orthorhombic and tetragonal systems and diffract to 1.5 and 2.7 Å resolution, respectively

  10. Linear array of conserved sequence motifs to discriminate protein subfamilies: study on pyridine nucleotide-disulfide reductases

    Directory of Open Access Journals (Sweden)

    De Las Rivas Javier

    2007-03-01

    Full Text Available Abstract Background The pyridine nucleotide disulfide reductase (PNDR is a large and heterogeneous protein family divided into two classes (I and II, which reflect the divergent evolution of its characteristic disulfide redox active site. However, not all the PNDR members fit into these categories and this suggests the need of further studies to achieve a more comprehensive classification of this complex family. Results A workflow to improve the clusterization of protein families based on the array of linear conserved motifs is designed. The method is applied to the PNDR large family finding two main groups, which correspond to PNDR classes I and II. However, two other separate protein clusters, previously classified as class I in most databases, are outgrouped: the peroxide reductases (NAOX, NAPE and the type II NADH dehydrogenases (NDH-2. In this way, two novel PNDR classes III and IV for NAOX/NAPE and NDH-2 respectively are proposed. By knowledge-driven biochemical and functional data analyses done on the new class IV, a linear array of motifs putatively related to Cu(II-reductase activity is detected in a specific subset of NDH-2. Conclusion The results presented are a novel contribution to the classification of the complex and large PNDR protein family, supporting its reclusterization into four classes. The linear array of motifs detected within the class IV PNDR subfamily could be useful as a signature for a particular subgroup of NDH-2.

  11. Carbon Carbon Composites: An Overview .

    Directory of Open Access Journals (Sweden)

    G. Rohini Devi

    1993-10-01

    Full Text Available Carbon carbon composites are a new class of engineering materials that are ceramic in nature but exhibit brittle to pseudoplastic behaviour. Carbon-carbon is a unique all-carbon composite with carbon fibre embeded in carbon matrix and is known as an inverse composite. Due to their excellent thermo-structural properties, carbon-carbon composites are used in specialised application like re-entry nose-tips, leading edges, rocket nozzles, and aircraft brake discs apart from several industrial and biomedical applications. The multidirectional carbon-carbon product technology is versatile and offers design flexibility. This paper describes the multidirectional preform and carbon-carbon process technology and research and development activities within the country. Carbon-carbon product experience at DRDL has also been discussed. Development of carbon-carbon brake discs process technology using the liquid impregnation process is described. Further the test results on material characterisation, thermal, mechanical and tribological properties are presented.

  12. An experimental and DFT study of a disulfide-linked Schiff base: Synthesis, characterization and crystal structure of bis (3-methoxy-salicylidene-2-aminophenyl) disulfide in its anhydrous and monohydrate forms

    Science.gov (United States)

    Ferraresi-Curotto, Verónica; Echeverría, Gustavo A.; Piro, Oscar E.; Pis-Diez, Reinaldo; González-Baró, Ana C.

    2014-01-01

    A detailed structural and spectroscopic study of the disulfide Schiff base obtained from condensation of 2-aminothiophenol and o-vanillin is reported. It includes the analyses of the anhydrous and monohydrate forms of the title compound. Structures of both solids were resolved by X-ray diffraction methods. A comparison between experimental and theoretical results is presented. The conformational space was searched and geometries were optimized both in gas phase and including solvent effects. Vibrational (IR and Raman) and electronic spectra were measured and assigned with the help of computational methods based on the Density Functional Theory. Calculated MEP-derived atomic charges were calculated to predict coordination sites for metal complexes formation.

  13. Synthesis of thioglycolic acid by sodium disulfide%采用二硫化钠合成巯基乙酸

    Institute of Scientific and Technical Information of China (English)

    王晶; 王绍民

    2011-01-01

    介绍了采用二硫化钠合成巯基乙酸的新方法.以硫化钠和硫磺为基本原料制备二硫化钠,二硫化钠与氯乙酸钠合成2,2′-二硫代双乙酸钠,再用硫氢化钠和亚硫酸钠还原,经酸化、萃取、蒸馏,得到巯基乙酸.探讨了反应物浓度、反应温度、反应时间及酸化用酸的种类对巯基乙酸产品的影响.得出了最佳工艺条件:n硫化钠∶n硫∶n氯乙酸=1.4∶1.68∶1(物质的量比);硫化钠溶液的质量分数15%;合成2,2′-二硫代双乙酸钠时的反应温度45℃,反应时间30 min;采用盐酸酸化还原.在此条件下,产品收率可达92.6%.%A new method of synthesizing thioglycolic acid by sodium disulfide is presented. Sodium disulfide is prepared by sodium sulfide and sulfur, and 2, 2'-dithio sodium diacetate is prepared by sodium disulfide and chloroacetic acid. 2, 2'-Dithio sodium diacetate is reduced by sodium hydrosulfide and sodium sulfite, then acidated, extracted and distilled, the thioglycolic acid product is prepared finally. The effects of concentration of reactants, reaction temperature, reaction time and the type of acid on the product are discussed. The results show that the optimum conditions are as follows : the molar ratio of sodium sulfide, sulfur to chloroacetic acid is 1. 4 : 1. 68 : 1, the mass fraction of sodium sulfide is 15 % , the reaction temperature is 45 ℃, the reaction time is 30 minutes and hydrochloric acid is used as reactant. In these cases, the yield of thioglycolic acid can reach 92.6%.

  14. Formation of intersubunit disulfide bonds and properties of the single histidine and cysteine residues in each subunit relative to the decameric structure of cyanase.

    Science.gov (United States)

    Anderson, P M; Korte, J J; Holcomb, T A; Cho, Y G; Son, C M; Sung, Y C

    1994-05-27

    Reaction of the single cysteine residue in each subunit of cyanase with certain SH reagents gives an active decameric derivative that dissociates reversibly to an inactive dimer derivative (Anderson, P. M., Johnson, W. V., Korte, J. J., Xiong, X., Sung, Y.-c., and Fuchs, J. A. (1988) J. Biol. Chem. 263, 5674-5680). Reaction of mixed disulfide dimer derivatives of cyanase with dithiothreitol at 0 degree C results in formation of a disulfide bond between the subunits in the dimer. The disulfide dimer was inactive and did not associate to a decamer; the intersubunit disulfide bond could not be formed when the dimers were associated as a decamer. The two SH groups apparently are in close proximity to each other in the dissociated dimer but not when the dimer is associated to a decamer. Substitution of glycine for the cysteine residue or of tyrosine, asparagine, glycine, valine, or leucine for the single histidine residue in each subunit gave mutant enzymes that were active. However, H113N, H113Y, and C83G were unstable at low temperature and/or ionic strength, dissociating reversibly to an inactive dimer. Efficient reassociation required the presence of bicarbonate or cyanate analog. The results are consistent with a proposed single site per subunit model explaining apparent half-site binding of substrates and the requirement of decameric structure for activity.

  15. Stereoselective Michael Addition and Michael-aldol Tandem Reaction of Diorganyl Diselenides or Disulfides with Conjugated Alkynones Mediated by Samarium Diiodide

    Institute of Scientific and Technical Information of China (English)

    ZHENG Xing-Liang郑兴良; XU Xiao-Liang许孝良; ZHANG Yong-Min张永敏

    2004-01-01

    Stereoselective Michael addition and Michael-aldol tandem reaction of diorganyl diselenides and disulfides with conjugated alkynones mediated by samarium diiodide were studied. The reaction temperature was critical for the stereoselectivity. β-Organylselenoalkenones or β-organylthioalkenones and γ-organylselenoallylic alcohols orγ-organylthioallylic alcohols were prepared in good yields.

  16. Metal-free oxidative coupling of thiols to disulfides using guanidinium nitrate or nitro urea in the presence of silica sulfuric acid

    Indian Academy of Sciences (India)

    Arash Ghorbani-Choghamarani; Mohsen Nikoorazm; Hamid Goudarziafshar; Alireza Shokr; Hosein Almasi

    2011-07-01

    Efficient combination of nitro urea or guanidinium nitrate and silica sulfuric acid (SiO2OSO3H) as a new oxidizing system is able to oxidize a variety of aliphatic or aromatic thiols to the corresponding disulfides. The process reported here is operationally simple, environmentally benign and reactions have been mildly and heterogeneously performed in dichloromethane at room temperature.

  17. Insights into the mechanism of X-ray-induced disulfide-bond cleavage in lysozyme crystals based on EPR, optical absorption and X-ray diffraction studies

    International Nuclear Information System (INIS)

    Electron paramagnetic resonance (EPR) and online UV–visible absorption microspectrophotometry with X-ray crystallography have been used in a complementary manner to follow X-ray-induced disulfide-bond cleavage, to confirm a multi-track radiation-damage process and to develop a model of that process. Electron paramagnetic resonance (EPR) and online UV–visible absorption microspectrophotometry with X-ray crystallography have been used in a complementary manner to follow X-ray-induced disulfide-bond cleavage. Online UV–visible spectroscopy showed that upon X-irradiation, disulfide radicalization appeared to saturate at an absorbed dose of approximately 0.5–0.8 MGy, in contrast to the saturating dose of ∼0.2 MGy observed using EPR at much lower dose rates. The observations suggest that a multi-track model involving product formation owing to the interaction of two separate tracks is a valid model for radiation damage in protein crystals. The saturation levels are remarkably consistent given the widely different experimental parameters and the range of total absorbed doses studied. The results indicate that even at the lowest doses used for structural investigations disulfide bonds are already radicalized. Multi-track considerations offer the first step in a comprehensive model of radiation damage that could potentially lead to a combined computational and experimental approach to identifying when damage is likely to be present, to quantitate it and to provide the ability to recover the native unperturbed structure

  18. Evaluation of the combination of dimethyl disulfide and dazomet as an efficient methyl bromide alternative for cucumber production in China.

    Science.gov (United States)

    Mao, Liangang; Yan, Dongdong; Wang, Qiuxia; Li, Yuan; Ouyang, Canbin; Liu, Pengfei; Shen, Jin; Guo, Meixia; Cao, Aocheng

    2014-05-28

    The combination of dimethyl disulfide (DMDS) and dazomet (DZ) is a potential alternative to methyl bromide (MB) for soil disinfestation. The efficacy of DMDS plus DZ in controlling key soilborne pests was evaluated in a laboratory study and in two commercial cucumber greenhouses. Laboratory studies found that all of the combinations had positive synergistic effects on root-knot nematodes, two key soilborne fungi, and two major weed seeds. Greenhouse trials revealed that the combination of DMDS and DZ (30 + 25 g m(-2)) successfully suppressed Meloidogyne spp. root galling, sharply reduced the colony-forming units of Fusarium spp. and Phytophthora spp. on media, maintained high cucumber yields, and was not significantly different from MB or DMDS alone, but better than DZ alone. All of the chemical treatments provided significantly better results than the nontreated control. The results indicate that the combination of DMDS and DZ is an efficient MB alternative for cucumber production. PMID:24820184

  19. S-band Q-switched fiber laser using molybdenum disulfide (MoS2) saturable absorber

    Science.gov (United States)

    Ahmad, Harith; Afiq Ismail, Mohd; Suthaskumar, Muneswaran; Cheak Tiu, Zian; Wadi Harun, Sulaiman; Zamani Zulkifli, Mohd; Samikannu, Sathiyan; Sivaraj, Sivabalan

    2016-03-01

    In this letter, a molybdenum disulfide (MoS2) saturable absorber (SA) is fabricated using a simple drop cast method to generate Q-switched fiber laser operating in the S-band region (1460 nm-1530 nm). The MoS2 solution was prepared using the liquid phase exfoliation (LPE) method where MoS2 crystals were added into dimethylformamide (DMF) solvent and subsequently sonicated and centrifuged. They were then repeatedly dripped onto fiber ferrules and dried in an oven. The resultant Q-switched fiber laser starts with some physical disturbance when the pump power was set at 40 mW and continues to operate until the pump power reaches 120 mW. The resultant repetition rate varies with pump power between 27.17 to 101.17 kHz while the changes in pulse widths are from 3.0 to 1.4 μs.

  20. Q-switched waveguide laser based on two-dimensional semiconducting materials: tungsten disulfide and black phosphorous.

    Science.gov (United States)

    Tan, Yang; Guo, Zhinan; Ma, Linan; Zhang, Han; Akhmadaliev, Shavkat; Zhou, Shengqiang; Chen, Feng

    2016-02-01

    Owing to their unique properties, graphene-like two dimensional semiconducting materials, including Tungsten Disulfide (WS2) and Black Phosphorous (BP), have attracted increasing interest from basic research to practical applications. Herein, we demonstrated the ultrafast nonlinear saturable absorption response of WS2 and BP films in the waveguide structure. Through fabricating WS2 and BP films by evaporating the solutions on glass wafers. Saturable absorber films were attached onto the end-facet of the waveguide, which therefore constitutes a resonant cavity for the waveguide laser. Under a pump laser at 810 nm, we could obtain a stable Q-switched operation in the waveguide structure. This work indicated the significant potential of WS2 and BP for the ultrafast waveguide laser.

  1. Recent Advancement on the Optical Properties of Two-Dimensional Molybdenum Disulfide (MoS2 Thin Films

    Directory of Open Access Journals (Sweden)

    Mingxiao Ye

    2015-03-01

    Full Text Available The emergence of two-dimensional (2D materials has led to tremendous interest in the study of graphene and a series of mono- and few-layered transition metal dichalcogenides (TMDCs. Among these TMDCs, the study of molybdenum disulfide (MoS2 has gained increasing attention due to its promising optical, electronic, and optoelectronic properties. Of particular interest is the indirect to direct band-gap transition from bulk and few-layered structures to mono-layered MoS2, respectively. In this review, the study of these properties is summarized. The use of Raman and Photoluminescence (PL spectroscopy of MoS2 has become a reliable technique for differentiating the number of molecular layers in 2D MoS2.

  2. The role of thermal excitation in the tunneling-electron-induced reaction: Dissociation of dimethyl disulfide on Cu(111)

    Science.gov (United States)

    Motobayashi, Kenta; Kim, Yousoo; Ohara, Michiaki; Ueba, Hiromu; Kawai, Maki

    2016-01-01

    We found a thermally assisted increase in anharmonic coupling between the reaction coordinate and Csbnd H(D) stretch mode for the dissociation of a single dimethyl disulfide molecule on Cu(111) induced by inelastic tunneling electrons from a tip of scanning tunneling microscope (STM). The reaction order, i.e. the number of electrons required for a reaction, changes from two to one at elevated temperature while the Csbnd H(D) stretch mode is excited by tunneling electrons. The detailed reaction mechanism is studied through the quantitative analysis of the non-integer reaction order observed at intermediate temperature, where low energy vibrational mode originated from the hybridized state of molecule and substrate plays a key role.

  3. Quantifying changes in the cellular thiol-disulfide status during differentiation of B cells into antibody-secreting plasma cells

    DEFF Research Database (Denmark)

    Hansen, Rosa Rebecca Erritzøe; Otsu, Mieko; Braakman, Ineke;

    2013-01-01

    by the differentiation, steady-state levels of glutathionylated protein thiols are less than 0.3% of the total protein cysteines, even in fully differentiated cells, and the overall protein redox state is not affected until late in differentiation, when large-scale IgM production is ongoing. A general expansion......Plasma cells produce and secrete massive amounts of disulfide-containing antibodies. To accommodate this load on the secretory machinery, the differentiation of resting B cells into antibody-secreting plasma cells is accompanied by a preferential expansion of the secretory compartments of the cells...... of the ER does not affect global protein redox status until an extensive production of cargo proteins has started....

  4. Molecular dynamics simulations and 2D NOESY spectrum study on the different behaviors of glutathione disulfide in different solutions

    International Nuclear Information System (INIS)

    Highlights: ► Different behaviors of GSSG are observed in aqueous and DMSO solutions. ► The interesting phenomena have been proved by the MD simulations and 2D NMR experiment. ► The NOESY spectroscopy show good agreement with the MD simulations. - Abstract: All-atom molecular dynamics simulations and 2D nuclear Overhauser effect spectroscopy (NOESY) were used to study the interactions and conformations of glutathione disulfide (GSSG) in aqueous and DMSO solutions. GSSG showed variations in conformation in the two solutions, shifting between extended and folded states. Showing preference for extension in aqueous solution and folding for a long time in DMSO are observed by the simulations. The interesting phenomena have been proved by the 2D NMR experiment. The NMR experimental results show agreement with the molecular dynamics simulations.

  5. A trans-1,2 End-On Disulfide-Bridged Iron-Tetracarbene Dimer and Its Electronic Structure.

    Science.gov (United States)

    Meyer, Steffen; Krahe, Oliver; Kupper, Claudia; Klawitter, Iris; Demeshko, Serhiy; Bill, Eckhard; Neese, Frank; Meyer, Franc

    2015-10-19

    A disulfide-bridged diiron complex with [Fe-S-S-Fe] core, which represents an isomer of the common biological [2Fe-2S] ferredoxin-type clusters, was synthesized using strongly σ-donating macrocyclic tetracarbene capping ligands. Though the complex is quite labile in solution, single crystals were obtained, and the structure was elucidated by X-ray diffraction. The electron-rich iron-sulfur core is found to show rather unusual magnetic and electronic properties. Experimental data and density functional theory studies indicate extremely strong antiferromagnetic coupling (-J > 800 cm(-1)) between two low-spin iron(III) ions via the S2(2-) bridge, and the intense near-IR absorption characteristic for the [Fe-S-S-Fe] core was assigned to a S → Fe ligand-to-metal charge transfer transition.

  6. Q-switched waveguide laser based on two-dimensional semiconducting materials: tungsten disulfide and black phosphorous.

    Science.gov (United States)

    Tan, Yang; Guo, Zhinan; Ma, Linan; Zhang, Han; Akhmadaliev, Shavkat; Zhou, Shengqiang; Chen, Feng

    2016-02-01

    Owing to their unique properties, graphene-like two dimensional semiconducting materials, including Tungsten Disulfide (WS2) and Black Phosphorous (BP), have attracted increasing interest from basic research to practical applications. Herein, we demonstrated the ultrafast nonlinear saturable absorption response of WS2 and BP films in the waveguide structure. Through fabricating WS2 and BP films by evaporating the solutions on glass wafers. Saturable absorber films were attached onto the end-facet of the waveguide, which therefore constitutes a resonant cavity for the waveguide laser. Under a pump laser at 810 nm, we could obtain a stable Q-switched operation in the waveguide structure. This work indicated the significant potential of WS2 and BP for the ultrafast waveguide laser. PMID:26906854

  7. Hydrogen sulfide production during yeast fermentation causes the accumulation of ethanethiol, S-ethyl thioacetate and diethyl disulfide.

    Science.gov (United States)

    Kinzurik, Matias I; Herbst-Johnstone, Mandy; Gardner, Richard C; Fedrizzi, Bruno

    2016-10-15

    Hydrogen sulfide (H2S) is produced by yeast during winemaking and possesses off-flavors reminiscent of rotten eggs. The production of H2S during fermentation has also been associated in the finished wine with the rise of additional volatile sulfur compounds (VSCs) with strong aromas of cooked onions and vegetables. To characterize these more complex VSCs produced from H2S, we performed fermentations in synthetic grape juice. H2S production was manipulated experimentally by feeding increasing concentrations of sulfate to mutant strains that are unable to incorporate H2S efficiently as part of the sulfur assimilation pathway. In finished wines from these mutants, three VSCs - ethanethiol, S-ethyl thioacetate and diethyl disulfide - increased proportionally to H2S. (34)S-labeled sulfate fed to the MET17-deleted strain was incorporated into same three VSCs, demonstrating that they are formed directly from H2S. PMID:27173572

  8. Variation in the Subcellular Localization and Protein Folding Activity among Arabidopsis thaliana Homologs of Protein Disulfide Isomerase

    Directory of Open Access Journals (Sweden)

    Christen Y. L. Yuen

    2013-10-01

    Full Text Available Protein disulfide isomerases (PDIs catalyze the formation, breakage, and rearrangement of disulfide bonds to properly fold nascent polypeptides within the endoplasmic reticulum (ER. Classical animal and yeast PDIs possess two catalytic thioredoxin-like domains (a, a′ and two non-catalytic domains (b, b′, in the order a-b-b′-a′. The model plant, Arabidopsis thaliana, encodes 12 PDI-like proteins, six of which possess the classical PDI domain arrangement (AtPDI1 through AtPDI6. Three additional AtPDIs (AtPDI9, AtPDI10, AtPDI11 possess two thioredoxin domains, but without intervening b-b′ domains. C-terminal green fluorescent protein (GFP fusions to each of the nine dual-thioredoxin PDI homologs localized predominantly to the ER lumen when transiently expressed in protoplasts. Additionally, expression of AtPDI9:GFP-KDEL and AtPDI10: GFP-KDDL was associated with the formation of ER bodies. AtPDI9, AtPDI10, and AtPDI11 mediated the oxidative folding of alkaline phosphatase when heterologously expressed in the Escherichia coli protein folding mutant, dsbA−. However, only three classical AtPDIs (AtPDI2, AtPDI5, AtPDI6 functionally complemented dsbA−. Interestingly, chemical inducers of the ER unfolded protein response were previously shown to upregulate most of the AtPDIs that complemented dsbA−. The results indicate that Arabidopsis PDIs differ in their localization and protein folding activities to fulfill distinct molecular functions in the ER.

  9. A maurotoxin with constrained standard disulfide bridging: innovative strategy of chemical synthesis, pharmacology, and docking on K+ channels.

    Science.gov (United States)

    M'Barek, Sarrah; Lopez-Gonzalez, Ignacio; Andreotti, Nicolas; di Luccio, Eric; Visan, Violeta; Grissmer, Stephan; Judge, Susan; El Ayeb, Mohamed; Darbon, Hervé; Rochat, Hervé; Sampieri, François; Béraud, Evelyne; Fajloun, Ziad; De Waard, Michel; Sabatier, Jean-Marc

    2003-08-15

    Maurotoxin (MTX) is a 34-residue toxin that has been isolated initially from the venom of the scorpion Scorpio maurus palmatus. It presents a large number of pharmacological targets, including small conductance Ca2+-activated and voltage-gated K+ channels. Contrary to other toxins of the alpha-KTx6 family (Pi1, Pi4, Pi7, and HsTx1), MTX exhibits a unique disulfide bridge organization of the type C1-C5, C2-C6, C3-C4, and C7-C8 (instead of the conventional C1-C5, C2-C6, C3-C7, and C4-C8, herein referred to as Pi1-like) that does not prevent its folding along the classic alpha/beta scaffold of scorpion toxins. Here, we developed an innovative strategy of chemical peptide synthesis to produce an MTX variant (MTXPi1) with a conventional pattern of disulfide bridging without any alteration of the toxin chemical structure. This strategy was used solely to address the impact of half-cystine pairings on MTX structural properties and pharmacology. The data indicate that MTXPi1 displays some marked changes in affinities toward the target K+ channels. Computed docking analyses using molecular models of both MTXPi1 and the various voltage-gated K+ channel subtypes (Shaker B, Kv1.2, and Kv1.3) were found to correlate with MTXPi1 pharmacology. A functional map detailing the interaction between MTXPi1 and Shaker B channel was generated in line with docking experiments.

  10. Dual-degradable disulfide-containing PEI–Pluronic/DNA polyplexes: transfection efficiency and balancing protection and DNA release

    Directory of Open Access Journals (Sweden)

    Zhang L

    2013-09-01

    Full Text Available Lifen Zhang,* Zhenzhen Chen,* Yanfeng LiState Key Laboratory of Applied Organic Chemistry, Key Laboratory of Nonferrous Metals Chemistry and Resources Utilization of Gansu Province, College of Chemistry and Chemical Engineering, Institute of Biochemical Engineering and Environmental Technology, Lanzhou University, Lanzhou, People's Republic of China*These authors contributed equally to this workAbstract: Polymeric gene-delivery vectors to achieve lack of toxicity and a balance between protection and DNA release remains a formidable challenge. Incorporating intracellular environment-responsive degradable bonds is an appreciable step toward developing safer transfection agents. In this study, novel, dual-degradable polycation copolymers (Pluronic-diacrylate [PA]–polyethyleneimine [PEI]–SS were synthesized through the addition of low molecular weight (800 Da PEI cross-linked with SS (PEI-SS to PA. Three PA-PEI-SS copolymers (PA-PEI-SS1, 2, and 3 with different PEI-SS to Pluronic molar ratios were investigated and found to strongly condense plasmid DNA into positively charged nanoparticles with an average particle size of approximately 200 nm and to possess higher stability against DNase I digestion and sodium heparin. Disulfide and ester bonds of the copolymers were susceptible to intracellular redox conditions. In vitro experiments demonstrated that the PA-PEI-SS copolymers had significantly lower cytotoxicity and higher transfection efficiency in both BGC-823 and 293T cell lines than the controls of degradable PEI-SS and nondegradable 25 kDa PEI. Transfection activity was influenced by the PEI-SS content in the polymers and PA-PEI-SS1 showed the highest efficiency of the three copolymers. These studies suggest that these dual-degradable copolymers could be used as potential biocompatible gene delivery carriers.Keywords: Pluronic, PEI, gene vector, dual-degradable, disulfide-containing linker

  11. Preparation routes based on magnetron sputtering for tungsten disulfide (WS2) films for thin-film solar cells

    International Nuclear Information System (INIS)

    The semiconductor tungsten disulfide (WS2) exhibits van der Waals bonding, crystallizes in a layer-type structure and is of interest as an absorber layer for thin-film solar cells. In this review article different preparation routes for WS2 thin films, based on magnetron sputtering, are reviewed. Films prepared by direct magnetron sputtering, though exhibiting quite a good structural quality, are not or only poorly photoactive. This can be attributed to the generation of recombination centers, especially sulfur vacancies, during the ion bombardment of the films, due to the low defect-formation energy of tungsten disulfide, an intrinsic property of transition metal dichalcogenides. A promising preparation route, which leads to photoactive WS2 films, is a two-step process, where, in a first step, a sulfur-rich, X-ray amorphous tungsten sulfide is deposited at low substrate temperatures onto a thin metal film (Ni, Co). This film sandwich is after wards annealed in an ampoule in a sulfur atmosphere or in flowing gas with a sufficient H2S partial pressure. From in-situ transmission electron microscopy and energy-dispersive X-ray diffraction, it was found that the WS2 film crystallization with a pronounced (001) texture is closely related to the formation of the liquid (eutectic) metal-sulfur phase. Based on these in-situ investigations the growth of the 2-dimensional WS2 nanosheets from an amorphous WS3+x precursor can be described as an amorphous solid-liquid-crystalline solid process (SLS), somewhat similar to the well-known vapor-liquid-solid (VLS) process for the growth of whiskers or nanorods and nanotubes. Research opportunities, to overcome current limitations for a broad use of WS2 (and MoS2) as thin-film solar cell absorbers are given. (copyright 2008 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim) (orig.)

  12. Conformational instability governed by disulfide bonds partitions the dominant from subdominant helper T-cell responses specific for HIV-1 envelope glycoprotein gp120

    Science.gov (United States)

    Nguyen, Hong-Nam P.; Steede, N. Kalaya; Robinson, James E.; Landry, Samuel J.

    2015-01-01

    Most individuals infected with human immunodeficiency virus type 1 (HIV-1) generate a CD4+ T-cell response that is dominated by a few epitopes. Immunodominance may be counterproductive because a broad CD4+ T-cell response is associated with reduced viral load. Previous studies indicated that antigen three-dimensional structure controls antigen processing and presentation and therefore CD4+ T-cell epitope dominance. Dominant epitopes occur adjacent to the V1-V2, V3, and V4 loops because proteolytic antigen processing in the loops promotes presentation of adjacent sequences. In this study, three gp120 (strain JR-FL) variants were constructed, in which deletions of single outer-domain disulfide bonds were expected to introduce local conformational flexibility and promote presentation of additional CD4+ T-cell epitopes. Following mucosal immunization of C57BL/6 mice with wild-type or variant gp120 lacking the V3-flanking disulfide bond, the typical pattern of dominant epitopes was observed, suggesting that the disulfide bond posed no barrier to antigen presentation. In mice that lacked gamma interferoninducible lysosomal thioreductase (GILT), proliferative responses to the typically dominant epitopes of gp120 were selectively depressed, and the dominance pattern was rearranged. Deletion of the V3-flanking disulfide bond or one of the V4-flanking disulfide bonds partially restored highly proliferative responses to the typically dominant epitopes. These results reveal an acute dependence of dominant CD4+ T-cell responses on the native gp120 conformation. PMID:25944298

  13. Dithiocarbamates Strongly Inhibit Carbonic Anhydrases and Show Antiglaucoma Action in Vivo

    OpenAIRE

    Carta, Fabrizio; Aggarwal, Mayank; Maresca, Alfonso; Scozzafava, Andrea; McKenna, Robert; Masini, Emanuela; Supuran, Claudiu T.

    2012-01-01

    A series of dithiocarbamates was prepared by reaction of primary/secondary amines with carbon disulfide in the presence of bases. These compounds were tested for the inhibition of 4 human (h) isoforms of the zinc enzyme carbonic anhydrase, CA (EC 4.2.1.1), hCA I, II, IX and XII, involved in pathologies such as glaucoma (CA II and XII) or cancer (CA IX). Several low nanomolar inhibitors targeting these CAs were detected. X-ray crystal structure of hCA II adduct with morpholine dithiocarbamate ...

  14. Intracellular Delivery: Redox-Triggered Release of Moxifloxacin from Mesoporous Silica Nanoparticles Functionalized with Disulfide Snap-Tops Enhances Efficacy Against Pneumonic Tularemia in Mice (Small 27/2016).

    Science.gov (United States)

    Lee, Bai-Yu; Li, Zilu; Clemens, Daniel L; Dillon, Barbara Jane; Hwang, Angela A; Zink, Jeffrey I; Horwitz, Marcus A

    2016-07-01

    The drug trapping and intracellular release mechanism of redox-responsive disulfide snap-top mesoporous silica nanoparticles (MSN-SS-MXF) is depicted by J. I. Zink, M. A. Horwitz and co-workers on page 3690. Mesoporous silica nanoparticles with antibiotic (cyan) trapped within their pores by disulfide snap-tops are avidly ingested by macrophages. The intracellular redox potential reduces the disulfide (yellow) in the stalk (green/blue), releases the caps (orange) and frees drug to kill Francisella tularensis (green). Artwork by Bastian Ruehle. PMID:27412305

  15. Glutathione-mediated mesoporous carbon as a drug delivery nanocarrier with carbon dots as a cap and fluorescent tracer

    Science.gov (United States)

    Zhang, Yang; Han, Lu; Zhang, Yue; Chang, Yan-Qin; Chen, Xu-Wei; He, Rong-Huan; Shu, Yang; Wang, Jian-Hua

    2016-09-01

    This work describes a novel and general redox-responsive controlled drug delivery-release nanocarrier with mesoporous carbon nanoparticles (MCNs) gated by customized fluorescent carbon dots (CDs). The modification of MCNs with a disulfide unit enables the system to be sensitive to intracellular glutathione (GSH). The CDs anchoring onto the surface of the MCNs via an electrostatic interaction block the mesopores and thus prevent the leakage of doxorubicin (DOX) loaded inside the channel of the MCNs. Upon the addition of GSH at the physiological environment, the integrity of the system is disrupted due to the dissociation of the disulfide bond; meanwhile stripping the CDs opens the gate and thus triggers the rapid release of the encapsulated DOX. The fluorescence of the CDs is quenched/‘turned off’ when linking to the MCNs, while it is restored/‘turned on’ when detaching the CDs from the surface of the MCNs. Thus the fluorescent CDs serve as both a controllable drug release gatekeeper and a fluorescent probe for the visualization of the drug delivery process. By combining these inherent capabilities, the present drug delivery system may be a promising route for designing custom-made visual controlled-release nanodevices specifically governed by in situ stimulus in the cells.

  16. OX133, a monoclonal antibody recognizing protein-bound N-ethylmaleimide for the identification of reduced disulfide bonds in proteins.

    Science.gov (United States)

    Holbrook, Lisa-Marie; Kwong, Lai-Shan; Metcalfe, Clive L; Fenouillet, Emmanuel; Jones, Ian M; Barclay, A Neil

    2016-01-01

    In vivo, enzymatic reduction of some protein disulfide bonds, allosteric disulfide bonds, provides an important level of structural and functional regulation. The free cysteine residues generated can be labeled by maleimide reagents, including biotin derivatives, allowing the reduced protein to be detected or purified. During the screening of monoclonal antibodies for those specific for the reduced forms of proteins, we isolated OX133, a unique antibody that recognizes polypeptide resident, N-ethylmaleimide (NEM)-modified cysteine residues in a sequence-independent manner. OX133 offers an alternative to biotin-maleimide reagents for labeling reduced/alkylated antigens and capturing reduced/alkylated proteins with the advantage that NEM-modified proteins are more easily detected in mass spectrometry, and may be more easily recovered than is the case following capture with biotin based reagents. PMID:26986548

  17. Preparation of a disulfide-linked precipitative soluble support for solution-phase synthesis of trimeric oligodeoxyribonucleotide 3´-(2-chlorophenylphosphate building blocks

    Directory of Open Access Journals (Sweden)

    Amit M. Jabgunde

    2015-09-01

    Full Text Available The preparation of a disulfide-tethered precipitative soluble support and its use for solution-phase synthesis of trimeric oligodeoxyribonucleotide 3´-(2-chlorophenylphosphate building blocks is described. To obtain the building blocks, N-acyl protected 2´-deoxy-5´-O-(4,4´-dimethoxytritylribonucleosides were phosphorylated with bis(benzotriazol-1-yl 2-chlorophenyl phosphate. The “outdated” phosphotriester strategy, based on coupling of PV building blocks in conjunction with quantitative precipitation of the oligodeoxyribonucleotide with MeOH is applied. Subsequent release of the resulting phosphate and base-protected oligodeoxyribonucleotide trimer 3’-pTpdCBzpdGibu-5’ as its 3’-(2-chlorophenyl phosphate was achieved by reductive cleavage of the disulfide bond.

  18. Preparation of a disulfide-linked precipitative soluble support for solution-phase synthesis of trimeric oligodeoxyribonucleotide 3´-(2-chlorophenylphosphate) building blocks

    Science.gov (United States)

    Molina, Alejandro Gimenez; Virta, Pasi; Lönnberg, Harri

    2015-01-01

    Summary The preparation of a disulfide-tethered precipitative soluble support and its use for solution-phase synthesis of trimeric oligodeoxyribonucleotide 3´-(2-chlorophenylphosphate) building blocks is described. To obtain the building blocks, N-acyl protected 2´-deoxy-5´-O-(4,4´-dimethoxytrityl)ribonucleosides were phosphorylated with bis(benzotriazol-1-yl) 2-chlorophenyl phosphate. The “outdated” phosphotriester strategy, based on coupling of PV building blocks in conjunction with quantitative precipitation of the oligodeoxyribonucleotide with MeOH is applied. Subsequent release of the resulting phosphate and base-protected oligodeoxyribonucleotide trimer 3’-pTpdCBzpdGibu-5’ as its 3’-(2-chlorophenyl phosphate) was achieved by reductive cleavage of the disulfide bond. PMID:26664575

  19. Molybdenum Disulfide-Coated Lithium Vanadium Fluorophosphate Anode: Experiments and First-Principles Calculations.

    Science.gov (United States)

    Liu, Zhaomeng; Peng, Wenjie; Xu, Zhenming; Shih, Kaimin; Wang, Jiexi; Wang, Zhixing; Lv, Xiaojun; Chen, Jiangan; Li, Xinhai

    2016-08-23

    To develop a new anode material to meet the increasing demands of lithium-ion battery, MoS2 is used for the first time to modify the C/LiVPO4 F anode to improve its lithium-storage performance between 3 and 0.01 V. Morphological observations reveal that the MoS2 -modified C/LiVPO4 F particles (M-LVPF) are wrapped by an amorphous carbon as interlayer and layered MoS2 as external surface. Charge-discharge tests show that M-LVPF delivers a high reversible capacity of 308 mAh g(-1) at 50 mA g(-1) . After 300 cycles at 1.0 A g(-1) , a capacity retention of 98.7 % is observed. Moreover, it exhibits high rate capability with a specific capacity of 199 mAh g(-1) at 1.6 A g(-1) . Electrochemical impedance spectroscopy tests indicate that the lithium-ion diffusion and charge-exchange reaction at the surface of M-LVPF are greatly enhanced. First-principles calculations for the MoS2 (001)/C/LiVPO4 F (010) system demonstrate that the absorption of MoS2 on C/LiVPO4 F is exothermic and spontaneous and that the electron transfer at the MoS2 -absorbed C/LiVPO4 F surface is enhanced. PMID:27376792

  20. Formation of nanooctahedra in molybdenum disulfide and molybdenum diselenide using pulsed laser vaporization.

    Science.gov (United States)

    Parilla, Philip A; Dillon, Anne C; Parkinson, Bruce A; Jones, Kim M; Alleman, Jeff; Riker, Gerald; Ginley, David S; Heben, Michael J

    2004-05-20

    Pulsed laser vaporization has been used to produce nanooctahedra of MoS2 and MoSe2. The nanooctahedra primarily form in two- or three-layer nested octahedra, although nesting up to five layers has been observed. Tilting the TEM sample stage and mapping how the images of single particles transformed provided the evidence to verify their octahedral geometry. Analysis of 30 two- and three-layered octahedra showed that their outer edge lengths clustered at approximately 3.8 nm and approximately 5.1 nm, respectively. This discreet sizing and the high symmetry of these closed nanooctahedra represent the closest inorganic analogy yet to the carbon fullerenes. The geometrical implications for forming octahedra from these layered compounds are investigated by considering different atomic arrangements assuming either trigonal prismatic or octahedral coordination around the Mo atom and yields two possible configurations for the actual structure of the nanooctahedra. A preliminary survey of pulsed laser vaporization of other layered metal chalcogenides shows that these dichalcogenides differ in their tendency to form small closed layered fullerene-like structures. These materials can be ranked from highest tendency to lowest as follows: NbSe2, WS2, WSe2, SnS2, TaS2, GaS, ReS2, and MoTe2. PMID:18950101

  1. Accelerator-based production of the (99m)Tc-(186)Re diagnostic-therapeutic pair using metal disulfide targets (MoS2, WS2, OsS2).

    Science.gov (United States)

    Gott, Matthew D; Hayes, Connor R; Wycoff, Donald E; Balkin, Ethan R; Smith, Bennett E; Pauzauskie, Peter J; Fassbender, Michael E; Cutler, Cathy S; Ketring, Alan R; Wilbur, D Scott; Jurisson, Silvia S

    2016-08-01

    Novel, natural abundance metal disulfide targets were irradiated for 1h with a 10µA proton beam in a small, medical cyclotron. Osmium disulfide was synthesized by simple distillation and precipitation methods while MoS2 and WS2 were commercially available. The targets dissolved under mild conditions and were analyzed by γ-spectroscopy. Production rates and potential applications are discussed, including target recovery and recycling schemes for OsS2 and WS2. PMID:27236832

  2. Approach to Characterization of the Higher Order Structure of Disulfide-Containing Proteins Using Hydrogen/Deuterium Exchange and Top-Down Mass Spectrometry

    OpenAIRE

    Wang, Guanbo; Kaltashov, Igor A.

    2014-01-01

    Top-down hydrogen/deuterium exchange (HDX) with mass spectrometric (MS) detection has recently matured to become a potent biophysical tool capable of providing valuable information on higher order structure and conformational dynamics of proteins at an unprecedented level of structural detail. However, the scope of the proteins amenable to the analysis by top-down HDX MS still remains limited, with the protein size and the presence of disulfide bonds being the two most important limiting fact...

  3. The noncanonical disulfide bond as the important stabilizing element of the immunoglobulin fold of the Dr fimbrial DraE subunit.

    Science.gov (United States)

    Piatek, Rafał; Bruździak, Piotr; Wojciechowski, Marek; Zalewska-Piatek, Beata; Kur, Józef

    2010-02-23

    Fimbrial adhesins of pathogenic bacteria are linear protein associates responsible for binding to the specific host cell receptors. They are assembled via the chaperone/usher pathway conserved in Gram-negative bacteria. These adhesive organelles are characterized by the high resistance to dissociation and unfolding caused by temperature or chemical denaturants. The self-complemented (SC) recombinant subunits of adhesive structures make up the minimal model used to analyze stability phenomena of these organelles. The SC subunits are both highly stabilized thermodynamically and kinetically. They are characterized by a standard free energy of unfolding of 70-80 kJ/mol and a rate constant of unfolding of 10(-17) s(-1) (half-life of unfolding of 10(8) years at 25 degrees C). The DraE subunit of Dr fimbriae is characterized by a disulfide bond that joins the beginning of the A1 strand with the end of the B strand. Such localization is unique and differentiates this protein from other proteins of the Ig-like family. Sequence analysis shows that many protein subunits of adhesive structures possess cysteines that may form a potential disulfide bond homologous to that of DraE. In this paper, we investigate the influence of this noncanonical disulfide bond on the stability of DraE-sc by constructing a DraE-sc-DeltaSS mutant protein (Cys/Ala mutant). This construct unfolds thermally at a T(m) of 65.4 degrees C, more than 20 degrees C lower than that of the native DraE-sc protein, and possesses a different unfolding mechanism. The calculated standard free energy of unfolding of DraE-sc-DeltaSS is equal to 30 +/- 5 kJ/mol. This allows us to suggest that the disulfide bond is an important stabilizing feature of many fimbrial subunits.

  4. Rational Design of Disulfide Bonds Increases Thermostability of a Mesophilic 1,3-1,4-β-Glucanase from Bacillus terquilensis

    Science.gov (United States)

    Xu, Xin; Li, Qi

    2016-01-01

    1,3–1,4-β-glucanase is an important biocatalyst in brewing industry and animal feed industry, while its low thermostability often reduces its application performance. In this study, the thermostability of a mesophilic β-glucanase from Bacillus terquilensis was enhanced by rational design and engineering of disulfide bonds in the protein structure. Protein spatial configuration was analyzed to pre-exclude the residues pairs which negatively conflicted with the protein structure and ensure the contact of catalytic center. The changes in protein overall and local flexibility among the wild-type enzyme and the designated mutants were predicted to select the potential disulfide bonds for enhancement of thermostability. Two residue pairs (N31C-T187C and P102C-N125C) were chosen as engineering targets and both of them were proved to significantly enhance the protein thermostability. After combinational mutagenesis, the double mutant N31C-T187C/P102C-N125C showed a 48.3% increase in half-life value at 60°C and a 4.1°C rise in melting temperature (Tm) compared to wild-type enzyme. The catalytic property of N31C-T187C/P102C-N125C mutant was similar to that of wild-type enzyme. Interestingly, the optimal pH of double mutant was shifted from pH6.5 to pH6.0, which could also increase its industrial application. By comparison with mutants with single-Cys substitutions, the introduction of disulfide bonds and the induced new hydrogen bonds were proved to result in both local and overall rigidification and should be responsible for the improved thermostability. Therefore, the introduction of disulfide bonds for thermostability improvement could be rationally and highly-effectively designed by combination with spatial configuration analysis and molecular dynamics simulation. PMID:27100881

  5. Influence of liposome forms of the rhenium compounds and cis-platin on thiol-disulfide coefficient in the rats’ blood

    Directory of Open Access Journals (Sweden)

    I. V. Klenina

    2007-12-01

    Full Text Available Thiol-disulfide coefficient (TDC and its different modifications in model in vivo were studied. Introduction of the liposome forms of cluster rhenium compounds with organic ligands (CROL leads to both TDC increasing and to the constancy of the TDC. Thus, CROLs aren’t toxic agents and some compounds could mobilize organisms’ thiol defence system. Liposome form of cis-platin leads to the TDC decreasing. Important CROL capacities for its future medical treatment practice were shown.

  6. Bovine Insulin Filaments Induced by Reducing Disulfide Bonds Show a Different Morphology, Secondary Structure, and Cell Toxicity from Intact Insulin Amyloid Fibrils

    OpenAIRE

    Zako, Tamotsu; Sakono, Masafumi; Hashimoto, Naomi; Ihara, Masaki; Maeda, Mizuo

    2009-01-01

    Amyloid fibrils are associated with more than 20 diseases, including Alzheimer's disease and type II diabetes. Insulin is a 51-residue polypeptide hormone, with its two polypeptide chains linked by one intrachain and two interchain disulfide bonds, and has long been known to self-assemble in vitro into amyloid fibrils. We demonstrate here that bovine insulin forms flexible filaments in the presence of a reducing agent, Tris (2-carboxyethyl) phosphine. The insulin filaments, possibly formed du...

  7. Crystal Structures of the Reduced, Sulfenic Acid, and Mixed Disulfide Forms of SarZ, a Redox Active Global Regulator in Staphylococcus aureus

    Energy Technology Data Exchange (ETDEWEB)

    Poor, Catherine B.; Chen, Peng R.; Duguid, Erica; Rice, Phoebe A.; He, Chuan; (UC)

    2010-01-20

    SarZ is a global transcriptional regulator that uses a single cysteine residue, Cys{sup 13}, to sense peroxide stress and control metabolic switching and virulence in Staphylococcus aureus. SarZ belongs to the single-cysteine class of OhrR-MgrA proteins that play key roles in oxidative resistance and virulence regulation in various bacteria. We present the crystal structures of the reduced form, sulfenic acid form, and mixed disulfide form of SarZ. Both the sulfenic acid and mixed disulfide forms are structurally characterized for the first time for this class of proteins. The Cys{sup 13} sulfenic acid modification is stabilized through two hydrogen bonds with surrounding residues, and the overall DNA-binding conformation is retained. A further reaction of the Cys{sup 13} sulfenic acid with an external thiol leads to formation of a mixed disulfide bond, which results in an allosteric change in the DNA-binding domains, disrupting DNA binding. Thus, the crystal structures of SarZ in three different states provide molecular level pictures delineating the mechanism by which this class of redox active regulators undergoes activation. These structures help to understand redox-mediated virulence regulation in S. aureus and activation of the MarR family proteins in general.

  8. The thermodynamic stability of insulin disulfides is not affected by the C-domain of insulin-like growth factor 1

    Institute of Scientific and Technical Information of China (English)

    GUO; Zhanyun(郭占云); FENG; Youmin(冯佑民)

    2002-01-01

    Both Insulin and insulin-like growth factor 1 are members of insulin superfamily. They share homologous primary and tertiary structure as well as weakly overlapping biological activity. However, their folding behavior is different: insulin and its recombinant precursor (PIP) fold into one unique tertiary structure, while IGF-1 folds into two disulfides isomers with similar thermodynamic stability. To elucidate the molecular mechanism of their different folding behavior, we prepared a single-chain hybrid of insulin and IGF-1, [B10Glu]Ins/IGF-1(C), and studied its folding behavior compared with that of PIP and IGF-1. We also separated a major non-native disulfides isomer of the hybrid and studied its refolding. The data showed that the C-domain of IGF-1 did not affect the folding thermodynamics of insulin, that is, the primary structure of the hybrid encoded only one thermodynamically stable disulfides linkage. However, the folding kinetics of insulin was affected by the C-domain of IGF-1.

  9. Diallyl disulfide inhibits TNFα induced CCL2 release through MAPK/ERK and NF-Kappa-B signaling.

    Science.gov (United States)

    Bauer, D; Redmon, N; Mazzio, E; Taka, E; Reuben, J S; Day, A; Sadrud-Din, S; Flores-Rozas, H; Soliman, K F A; Darling-Reed, S

    2015-09-01

    TNFα receptors are constitutively overexpressed in tumor cells, correlating to sustain elevated NFκB and monocyte chemotactic protein-1 (MCP-1/CCL2) expression. The elevation of CCL2 evokes aggressive forms of malignant tumors marked by tumor associated macrophage (TAM) recruitment, cell proliferation, invasion and angiogenesis. Previously, we have shown that the organo-sulfur compound diallyl disulfide (DADS) found in garlic (Allium sativum) attenuates TNFα induced CCL2 production in MDA-MB-231 cells. In the current study, we explored the signaling pathways responsible for DADS suppressive effect on TNFα mediated CCL2 release using PCR Arrays, RT-PCR and western blots. The data in this study show that TNFα initiates a rise in NFκB mRNA, which is not reversed by DADS. However, TNFα induced heightened expression of IKKε and phosphorylated ERK. The expression of these proteins corresponds to increased CCL2 release that can be attenuated by DADS. CCL2 induction by TNFα was also lessened by inhibitors of p38 (SB202190) and MEK (U0126) but not JNK (SP 600125), all of which were suppressed by DADS. In conclusion, the obtained results indicate that DADS down regulates TNFα invoked CCL2 production primarily through reduction of IKKε and phosphorylated-ERK, thereby impairing MAPK/ERK, and NFκB pathway signaling. Future research will be required to evaluate the effects of DADS on the function and expression of TNFα surface receptors. PMID:26100848

  10. Three-dimensional Nitrogen-Doped Graphene Supported Molybdenum Disulfide Nanoparticles as an Advanced Catalyst for Hydrogen Evolution Reaction

    Science.gov (United States)

    Dong, Haifeng; Liu, Conghui; Ye, Haitao; Hu, Linping; Fugetsu, Bunshi; Dai, Wenhao; Cao, Yu; Qi, Xueqiang; Lu, Huiting; Zhang, Xueji

    2015-12-01

    An efficient three-dimensional (3D) hybrid material of nitrogen-doped graphene sheets (N-RGO) supporting molybdenum disulfide (MoS2) nanoparticles with high-performance electrocatalytic activity for hydrogen evolution reaction (HER) is fabricated by using a facile hydrothermal route. Comprehensive microscopic and spectroscopic characterizations confirm the resulting hybrid material possesses a 3D crumpled few-layered graphene network structure decorated with MoS2 nanoparticles. Electrochemical characterization analysis reveals that the resulting hybrid material exhibits efficient electrocatalytic activity toward HER under acidic conditions with a low onset potential of 112 mV and a small Tafel slope of 44 mV per decade. The enhanced mechanism of electrocatalytic activity has been investigated in detail by controlling the elemental composition, electrical conductance and surface morphology of the 3D hybrid as well as Density Functional Theory (DFT) calculations. This demonstrates that the abundance of exposed active sulfur edge sites in the MoS2 and nitrogen active functional moieties in N-RGO are synergistically responsible for the catalytic activity, whilst the distinguished and coherent interface in MoS2/N-RGO facilitates the electron transfer during electrocatalysis. Our study gives insights into the physical/chemical mechanism of enhanced HER performance in MoS2/N-RGO hybrids and illustrates how to design and construct a 3D hybrid to maximize the catalytic efficiency.

  11. Lignin-assisted exfoliation of molybdenum disulfide in aqueous media and its application in lithium ion batteries.

    Science.gov (United States)

    Liu, Wanshuang; Zhao, Chenyang; Zhou, Rui; Zhou, Dan; Liu, Zhaolin; Lu, Xuehong

    2015-06-01

    In this article, alkali lignin (AL)-assisted direct exfoliation of MoS2 mineral into single-layer and few-layer nanosheets in water is reported for the first time. Under optimized conditions, the concentration of MoS2 nanosheets in the obtained dispersion can be as high as 1.75 ± 0.08 mg mL(-1), which is much higher than the typical reported concentrations (MoS2 nanosheets are applied as electrode materials for lithium ion batteries, they show much improved electrochemical performance compared with the pristine MoS2 mineral because of the enhanced ion and electron transfer kinetics. This facile, scalable and eco-friendly aqueous-based process in combination with renewable and ultra-low-cost lignin opens up possibilities for large-scale fabrication of MoS2-based nanocomposites and devices. Moreover, herein we demonstrate that AL is also an excellent surfactant for exfoliation of many other types of layered materials, including graphene, tungsten disulfide and boron nitride, in water, providing rich opportunities for a wider range of applications. PMID:25970569

  12. Effects of γ-ray radiation on two-dimensional molybdenum disulfide (MoS{sub 2}) nanomechanical resonators

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jaesung; Feng, Philip X.-L., E-mail: philip.feng@case.edu [Department of Electrical Engineering and Computer Science, Case School of Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106 (United States); Krupcale, Matthew J. [Department of Electrical Engineering and Computer Science, Case School of Engineering, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106 (United States); Department of Physics, College of Arts and Sciences, Case Western Reserve University, 10900 Euclid Avenue, Cleveland, Ohio 44106 (United States)

    2016-01-11

    We report on experimental investigation and analysis of γ-ray radiation effects on two-dimensional molybdenum disulfide (MoS{sub 2}) drumhead nanomechanical resonators vibrating at megahertz frequencies. Given calibrated dosages of γ-ray radiation of ∼5000 photons with energy at 662 keV, upon exposure over 24 or 12 h, all the MoS{sub 2} resonators exhibit ∼0.5–2.1% resonance frequency upshifts due to the ionizing γ-ray induced charges and their interactions. The devices show γ-ray photon responsivity of ∼30–82 Hz/photon, with an intrinsic γ-ray sensitivity (limit of detection) estimated to approach ∼0.02–0.05 photon. After exposure expires, resonance frequencies return to an ordinary tendency where the frequency variations are dominated by long-term drift. These γ-ray radiation induced frequency shifts are distinctive from those due to pressure variation or surface adsorption mechanisms. The measurements and analyses show that MoS{sub 2} resonators are robust yet sensitive to very low dosage γ-ray, demonstrating a potential for ultrasensitive detection and early alarm of radiation in the very low dosage regime.

  13. Comparative elimination of dimethyl disulfide by maifanite and ceramic-packed biotrickling filters and their response to microbial community.

    Science.gov (United States)

    Chen, Xuequan; Liang, Zhishu; An, Taicheng; Li, Guiying

    2016-02-01

    Unpleasant odor emissions have traditionally occupied an important role in environmental concern. In this paper, twin biotrickling filters (BTFs) packed with different packing materials, seeded with Bacillus cereus GIGAN2, were successfully constructed to purify gaseous dimethyl disulfide (DMDS). The maifanite-packed BTF showed superior biodegradation capability to the ceramic-packed counterpart in terms of removal efficiency and elimination capacity under similar conditions. At an empty bed residence time of 123 s, 100% of DMDS could be removed by maifanite-packed BTF when DMDS inlet concentration was below 0.41 g m(-3). To achieve same effect, the inlet concentration must be lower than 0.25 g m(-3) for ceramic-packed BTF. The bacterial communities analyses found higher relative abundance of GIGAN2 in the maifanite-packed BTF, suggesting that maifanite is more suitable for GIGAN2 immobilization and for subsequent DMDS removal. This work indicates maifanite is a promising packing material for real odorous gases purification. PMID:26702514

  14. Non-disulfide-bridged peptides from Tityus serrulatus venom: Evidence for proline-free ACE-inhibitors.

    Science.gov (United States)

    Pucca, Manuela Berto; Cerni, Felipe Augusto; Pinheiro-Junior, Ernesto Lopes; Zoccal, Karina Furlani; Bordon, Karla de Castro Figueiredo; Amorim, Fernanda Gobbi; Peigneur, Steve; Vriens, Kim; Thevissen, Karin; Cammue, Bruno Philippe Angelo; Júnior, Ronaldo Bragança Martins; Arruda, Eurico; Faccioli, Lúcia Helena; Tytgat, Jan; Arantes, Eliane Candiani

    2016-08-01

    The present study purifies two T. serrulatus non-disulfide-bridged peptides (NDBPs), named venom peptides 7.2 (RLRSKG) and 8 (KIWRS) and details their synthesis and biological activity, comparing to the synthetic venom peptide 7.1 (RLRSKGKK), previously identified. The synthetic replicate peptides were subjected to a range of biological assays: hemolytic, antifungal, antiviral, electrophysiological, immunological and angiotensin-converting enzyme (ACE) inhibition activities. All venom peptides neither showed to be cytolytic nor demonstrated significant antifungal or antiviral activities. Interestingly, peptides were able to modulate macrophages' responses, increasing IL-6 production. The three venom peptides also demonstrated potential to inhibit ACE in the following order: 7.2>7.1>8. The ACE inhibition activity was unexpected, since peptides that display this function are usually proline-rich peptides. In attempt to understand the origin of such small peptides, we discovered that the isolated peptides 7.2 and 8 are fragments of the same molecule, named Pape peptide precursor. Furthermore, the study discusses that Pape fragments could be originated from a post-splitting mechanism resulting from metalloserrulases and other proteinases cleavage, which can be seen as a clever mechanism used by the scorpion to enlarge its repertoire of venom components. Scorpion venom remains as an interesting source of bioactive proteins and this study advances our knowledge about three NDBPs and their biological activities. PMID:27221550

  15. The Potato Sucrose Transporter StSUT1 Interacts with a DRM-Associated Protein Disulfide Isomerase

    Institute of Scientific and Technical Information of China (English)

    Undine Krügel; Hong-Xia He; Konstanze Gier; Jana Reins; Izabela Chincinska; Bernhard Grimm; Waltraud X. Schulze; Christina Kühn

    2012-01-01

    Organization of proteins into complexes is crucial for many cellular functions.Recently,the SUT1 protein was shown to form homodimeric complexes,to be associated with lipid raft-like microdomains in yeast as well as in plants and to undergo endocytosis in response to brefeldin A.We therefore aimed to identify SUT1-interacting proteins that might be involved in dimerization,endocytosis,or targeting of SUT1 to raft-like microdomains.Therefore,we identified potato membrane proteins,which are associated with the detergent-resistant membrane (DRM) fraction.Among the proteins identified,we clearly confirmed StSUT1 as part of DRM in potato source leaves.We used the yeast two-hybrid split ubiquitin system (SUS) to systematically screen for interaction between the sucrose transporter StSUT1 and other membraneassociated or soluble proteins in vivo.The SUS screen was followed by immunoprecipitation using affinity-purified StSUT1-specific peptide antibodies and mass spectrometric analysis of co-precipitated proteins.A large overlap was observed between the StSUT1-interacting proteins identified in the co-immunoprecipitation and the detergent-resistant membrane fraction.One of the SUT1-interacting proteins,a protein disulfide isomerase (PDI),interacts also with other sucrose transporter proteins.A potential role of the PDI as escort protein is discussed.

  16. Cysteine Specific Targeting of the Functionally Distinct Peroxiredoxin and Glutaredoxin Proteins by the Investigational Disulfide BNP7787

    Directory of Open Access Journals (Sweden)

    Aulma R. Parker

    2015-03-01

    Full Text Available Glutaredoxin (Grx, peroxiredoxin (Prx, and thioredoxin (Trx are redoxin family proteins that catalyze different types of chemical reactions that impact cell growth and survival through functionally distinct intracellular pathways. Much research is focused on understanding the roles of these redoxin proteins in the development and/or progression of human diseases. Grx and Prx are overexpressed in human cancers, including human lung cancers. BNP7787 is a novel investigational agent that has been evaluated in previous clinical studies, including non-small cell lung cancer (NSCLC studies. Herein, data from activity assays, mass spectrometry analyses, and X-ray crystallographic studies indicate that BNP7787 forms mixed disulfides with select cysteine residues on Grx and Prx and modulates their function. Studies of interactions between BNP7787 and Trx have been conducted and reported separately. Despite the fact that Trx, Grx, and Prx are functionally distinct proteins that impact oxidative stress, cell proliferation and disease processes through different intracellular pathways, BNP7787 can modify each protein and appears to modulate function through mechanisms that are unique to each target protein. Tumor cells are often genomically heterogeneous containing subpopulations of cancer cells that often express different tumor-promoting proteins or that have multiple dysregulated signaling pathways modulating cell proliferation and drug resistance. A multi-targeted agent that simultaneously modulates activity of proteins important in mediating cell proliferation by functionally distinct intracellular pathways could have many potentially useful therapeutic applications.

  17. New ruthenium(II) carbonyl complexes bearing disulfide Schiff base ligands and their applications as catalyst for some organic transformations

    Science.gov (United States)

    Prakash, Govindan; Viswanathamurthi, Periasamy

    2014-08-01

    Schiff base disulfide ligands (H2L1-6) were synthesized from the condensation of cystamine with salicylaldehyde(H2L1), 5-chlorosalicylaldehyde(H2L2), o-vanillin(H2L3), 2-hydroxyacetophenone(H2L4), 3-methyl-2-hydroxyacetophenone(H2L5), and 2-hydroxy-1-naphthaldehyde(H2L6). H2L1-6 reacts with the ruthenium precursor complex [RuHCl(CO)(PPh3)3] in benzene giving rise to six new ruthenium(II) complexes of general formula [Ru(CO)L1-6]. Characterization of the new complexes was carried out by using elemental and spectral (IR, UV-Vis, NMR (1H and 13C) and Mass) techniques. An octahedral geometry was assigned for all the complexes based on the spectral data obtained. The catalytic efficiency of the new complexes in aldehyde to amide conversion in the presence of NaHCO3, N-alkylation of aniline in the presence of t-BuOK, and transfer hydrogenation of ketones in the presence of iPrOH/KOH reactions were studied. Furthermore, the effect of solvents and catalyst/substrate ratio on the catalytic aldehyde to amide conversion were also discussed.

  18. Highly efficient hydrogen evolution reaction using crystalline layered three-dimensional molybdenum disulfides grown on graphene film.

    Energy Technology Data Exchange (ETDEWEB)

    Behranginia, Amirhossein; Asadi, Mohammad; Liu, Cong; Yasaei, Poya; Kumar, Bijandra; Phillips, Patrick; Foroozan, Tara; Waranius, Joseph C.; Kim, Kibum; Abiade, Jeremiah; Klie, Robert F.; Curtiss, Larry A.; Salehi-Khojin, Amin

    2016-01-26

    Electrochemistry is central to applications in the field of energy storage and generation. However, it has advanced far more slowly over the last two decades, mainly because of a lack of suitable and affordable catalysts. Here, we report the synthesis of highly crystalline layered three-dimensional (3D) molybdenum disulfide (MoS2) catalysts with bare Mo-edge atoms and demonstrate their remarkable performance for the hydrogen evolution reaction (HER). We found that Mo-edge-terminated 3D MoS2 directly grown on graphene film exhibits a remarkable exchange current density (18.2 mu A cm(-2)) and turnover frequency (>4 S-1) for HER. The obtained exchange current density is 15.2 and 2.3 times higher than that of MoS2/graphene and MoS2/Au catalysts, respectively, both with sulfided Mo-edge atoms. An easily scalable and robust growth process on a wide variety of substrates, along with prolonged stability, suggests that this material is a promising catalyst in energy-related applications.

  19. Band Gap Engineering and Layer-by-Layer Band Gap Mapping of Selenium-doped Molybdenum Disulfide

    Energy Technology Data Exchange (ETDEWEB)

    Gong, Yongji [Rice University; Liu, Zheng [Rice University; Lupini, Andrew R [ORNL; Lin, Junhao [ORNL; Pantelides, Sokrates T [ORNL; Pennycook, Stephen J [ORNL; Zhou, Wu [ORNL; Ajayan, Pullikel M [Rice University

    2014-01-01

    Ternary two-dimensional dichalcogenide alloys exhibit compositionally modulated electronic structure and hence, control of dopant concentration within each layer of these layered compounds provides a powerful way to modify their properties. The challenge then becomes quantifying and locating the dopant atoms within each layer in order to better understand and fine-tune the desired properties. Here we report the synthesis of selenium substitutionally doped molybdenum disulfide atomic layers, with a broad range of selenium concentrations, resulting in band gap modulations of over 0.2 eV. Atomic scale chemical analysis using Z-contrast imaging provides direct maps of the dopant atom distribution in individual MoS2 layers and hence a measure of the local band gaps. Furthermore, in a bilayer structure, the dopant distribution of each layer is imaged independently. We demonstrate that each layer in the bilayer contains similar doping levels, randomly distributed, providing new insights into the growth mechanism and alloying behavior in two-dimensional dichalcogenide atomic layers. The results show that growth of uniform, ternary, two-dimensional dichalcogenide alloy films with tunable electronic properties is feasible.

  20. The disulfide compound α-lipoic acid and its derivatives: A novel class of anticancer agents targeting mitochondria.

    Science.gov (United States)

    Dörsam, Bastian; Fahrer, Jörg

    2016-02-01

    The endogenous disulfide α-lipoic acid (LA) is an essential mitochondrial co-factor. In addition, LA and its reduced counterpart dihydro lipoic acid form a potent redox couple with antioxidative functions, for which it is used as dietary supplement and therapeutic. Recently, it has gained attention due to its cytotoxic effects in cancer cells, which is the key aspect of this review. We initially recapitulate the dietary occurrence, gastrointestinal absorption and pharmacokinetics of LA, illustrating its diverse antioxidative mechanisms. We then focus on its mode of action in cancer cells, in which it triggers primarily the mitochondrial pathway of apoptosis, whereas non-transformed primary cells are hardly affected. Furthermore, LA impairs oncogenic signaling and displays anti-metastatic potential. Novel LA derivatives such as CPI-613, which target mitochondrial energy metabolism, are described and recent pre-clinical studies are presented, which demonstrate that LA and its derivatives exert antitumor activity in vivo. Finally, we highlight clinical studies currently performed with the LA analog CPI-613. In summary, LA and its derivatives are promising candidates to complement the arsenal of established anticancer drugs due to their mitochondria-targeted mode of action and non-genotoxic properties. PMID:26604131

  1. Elemental Sulfur and Molybdenum Disulfide Composites for Li-S Batteries with Long Cycle Life and High-Rate Capability.

    Science.gov (United States)

    Dirlam, Philip T; Park, Jungjin; Simmonds, Adam G; Domanik, Kenneth; Arrington, Clay B; Schaefer, Jennifer L; Oleshko, Vladimir P; Kleine, Tristan S; Char, Kookheon; Glass, Richard S; Soles, Christopher L; Kim, Chunjoong; Pinna, Nicola; Sung, Yung-Eun; Pyun, Jeffrey

    2016-06-01

    The practical implementation of Li-S technology has been hindered by short cycle life and poor rate capability owing to deleterious effects resulting from the varied solubilities of different Li polysulfide redox products. Here, we report the preparation and utilization of composites with a sulfur-rich matrix and molybdenum disulfide (MoS2) particulate inclusions as Li-S cathode materials with the capability to mitigate the dissolution of the Li polysulfide redox products via the MoS2 inclusions acting as "polysulfide anchors". In situ composite formation was completed via a facile, one-pot method with commercially available starting materials. The composites were afforded by first dispersing MoS2 directly in liquid elemental sulfur (S8) with sequential polymerization of the sulfur phase via thermal ring opening polymerization or copolymerization via inverse vulcanization. For the practical utility of this system to be highlighted, it was demonstrated that the composite formation methodology was amenable to larger scale processes with composites easily prepared in 100 g batches. Cathodes fabricated with the high sulfur content composites as the active material afforded Li-S cells that exhibited extended cycle lifetimes of up to 1000 cycles with low capacity decay (0.07% per cycle) and demonstrated exceptional rate capability with the delivery of reversible capacity up to 500 mAh/g at 5 C. PMID:27171646

  2. Effect of Polymer Addition on the Structure and Hydrogen Evolution Reaction Property of Nanoflower-Like Molybdenum Disulfide

    Directory of Open Access Journals (Sweden)

    Xianwen Zeng

    2015-10-01

    Full Text Available Nano-structured molybdenum disulfide (MoS2 catalysts have been extensively developed for the hydrogen evolution reaction (HER. Herein, a novel hydrothermal intercalation approach is employed to fabricate nanoflower-like 2H–MoS2 with the incorporation of three polymers, polyvinylpyrrolidone (PVP, polyvinyl alcohol (PVA, and polyethylenimine (PEI. The as-prepared MoS2 specimens were characterized by techniques of scanning electron microscope (SEM, transmission electron microscopy (TEM, X-ray diffraction (XRD, together with Raman and Fourier transform infrared spectroscopy (FTIR. The HER properties of these lamellar nanoflower-like composites were evaluated using electrochemical tests of linear sweep voltammetry (LSV and electrochemical impedance spectroscopy (EIS. The existent polymer enlarges the interlayer spacing of the lamellar MoS2, and reduces its stacked thickness. The lamellar MoS2 samples exhibit a promoting activity in HER at low additions of these three polymers (0.04 g/g MoS2 for PVA and PEI, and 0.08 g/g MoS2 for PVP. This can be attributed to the fact that the expanded interlayer of MoS2 can offer abundant exposed active sites for HER. Conversely, high additions of the polymers exert an obvious interference in the HER activity of the lamellar MoS2. Compared with the samples of MoS2/PVP–0.08 and MoS2/PEI–0.04, the MoS2/PVA–0.04 composite exhibits excellent activity in HER, in terms of higher current density and lower onset potential.

  3. Secreted APE1/Ref-1 inhibits TNF-α-stimulated endothelial inflammation via thiol-disulfide exchange in TNF receptor.

    Science.gov (United States)

    Park, Myoung Soo; Choi, Sunga; Lee, Yu Ran; Joo, Hee Kyoung; Kang, Gun; Kim, Cuk-Seong; Kim, Soo Jin; Lee, Sang Do; Jeon, Byeong Hwa

    2016-03-11

    Apurinic apyrimidinic endonuclease 1/Redox factor-1 (APE1/Ref-1) is a multifunctional protein with redox activity and is proved to be secreted from stimulated cells. The aim of this study was to evaluate the functions of extracellular APE1/Ref-1 with respect to leading anti-inflammatory signaling in TNF-α-stimulated endothelial cells in response to acetylation. Treatment of TNF-α-stimulated endothelial cells with an inhibitor of deacetylase that causes intracellular acetylation, considerably suppressed vascular cell adhesion molecule-1 (VCAM-1). During TSA-mediated acetylation in culture, a time-dependent increase in secreted APE1/Ref-1 was confirmed. The acetyl moiety of acetylated-APE1/Ref-1 was rapidly removed based on the removal kinetics. Additionally, recombinant human (rh) APE1/Ref-1 with reducing activity induced a conformational change in rh TNF-α receptor 1 (TNFR1) by thiol-disulfide exchange. Following treatment with the neutralizing anti-APE1/Ref-1 antibody, inflammatory signals via the binding of TNF-α to TNFR1 were remarkably recovered, leading to up-regulation of reactive oxygen species generation and VCAM-1, in accordance with the activation of p66(shc) and p38 MAPK. These results strongly indicate that anti-inflammatory effects in TNF-α-stimulated endothelial cells by acetylation are tightly linked to secreted APE1/Ref-1, which inhibits TNF-α binding to TNFR1 by reductive conformational change, with suggestion as an endogenous inhibitor of vascular inflammation.

  4. Calcium-activated potassium channels in insect pacemaker neurons as unexpected target site for the novel fumigant dimethyl disulfide.

    Science.gov (United States)

    Gautier, Hélène; Auger, Jacques; Legros, Christian; Lapied, Bruno

    2008-01-01

    Dimethyl disulfide (DMDS), a plant-derived insecticide, is a promising fumigant as a substitute for methyl bromide. To further understand the mode of action of DMDS, we examined its effect on cockroach octopaminergic neurosecretory cells, called dorsal unpaired median (DUM) neurons, using whole-cell patch-clamp technique, calcium imaging and antisense oligonucleotide strategy. At low concentration (1 microM), DMDS modified spontaneous regular spike discharge into clear bursting activity associated with a decrease of the amplitude of the afterhyperpolarization. This effect led us to suspect alterations of calcium-activated potassium currents (IKCa) and [Ca(2+)](i) changes. We showed that DMDS reduced amplitudes of both peak transient and sustained components of the total potassium current. IKCa was confirmed as a target of DMDS by using iberiotoxin, cadmium chloride, and pSlo antisense oligonucleotide. In addition, we showed that DMDS induced [Ca(2+)](i) rise in Fura-2-loaded DUM neurons. Using calcium-free solution, and (R,S)-(3,4-dihydro-6,7-dimethoxy-isoquinoline-1-yl)-2-phenyl-N,N-di-[2-(2,3,4-trimethoxy-phenyl)ethyl]-acetamide (LOE 908) [an inhibitor of transient receptor potential (TRP)gamma], we demonstrated that TRPgamma initiated calcium influx. By contrast, omega-conotoxin GVIA (an inhibitor of N-type high-voltage-activated calcium channels), did not affect the DMDS-induced [Ca(2+)](i) rise. Finally, the participation of the calcium-induced calcium release mechanism was investigated using thapsigargin, caffeine, and ryanodine. Our study revealed that DMDS-induced elevation in [Ca(2+)](i) modulated IKCa in an unexpected bell-shaped manner via intracellular calcium. In conclusion, DMDS affects multiple targets, which could be an effective way to improve pest control efficacy of fumigation. PMID:17942746

  5. Disulfide-crosslinked nanomicelles confer cancer-specific drug delivery and improve efficacy of paclitaxel in bladder cancer

    Science.gov (United States)

    Pan, Amy; Zhang, Hongyong; Li, Yuanpei; Lin, Tzu-yin; Wang, Fuli; Lee, Joyce; Cheng, Mingshan; Dall'Era, Marc; Li, Tianhong; deVere White, Ralph; Pan, Chong-Xian; Lam, Kit S.

    2016-10-01

    Chemotherapy commonly used in the treatment of advanced bladder cancer is only moderately effective and associated with significant toxicity. There has been no appreciable improvement in overall survival over the last three decades. The goal of this project is to develop and characterize bladder cancer-specific nanometer-scale micelles loaded with the chemotherapeutic drug paclitaxel (PTX) and determine the anti-tumor activity and toxicity. Micelle-building-material telodendrimers were synthesized through the stepwise conjugation of eight cholic acid units at one terminus of polyethylene glycol (PEG) and a bladder cancer-specific targeting peptide named PLZ4 at the other terminus. To synthesize disulfide-crosslinked PLZ4 nanomicelles (DC-PNM), cysteine was introduced between the cholic acid and PEG. DC-PNM-PTX was synthesized through the evaporation method by loading PTX in the core. The loading capacity of PTX in DC-PNM was 25% (W/W). The loading efficiency was over 99%. DC-PNM-PTX was spherical with the median size of 25 nm. The stability of DC-PNM-PTX was determined in a solution containing sodium docecyl sulfate (SDS). It was stable in a SDS solution, but dissolved within 5 min after the addition of glutathione at the physiological intracellular concentration of 10 mM. In vivo targeting and anti-tumor activity were determined in immunodeficient mice carrying patient-derived bladder cancer xenografts (PDXs). After intravenous administration, DC-PNM specifically targeted the bladder cancer PDXs, but very little to the lung cancer xenografts in the same mice (p cancer xenografts in vivo, and improved the anti-cancer efficacy of PTX.

  6. Inhibitory effects of diallyl disulfide on the production of inflammatory mediators and cytokines in lipopolysaccharide-activated BV2 microglia

    Energy Technology Data Exchange (ETDEWEB)

    Park, Hye Young [Department of Biochemistry, Dongeui University College of Oriental Medicine, Busan 614-714 (Korea, Republic of); Department of Pharmacy, Pusan National University, Busan 609-735 (Korea, Republic of); Kim, Nam Deuk [Department of Pharmacy, Pusan National University, Busan 609-735 (Korea, Republic of); Kim, Gi-Young [Department of Marine Life Sciences, Jeju National University, Jeju 690-756 (Korea, Republic of); Hwang, Hye Jin [Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan 614-714 (Korea, Republic of); Department of Food and Nutrition, College of Human Ecology, Dongeui University, Busan 614-714 (Korea, Republic of); Kim, Byung-Woo [Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan 614-714 (Korea, Republic of); Department of Life Science and Biotechnology, College of Natural Science, Dongeui University, Busan 614-714 (Korea, Republic of); Department of Biomaterial Control, Graduate School, Dongeui University, Busan 614-714 (Korea, Republic of); Kim, Wun Jae [Department of Urology, College of Medicine, Chungbuk National University, Cheongju, Chungbuk 361-763 (Korea, Republic of); Choi, Yung Hyun, E-mail: choiyh@deu.ac.kr [Department of Biochemistry, Dongeui University College of Oriental Medicine, Busan 614-714 (Korea, Republic of); Anti-Aging Research Center and Blue-Bio Industry RIC, Dongeui University, Busan 614-714 (Korea, Republic of); Department of Biomaterial Control, Graduate School, Dongeui University, Busan 614-714 (Korea, Republic of)

    2012-07-15

    Diallyl disulfide (DADS), a main organosulfur component responsible for the diverse biological effects of garlic, displays a wide variety of internal biological activities. However, the cellular and molecular mechanisms underlying DADS' anti-inflammatory activity remain poorly understood. In this study, therefore, the anti-inflammatory effects of DADS were studied to investigate its potential therapeutic effects in lipopolysaccharide (LPS)-stimulated BV2 microglia. We found that pretreatment with DADS prior to treatment with LPS significantly inhibited excessive production of nitric oxide (NO) and prostaglandin E{sub 2} (PGE{sub 2}) in a dose-dependent manner. The inhibition was associated with down-regulation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression. DADS also attenuated the production of pro-inflammatory cytokines and chemokines, including interleukin-1β (IL-1β), tumor necrosis factor (TNF)-α, and monocyte chemoattractant protein-1 (MCP-1) by suppressing the expression of mRNAs for these proteins. The mechanism underlying this protective effect might be related to the inhibition of nuclear factor-kappaB, Akt and mitogen-activated protein kinase signaling pathway activation in LPS-stimulated microglial cells. These findings indicated that DADS is potentially a novel therapeutic candidate for the treatment of various neurodegenerative diseases. -- Highlights: ► DADS attenuates production of NO and PGE2 in LPS-activated BV2 microglia. ► DADS downregulates levels of iNOS and COX-2. ► DADS inhibits production and expression of inflammatory cytokines and chemokine. ► DADS exhibits these effects by suppression of NF-κB, PI3K/Akt and MAPKs pathways.

  7. Purification and characterization of a stable cysteine protease ervatamin B, with two disulfide bridges, from the latex of Ervatamia coronaria.

    Science.gov (United States)

    Kundu, S; Sundd, M; Jagannadham, M V

    2000-02-01

    Latex of the medicinal plant Ervatamia coronaria was found to contain at least three cysteine proteases with high proteolytic activity, called ervatamins. One of these proteases, named ervatamin B, has been purified to homogeneity using ion-exchange chromatography and crystallization. The molecular mass of the enzyme was estimated to be 26 000 Da by SDS-PAGE and gel filtration. The extinction coefficient (epsilon(1%)(280 nm)) of the enzyme was 20.5 with 7 tryptophan and 10 tyrosine residues per molecule. The enzyme hydrolyzed denatured natural substrates such as casein, azoalbumin, and azocasein with a high specific activity. In addition, it showed amidolytic activity toward N-succinyl-alanine-alanine-alanine-p-nitroanilide with an apparent K(m) and K(cat) of 6.6 +/- 0.5 mM and 1.87 x 10(2) s(-)(1), respectively. The pH optima was 6.0-6.5 with azocasein as substrate and 7.0-7.5 with azoalbumin as substrate. The temperature optimum was around 50-55 degrees C. The enzyme was basic with an isoelectric point of 9.35 and had no carbohydrate content. Both the proteolytic and amidolytic activity of the enzyme was strongly inhibited by thiol-specific inhibitors. Interestingly, the enzyme had only two disulfide bridges versus three as in most plant cysteine proteases of the papain superfamily. The enzyme was relatively stable toward pH, denaturants, temperature, and organic solvents. Polyclonal antibodies raised against the pure enzyme gave a single precipitin line in Ouchterlony's double immunodiffusion and typical color in ELISA. Other related proteases do not cross-react with the antisera to ervatamin B showing that the enzyme is immunologically distinct. The N-terminal sequence showed conserved amino acid residues and considerable similarity to typical plant cysteine proteases. PMID:10691612

  8. Dietary diallyl disulfide supplementation attenuates ethanol-mediated pulmonary vitamin D speciate depletion in C57Bl/6 mice

    Science.gov (United States)

    McCaskill, Michael L.; Hottor, Henry T.; Sapkota, Muna; Wyatt, Todd A.

    2016-01-01

    Background Slightly more than 5 % of the United States population heavily consumes ethanol, i.e., more than 14 drinks for men and 7 drinks for women a week. Chronic ethanol consumption can result in increased liver disease, reduced recovery from burn injury, and more frequent and severe respiratory infections. Chronic ethanol over-consumption also leads to vitamin D dysmetabolism and depletion. Vitamin D is a fat-soluble pro-hormone that regulates musculoskeletal health, cellular proliferation/differentiation, and innate and adaptive immune response. Methods In this study, C57BL/6 mice were fed 20 % ethanol in their water ad libitum for 7 weeks. Some mice were fed either a standard chow or a modified diet containing 0.15 μg/day of diallyl disulfide (DADS). Whole blood, lung tissue, and bronchial alveolar lavage fluid (BALF) were collected at sacrifice and analyzed for 25(OH) D3, 1,25 (OH)2D3, vitamin D receptor VDR, CYP2E1, and CYP27B1 levels. Results Ethanol reduced 25(OH) D3 and 1,25 (OH)2D3 in lung tissue and BALF on average 31 %. The largest ethanol-mediated reduction was in the 1,25 (OH)2D3 (42 %) measured in the BALF. Dietary supplementation of DADS restored BALF and lung tissue protein of 25(OH) D3 and 1,25(OH)2D3 to control levels. Chronic ethanol consumption also resulted in tissue increases of vitamin D response (VDR) protein, Cyp2E1, and reductions in vitamin D-activating enzyme CYP27B1. All three of these effects were attenuated by dietary supplementation of DADS. Conclusions In conclusion, the pulmonary metabolic disturbances mediated by chronic ethanol consumption as measured by 1,25(OH)2D3 protein levels, epithelial lining fluid, and lung tissue can be ameliorated by dietary supplementation of DADS in C57BL/6 mice.

  9. In vitro controlled release of cisplatin from gold-carbon nanobottles via cleavable linkages

    Directory of Open Access Journals (Sweden)

    Li J

    2015-12-01

    Full Text Available Jian Li,1 Sia Lee Yoong,2 Wei Jiang Goh,2 Bertrand Czarny,1 Zhi Yang,1 Kingshuk Poddar,2,3 Michal M Dykas,2,3 Abhijeet Patra,2,3 T Venkatesan,2,3 Tomasz Panczyk,4 Chengkuo Lee,5 Giorgia Pastorin1–3 1Department of Pharmacy, National University of Singapore, 2NUS Graduate School for Integrative Sciences and Engineering, Centre for Life Sciences (CeLS, 3NUSNNI-NanoCore, National University of Singapore, Singapore; 4Institute of Catalysis and Surface Chemistry, Polish Academy of Sciences, Cracow, Poland; 5Department of Electrical and Computer Engineering, National University of Singapore, Singapore Abstract: Carbon nanotubes’ (CNTs hollow interior space has been explored for biomedical applications, such as drug repository against undesirable inactivation. To further devise CNTs as smart material for controlled release of cargo molecules, we propose the concept of “gold-carbon nanobottles”. After encapsulating cis-diammineplatinum(II dichloride (cisplatin, CDDP in CNTs, we covalently attached gold nanoparticles (AuNPs at the open-tips of CNTs via different cleavable linkages, namely hydrazine, ester, and disulfide-containing linkages. Compared with our previous study in which more than 80% of CDDP leaked from CNTs in 2 hours, AuNPs were found to significantly decrease such spontaneous release to <40%. In addition, CDDP release from AuNP-capped CNTs via disulfide linkage was selectively enhanced by twofolds in reducing conditions (namely with 1 mM dithiothreitol [DTT], which mimic the intracellular environment. We treated human colon adenocarcinoma cells HCT116 with our CDDP-loaded gold-carbon nanobottles and examined the cell viability using lactate dehydrogenase assay. Interestingly, we found that our nanobottles with cleavable disulfide linkage exerted stronger cytotoxic effect in HCT116 compared with normal human fetal lung fibroblast cells IMR-90. Therefore, we infer that our nanobottles strategy with inbuilt disulfide linkage could

  10. Live-cell imaging of biothiols via thiol/disulfide exchange to trigger the photoinduced electron transfer of gold-nanodot sensor

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Ching-Ping; Wu, Te-Haw; Liu, Chia-Yeh; Lin, Shu-Yi, E-mail: shuyi@nhri.org.tw

    2014-11-07

    Highlights: • The ultrasmall size, PAMAM dendrimer-entrapped Au{sub 8}-clusters were synthesized. • Thiol/disulfide exchange with biothiols to release 2-PyT resulted in quenching. • The sensing platform can detect both low and high molecular weight thiols. • Capable of imaging biothiols including protein thiols in living cells. - Abstract: Biothiols have been reported to involve in intracellular redox-homeostasis against oxidative stress. In this study, a highly selective and sensitive fluorescent probe for sensing biothiols is explored by using an ultrasmall gold nanodot (AuND), the dendrimer-entrapped Au{sub 8}-cluster. This strategy relies upon a thiol/disulfide exchange to trigger the fluorescence change through a photoinduced electron transfer (PET) process between the Au{sub 8}-cluster (as an electron donor) and 2-pyridinethiol (2-PyT) (as an electron acceptor) for sensing biothiols. When 2-PyT is released via the cleavage of disulfide bonds by biothiols, the PET process from the Au{sub 8}-cluster to 2-PyT is initiated, resulting in fluorescence quenching. The fluorescence intensity was found to decrease linearly with glutathione (GSH) concentration (0–1500 μM) at physiological relevant levels and the limit of detection for GSH was 15.4 μM. Compared to most nanoparticle-based fluorescent probes that are limited to detect low molecular weight thiols (LMWTs; i.e., GSH and cysteine), the ultrasmall Au{sub 8}-cluster-based probe exhibited less steric hindrance and can be directly applied in selectively and sensitively detecting both LMWTs and high molecular weight thiols (HMWTs; i.e., protein thiols). Based on such sensing platform, the surface-functionalized Au{sub 8}-cluster has significant promise for use as an efficient nanoprobe for intracellular fluorescence imaging of biothiols including protein thiols in living cells whereas other nanoparticle-based fluorescent probes cannot.

  11. An HLA-B27 Homodimer Specific Antibody Recognizes a Discontinuous Mixed-Disulfide Epitope as Identified by Affinity-Mass Spectrometry

    Science.gov (United States)

    Iuraşcu, Marius-Ionuţ; Marroquin Belaunzanar, Osiris; Cozma, Claudia; Petrausch, Ulf; Renner, Christoph; Przybylski, Michael

    2016-06-01

    HLA-B27 homodimer formation is believed to be a hallmark of HLA-B27 associated spondyloarthritides. Recently, we have generated a homodimer-specific monoclonal antibody (HD6) and have demonstrated that HLA-B27 homodimer complexes are present on monocytes of healthy HLA-B27 gene carriers at low levels, with significantly increased levels at active disease. The capability of the HD6 antibody to discriminate between correctly formed HLA-B27 heterotrimers and pathology-associated homodimers is striking and cannot be explained by the primary structure of HLA-B27. We hypothesized that HD6 accesses a unique epitope and used affinity-mass spectrometry for its identification. The HD6 antibody was immobilized on an activated sepharose affinity column, and HLA-B27 homodimer characterized for affinity. The epitope was identified by proteolytic epitope excision and MALDI mass spectrometry, and shown to comprise a discontinuous Cys-203- 257-Cys mixed-disulfide peptide structure that is not accessible in HLA-B27 heterotrimers due to protection by noncovalently linked β2-microglobulin. The epitope peptides were synthesized by solid phase peptide synthesis, and the two monomeric peptide components, HLA-B27(203-219) and HLA-B27(257-273), as well as the homo- and hetero-dimeric disulfide linked combinations prepared. The affinity binding constants KD towards the antibodies were determined using a surface acoustic wave (SAW) biosensor, and showed the highest affinity with a KD of approximately 40 nM to the HD6 antibody for the (203-219)-SS-(257-273) mixed disulfide epitope.

  12. Cu(II)-disulfide complexes with superoxide dismutase- and catalase-like activities protect mitochondria and whole cells against oxidative stress.

    Science.gov (United States)

    Aliaga, Margarita E; Sandoval-Acuña, Cristián; López-Alarcón, Camilo; Fuentes, Jocelyn; Speisky, Hernan

    2014-10-01

    Mitochondria are a major subcellular site of superoxide (O2(-)) formation. Conditions leading to an uncontrolled production, accumulation and/or conversion of O2(-) into hydrogen peroxide result in an increment in the intramitochondrial oxidative tone which, ultimately leads to the loss of cell viability. Recently, we reported on the ability of a series of Cu(II)-disulfide complexes to act simultaneously as SOD- and catalase-like molecules. In the present study, we addressed the potential of such compounds to protect mitochondria and cells against the oxidative stress and the cytolytic damage induced by diclofenac. Exposure of Caco-2 cells to diclofenac (250µM, 20min) led to a near 80% inhibition of mitochondrial complex I activity and almost doubled the rate of mitochondrial O2(-) production (assessed by Mitosox). A comparable increment was seen in whole cells when the oxidative tone was assessed through the largely hydrogen peroxide-dependent dichlorofluorescein (DCFH) oxidation. The increment in mitochondrial O2(-) production was totally and concentration-dependently prevented by the addition of the complexes formed between Cu(II) and the disulfides of glutathione, homocysteine, or a-dehydro-lipoic acid (20µM each); comparatively, the Cu(II)-cystine complex exerted a weaker protection. A comparable protection pattern was seen at the whole cell level, as these complexes were also effective in preventing the increment in DCFH oxidation. The mitochondrial and whole cell antioxidant protection also translated into a full protection against the cytolytic effects of diclofenac (45min). Results from the present study indicate that the here-tested Cu(II)-disulfides complexes are able to effectively protect cells against the oxidative and the lytic effects of O2(-)-overproducing mitochondria, suggesting a potential for these type of compounds to act as SOD- and catalase-like molecules under oxidative-stress conditions. Supported by FONDECYT #1110018. PMID:26461399

  13. 9.3 kDa Components of the Injected Venom of Conus purpurascens Define a New 5-disulfide Conotoxin Framework

    OpenAIRE

    Möller, Carolina; Marí, Frank

    2011-01-01

    The 83-residue conopeptide (p21a) and its corresponding non-hydroxylated analog were isolated from the injected venom of Conus purpurascens. The complete conopeptide sequences were derived from Edman degradation sequencing of fragments from tryptic, chymotryptic and cyanogen bromide digestions. p21a has a unique, ten-cystine/5-disulfide 7-loop framework with extended 10-residue N-terminus and a 5-residue C-terminus tails, respectively. p21a has a 48% sequence homology with a recently describe...

  14. A supercell, Bloch wave method for calculating low-energy electron reflectivity with applications to free-standing graphene and molybdenum disulfide

    Science.gov (United States)

    McClain, John

    This dissertation reports on a novel theoretical and computational framework for calculating low-energy electron reflectivities from crystalline surfaces and its application to two layered systems of two-dimensional materials, graphene and molybdenum disulfide. The framework provides a simple and efficient approach through the matching of a small set of Fourier components of Bloch wave solutions to the Schrodinger Equation in a slab-in-supercell geometry to incoming and outgoing plane waves on both sides of the supercell. The implementation of this method is described in detail for the calculation of reflectivities in the lowest energy range, for which only specular reflection is allowed. This implementation includes the calculation of reflectivities from beams with normal or off-normal incidence. Two different algorithms are described in the case of off-normal incidence which differ in their dependence on the existence of a symmetry with a mirror plane parallel to the crystal surface. Applications to model potentials in one, two, and three dimensions display consistent results when using different supercell sizes and convergent results with the density of Fourier grids. The design of the Bloch wave matching also allows for the accurate modeling of crystalline slabs through the use of realistic potentials determined via density functional theory. The application of the method to low-energy electron scattering from free-standing systems of a few layers of graphene, including the use of these realistic potentials, demonstrates this ability of the method to accurately model real systems. It reproduces the layer-dependent oscillations found in experimental, normal incidence reflectivity curves for a few layers of graphene grown on silicon carbide. The normal incidence reflectivity curves calculated for slabs consisting of few-layer graphene on 10 layers of nickel show some qualitative agreement with experiment. General incidence reflectivity spectra for free

  15. On the role of protein disulfide isomerase in the retrograde cell transport of secreted phospholipases A2.

    Directory of Open Access Journals (Sweden)

    Jernej Oberčkal

    Full Text Available Following the finding that ammodytoxin (Atx, a neurotoxic secreted phospholipase A2 (sPLA2 in snake venom, binds specifically to protein disulfide isomerase (PDI in vitro we show that these proteins also interact in living rat PC12 cells that are able to internalize this group IIA (GIIA sPLA2. Atx and PDI co-localize in both differentiated and non-differentiated PC12 cells, as shown by fluorescence microscopy. Based on a model of the complex between Atx and yeast PDI (yPDI, a three-dimensional model of the complex between Atx and human PDI (hPDI was constructed. The Atx binding site on hPDI is situated between domains b and b'. Atx interacts hPDI with an extensive area on its interfacial binding surface. The mammalian GIB, GIIA, GV and GX sPLA2s have the same fold as Atx. The first three sPLA2s have been detected intracellularly but not the last one. The models of their complexes with hPDI were constructed by replacement of Atx with the respective mammalian sPLA2 in the Atx-hPDI complex and molecular docking of the structures. According to the generated models, mammalian GIB, GIIA and GV sPLA2s form complexes with hPDI very similar to that with Atx. The contact area between GX sPLA2 and hPDI is however different from that of the other sPLA2s. Heterologous competition of Atx binding to hPDI with GV and GX sPLA2s confirmed the model-based expectation that GV sPLA2 was a more effective inhibitor than GX sPLA2, thus validating our model. The results suggest a role of hPDI in the (pathophysiology of some snake venom and mammalian sPLA2s by assisting the retrograde transport of these molecules from the cell surface. The sPLA2-hPDI model constitutes a valuable tool to facilitate further insights into this process and into the (pathophysiology of sPLA2s in relation to their action intracellularly.

  16. The Garlic Allelochemical Diallyl Disulfide Affects Tomato Root Growth by Influencing Cell Division, Phytohormone Balance and Expansin Gene Expression.

    Science.gov (United States)

    Cheng, Fang; Cheng, Zhihui; Meng, Huanwen; Tang, Xiangwei

    2016-01-01

    Diallyl disulfide (DADS) is a volatile organosulfur compound derived from garlic (Allium sativum L.), and it is known as an allelochemical responsible for the strong allelopathic potential of garlic. The anticancer properties of DADS have been studied in experimental animals and various types of cancer cells, but to date, little is known about its mode of action as an allelochemical at the cytological level. The current research presents further studies on the effects of DADS on tomato (Solanum lycopersicum L.) seed germination, root growth, mitotic index, and cell size in root meristem, as well as the phytohormone levels and expression profile of auxin biosynthesis genes (FZYs), auxin transport genes (SlPINs), and expansin genes (EXPs) in tomato root. The results showed a biphasic, dose-dependent effect on tomato seed germination and root growth under different DADS concentrations. Lower concentrations (0.01-0.62 mM) of DADS significantly promoted root growth, whereas higher levels (6.20-20.67 mM) showed inhibitory effects. Cytological observations showed that the cell length of root meristem was increased and that the mitotic activity of meristematic cells in seedling root tips was enhanced at lower concentrations of DADS. In contrast, DADS at higher concentrations inhibited root growth by affecting both the length and division activity of meristematic cells. However, the cell width of the root meristem was not affected. Additionally, DADS increased the IAA and ZR contents of seedling roots in a dose-dependent manner. The influence on IAA content may be mediated by the up-regulation of FZYs and PINs. Further investigation into the underlying mechanism revealed that the expression levels of tomato EXPs were significantly affected by DADS. The expression levels of EXPB2 and beta-expansin precursor were increased after 3 d, and those of EXP1, EXPB3 and EXLB1 were increased after 5 d of DADS treatment (0.41 mM). This result suggests that tomato root growth may be

  17. The Garlic Allelochemical Diallyl Disulfide Affects Tomato Root Growth by Influencing Cell Division, Phytohormone Balance and Expansin Gene Expression

    Science.gov (United States)

    Cheng, Fang; Cheng, Zhihui; Meng, Huanwen; Tang, Xiangwei

    2016-01-01

    Diallyl disulfide (DADS) is a volatile organosulfur compound derived from garlic (Allium sativum L.), and it is known as an allelochemical responsible for the strong allelopathic potential of garlic. The anticancer properties of DADS have been studied in experimental animals and various types of cancer cells, but to date, little is known about its mode of action as an allelochemical at the cytological level. The current research presents further studies on the effects of DADS on tomato (Solanum lycopersicum L.) seed germination, root growth, mitotic index, and cell size in root meristem, as well as the phytohormone levels and expression profile of auxin biosynthesis genes (FZYs), auxin transport genes (SlPINs), and expansin genes (EXPs) in tomato root. The results showed a biphasic, dose-dependent effect on tomato seed germination and root growth under different DADS concentrations. Lower concentrations (0.01–0.62 mM) of DADS significantly promoted root growth, whereas higher levels (6.20–20.67 mM) showed inhibitory effects. Cytological observations showed that the cell length of root meristem was increased and that the mitotic activity of meristematic cells in seedling root tips was enhanced at lower concentrations of DADS. In contrast, DADS at higher concentrations inhibited root growth by affecting both the length and division activity of meristematic cells. However, the cell width of the root meristem was not affected. Additionally, DADS increased the IAA and ZR contents of seedling roots in a dose-dependent manner. The influence on IAA content may be mediated by the up-regulation of FZYs and PINs. Further investigation into the underlying mechanism revealed that the expression levels of tomato EXPs were significantly affected by DADS. The expression levels of EXPB2 and beta-expansin precursor were increased after 3 d, and those of EXP1, EXPB3 and EXLB1 were increased after 5 d of DADS treatment (0.41 mM). This result suggests that tomato root growth may be

  18. CS2作业工人红细胞膜收缩蛋白初探%Preliminary study on erythrocyte membrane spectrin in workers exposed to carbon disulfide

    Institute of Scientific and Technical Information of China (English)

    简乐; 陈苘

    2000-01-01

    目的 观察作业工人长期接触二硫化碳 (CS2)对其红细胞膜收缩蛋白的影响.方法 气相色谱法测定作业场所CS2浓度;分离16名工人红细胞膜(接触、对照各8人),采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)法分析红细胞膜蛋白谱.结果 车间空气 CS2浓度(TWA)为0~8.98 mg/m3.CS2接触工人红细胞膜收缩蛋白百分含量(34.48%)高于对照工人(26.18%)(t=5.920,P<0.001),CS2浓度(TWA)与收缩蛋白含量呈正相关(r=0.859,R2=0.738,P<0.001),接触指数(EI)与收缩蛋白含量也呈正相关(r=0.713,R2=0.508,P=0.002).其中2名长工龄高浓度CS2接触工人红细胞膜图谱在高相对分子质量区有可疑异常区带形成.结论 长期接触CS2作业工人红细胞膜蛋白电泳图谱出现异常,提示该指标有希望成为CS2接触工人的分子水平接触-效应生物标志物.

  19. Atom economical synthesis of di- and trithiocarbonates by the lithium tert-butoxide catalyzed addition of carbon disulfide to epoxides and thiiranes.

    Science.gov (United States)

    Diebler, J; Spannenberg, A; Werner, T

    2016-08-21

    Alkali metal alkoxides were studied as catalysts for the addition of CS2 to epoxides. A screening of several commercially available alkoxides revealed lithium tert-butoxide as an active and selective catalyst for this reaction. The influence of different reaction parameters as well as the substrate scope under optimized reaction conditions has been studied. Terminal and highly substituted epoxides as well as thiiranes were converted. In total 28 products were prepared and isolated in yields up to 95%. Notably, the reactions were performed under mild conditions without additional solvents. The regio- and stereoselectivity of the reaction has been studied e.g. by converting (R)-styrene and (R)-propylene oxide. Moreover, the test reaction was monitored by (13)C NMR and a plausible mechanism for the conversion of terminal and internal epoxides is given. This proposal is in agreement with the observed regio- and stereoselectivity of the reaction. PMID:27339808

  20. Porous carbons

    Indian Academy of Sciences (India)

    Satish M Manocha

    2003-02-01

    Carbon in dense as well as porous solid form is used in a variety of applications. Activated porous carbons are made through pyrolysis and activation of carbonaceous natural as well as synthetic precursors. Pyrolysed woods replicate the structure of original wood but as such possess very low surface areas and poor adsorption capacities. On activation, these exhibit increased adsorption volumes of 0.5–0.8 cm3 /gm and surface areas of 700–1800 m2 /gm depending on activation conditions, whether physical or chemical. Former carbons possess mixed pore size distribution while chemically activated carbons predominantly possess micropores. Thus, these carbons can be used for adsorption of wide distributions of molecules from gas to liquid. The molecular adsorption within the pores is due to single layer or multilayer molecule deposition at the pore walls and hence results in different types of adsorption isotherm. On the other hand, activated carbon fibres with controlled microporous structure and surface area in the range of 2500 m2 /gm can be developed by controlled pyrolysis and physical activation of amorphous carbon fibres. Active carbon fibres with unmatchable pore structure and surface characteristics are present and futuristic porous materials for a number of applications from pollution control to energy storage.

  1. A thiol-disulfide oxidoreductase of the Gram-positive pathogen Corynebacterium diphtheriae is essential for viability, pilus assembly, toxin production and virulence

    Science.gov (United States)

    Reardon-Robinson, Melissa E.; Osipiuk, Jerzy; Jooya, Neda; Chang, Chungyu; Joachimiak, Andrzej; Das, Asis; Ton-That, Hung

    2016-01-01

    Summary The Gram-positive pathogen Corynebacterium diphtheriae exports through the Sec apparatus many extracellular proteins that include the key virulence factors diphtheria toxin and the adhesive pili. How these proteins attain their native conformations after translocation as unfolded precursors remains elusive. The fact that the majority of these exported proteins contain multiple cysteine residues and that several membrane-bound oxidoreductases are encoded in the corynebacterial genome suggests the existence of an oxidative protein-folding pathway in this organism. Here we show that the shaft pilin SpaA harbors a disulfide bond in vivo and alanine substitution of these cysteines abrogates SpaA polymerization and leads to the secretion of degraded SpaA peptides. We then identified a thiol-disulfide oxidoreductase (MdbA), whose structure exhibits a conserved thioredoxin-like domain with a CPHC active site. Remarkably, deletion of mdbA results in a severe temperature-sensitive cell division phenotype. This mutant also fails to assemble pilus structures and is greatly defective in toxin production. Consistent with these defects, the ΔmdbA mutant is attenuated in a guinea pig model of diphtheritic toxemia. Given its diverse cellular functions in cell division, pilus assembly and toxin production, we propose that MdbA is a component of the general oxidative folding machine in C. diphtheriae. PMID:26294390

  2. Direct, simple derivatization of disulfide bonds in proteins with organic mercury in alkaline medium without any chemical pre-reducing agents

    Energy Technology Data Exchange (ETDEWEB)

    Campanella, Beatrice; Onor, Massimo [National Research Council of Italy, C.N.R., Istituto di Chimica dei Composti Organo Metallici-ICCOM- UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa (Italy); Ferrari, Carlo [National Research Council of Italy, C.N.R., Istituto Nazionale di Ottica, INO-UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa (Italy); D’Ulivo, Alessandro [National Research Council of Italy, C.N.R., Istituto di Chimica dei Composti Organo Metallici-ICCOM- UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa (Italy); Bramanti, Emilia, E-mail: bramanti@pi.iccom.cnr.it [National Research Council of Italy, C.N.R., Istituto di Chimica dei Composti Organo Metallici-ICCOM- UOS Pisa, Area di Ricerca, Via G. Moruzzi 1, 56124 Pisa (Italy)

    2014-09-16

    Highlights: • A simple procedure for the derivatization of proteins disulfide bonds. • Cysteine groups in several proteins derivatised with pHMB in alkaline media. • 75–100% labelling of cysteines in proteins with pHMB. - Abstract: In this work we have studied the derivatization of protein disulfide bonds with p-Hydroxymercurybenzoate (pHMB) in strong alkaline medium without any preliminary reduction. The reaction has been followed by the determination of the protein–pHMB complex using size exclusion chromatography coupled to a microwave/UV mercury oxidation system for the on-line oxidation of free and protein-complexed pHMB and atomic fluorescence spectrometry (SEC–CVG–AFS) detection. The reaction has been optimized by an experimental design using lysozyme as a model protein and applied to several thiolic proteins. The proposed method reports, for the first time, that it is possible to label 75–100% cysteines of proteins and, thus, to determine thiolic proteins without the need of any reducing step to obtain reduced -SH groups before mercury labelling. We obtained a detection limit of 100 nmol L{sup −1} based on a signal-to-noise ratio of 3 for unbound and complexed pHMB, corresponding to a detection limit of proteins ranged between 3 and 360 nmol L{sup −1}, depending on the number of cysteines in the protein sequence.

  3. Effects of garlic oil and two of its major organosulfur compounds, diallyl disulfide and diallyl trisulfide, on intestinal damage in rats injected with endotoxin

    International Nuclear Information System (INIS)

    Garlic and its active components are known to possess antioxidant and antiinflammatory effects. The present study investigated the effects of garlic oil and its organosulfur compounds on endotoxin-induced intestinal mucosal damage. Wistar rats received by gavage 50 or 200 mg/kg body weight garlic oil (GO), 0.5 mmol/kg body weight diallyl disulfide or diallyl trisulfide, or the vehicle (corn oil; 2 ml/kg body weight) every other day for 2 weeks before being injected with endotoxin (i.p., 5 mg/kg body weight). Control rats were administered with corn oil and were injected with sterile saline. Samples for the measurement of proinflammatory cytokines were collected 3 h after injection, and all other samples were collected 18 h after injection. The low dose of GO suppressed endotoxin-induced inducible nitric oxide synthase (iNOS) activity, ulceration, and apoptosis in the intestinal mucosa (P < 0.05). The high dose of GO significantly lowered the peripheral level of nitrate/nitrite and endotoxin-induced iNOS activity in the intestinal mucosa (P < 0.05) but worsened intestinal mucosal damage accompanied by elevated peripheral proinflammatory cytokines. Diallyl trisulfide but not diallyl disulfide showed similar toxic effect as that of high-dose GO. These results suggest the preventive effect and possible toxicity of garlic oil and its organosulfur compounds in endotoxin-induced systemic inflammation and intestinal damage

  4. Crystal structures of mono- and bi-specific diabodies and reduction of their structural flexibility by introduction of disulfide bridges at the Fv interface

    Science.gov (United States)

    Kim, Jin Hong; Song, Dong Hyun; Youn, Suk-Jun; Kim, Ji Won; Cho, Geunyoung; Kim, Sun Chang; Lee, Hayyoung; Jin, Mi Sun; Lee, Jie-Oh

    2016-01-01

    Building a sophisticated protein nano-assembly requires a method for linking protein components in a predictable and stable structure. Diabodies are engineered antibody fragments that are composed of two Fv domains connected by short peptide linkers. They are attractive candidates for mediators in assembling protein nano-structures because they can simultaneously bind to two different proteins and are rigid enough to be crystallized. However, comparison of previous crystal structures demonstrates that there is substantial structural diversity in the Fv interface region of diabodies and, therefore, reliable prediction of its structure is not trivial. Here, we present the crystal structures of ten mono- and bi-specific diabodies. We found that changing an arginine residue in the Fv interface to threonine greatly reduced the structural diversity of diabodies. We also found that one of the bispecific diabodies underwent an unexpected process of chain swapping yielding a non-functional monospecific diabody. In order to further reduce structural flexibility and prevent chain shuffling, we introduced disulfide bridges in the Fv interface regions. The disulfide-bridged diabodies have rigid and predictable structures and may have applications in crystallizing proteins, analyzing cryo-electron microscopic images and building protein nano-assemblies. PMID:27682821

  5. A sensitive electrochemical aptasensor based on palladium nanoparticles decorated graphene-molybdenum disulfide flower-like nanocomposites and enzymatic signal amplification.

    Science.gov (United States)

    Jing, Pei; Yi, Huayu; Xue, Shuyan; Chai, Yaqin; Yuan, Ruo; Xu, Wenju

    2015-01-01

    In the present study, with the aggregated advantages of graphene and molybdenum disulfide (MoS2), we prepared poly(diallyldimethylammonium chloride)-graphene/molybdenum disulfide (PDDA-G-MoS2) nanocomposites with flower-like structure, large surface area and excellent conductivity. Furthermore, an advanced sandwich-type electrochemical assay for sensitive detection of thrombin (TB) was fabricated using palladium nanoparticles decorated PDDA-G-MoS2 (PdNPs/PDDA-G-MoS2) as nanocarriers, which were functionalized by hemin/G-quadruplex, glucose oxidase (GOD), and toluidine blue (Tb) as redox probes. The signal amplification strategy was achieved as follows: Firstly, the immobilized GOD could effectively catalyze the oxidation of glucose to gluconolactone, coupling with the reduction of the dissolved oxygen to H2O2. Then, both PdNPs and hemin/G-quadruplex acting as hydrogen peroxide (HRP)-mimicking enzyme could further catalyze the reduction of H2O2, resulting in significant electrochemical signal amplification. So the proposed aptasensor showed high sensitivity with a wide dynamic linear range of 0.0001 to 40 nM and a relatively low detection limit of 0.062 pM for TB determination. The strategy showed huge potential of application in protein detection and disease diagnosis.

  6. The coupled effects of environmental composition, temperature and contact size-scale on the tribology of molybdenum disulfide

    Science.gov (United States)

    Khare, Harmandeep S.

    Liquid lubricants are precluded in an exceedingly large number of consumer as well as extreme applications as a means to reduce friction and wear at the sliding interface of two bodies. The extraterrestrial environment is one such example of an extreme environment which has motivated the development of advanced solid lubricant materials. Mechanical systems for space require fabrication, assembly, transportation and testing on earth before launch and deployment. Solid lubricants for space are expected to not only operate efficiently in the hard vacuum of space but also withstand interactions with moisture or oxygen during the terrestrial storage, transportation and assembly prior to deployment and launch. Molybdenum disulfide (MoS2) is considered the gold standard in solid lubricants for space due to its excellent tribological properties in ultra-high vacuum. However in the presence of environmental species such as water and oxygen or at elevated temperatures, the lubricity and endurance of MoS2 is severely limited. Past studies have offered several hypotheses for the breakdown of lubrication of MoS2 under the influence of water and oxygen, although exact mechanisms remain unknown. Furthermore, it is unclear if temperature acts as a driver solely for oxidation or for thermally activated slip and thermally activated desorption as well. The answers to these questions are of fundamental importance to improving the reliability of existing MoS2-based solid lubricants for space, as well as for guiding the design of advanced lamellar solid lubricant coatings. This dissertation aims to elucidate: (1) the role of water on MoS2 oxidation, (2) the role of water on MoS2 friction, (3) the role of oxygen on MoS2 friction, (4) the contribution of thermal activation to ambient-temperature friction, and (5) effects of length-scale. The results of this study showed that water does not cause oxidation of MoS2. Water increases ambient-temperature friction of MoS2 directly through a

  7. Calcium Carbonate

    Science.gov (United States)

    ... before being swallowed; do not swallow them whole. Drink a full glass of water after taking either the regular or chewable tablets or capsules. Some liquid forms of calcium carbonate must be shaken well before use.Do not ...

  8. Carbon Stars

    Indian Academy of Sciences (India)

    T. Lloyd Evans

    2010-12-01

    In this paper, the present state of knowledge of the carbon stars is discussed. Particular attention is given to issues of classification, evolution, variability, populations in our own and other galaxies, and circumstellar material.

  9. Rv2969c, essential for optimal growth in Mycobacterium tuberculosis, is a DsbA-like enzyme that interacts with VKOR-derived peptides and has atypical features of DsbA-like disulfide oxidases

    Energy Technology Data Exchange (ETDEWEB)

    Premkumar, Lakshmanane, E-mail: p.lakshmanane@imb.uq.edu.au; Heras, Begoña; Duprez, Wilko; Walden, Patricia; Halili, Maria; Kurth, Fabian; Fairlie, David P.; Martin, Jennifer L., E-mail: p.lakshmanane@imb.uq.edu.au [University of Queensland, St Lucia, QLD 4067 (Australia)

    2013-10-01

    The gene product of M. tuberculosis Rv2969c is shown to be a disulfide oxidase enzyme that has a canonical DsbA-like fold with novel structural and functional characteristics. The bacterial disulfide machinery is an attractive molecular target for developing new antibacterials because it is required for the production of multiple virulence factors. The archetypal disulfide oxidase proteins in Escherichia coli (Ec) are DsbA and DsbB, which together form a functional unit: DsbA introduces disulfides into folding proteins and DsbB reoxidizes DsbA to maintain it in the active form. In Mycobacterium tuberculosis (Mtb), no DsbB homologue is encoded but a functionally similar but structurally divergent protein, MtbVKOR, has been identified. Here, the Mtb protein Rv2969c is investigated and it is shown that it is the DsbA-like partner protein of MtbVKOR. It is found that it has the characteristic redox features of a DsbA-like protein: a highly acidic catalytic cysteine, a highly oxidizing potential and a destabilizing active-site disulfide bond. Rv2969c also has peptide-oxidizing activity and recognizes peptide segments derived from the periplasmic loops of MtbVKOR. Unlike the archetypal EcDsbA enzyme, Rv2969c has little or no activity in disulfide-reducing and disulfide-isomerase assays. The crystal structure of Rv2969c reveals a canonical DsbA fold comprising a thioredoxin domain with an embedded helical domain. However, Rv2969c diverges considerably from other DsbAs, including having an additional C-terminal helix (H8) that may restrain the mobility of the catalytic helix H1. The enzyme is also characterized by a very shallow hydrophobic binding surface and a negative electrostatic surface potential surrounding the catalytic cysteine. The structure of Rv2969c was also used to model the structure of a paralogous DsbA-like domain of the Ser/Thr protein kinase PknE. Together, these results show that Rv2969c is a DsbA-like protein with unique properties and a limited

  10. Conformational analysis of large and highly disulfide-stabilized proteins by integrating online electrochemical reduction into an optimized H/D exchange mass spectrometry workflow

    DEFF Research Database (Denmark)

    Trabjerg, Esben; Jakobsen, Rasmus Uffe; Mysling, Simon;

    2015-01-01

    solution (e.g. TCEP). The chemical reduction, however, is severely limited under quenched conditions due to a narrow time window as well as low pH and temperature. Here, we demonstrate the real-world applicability of integrating electrochemical reduction into an online HDX-MS workflow. We have optimized...... the electrochemical reduction efficiency during HDX-MS analysis of two particularly challenging disulfide stabilized proteins: a therapeutic IgG1-antibody and Nerve Growth Factor-β (NGF). Several different parameters (flow rate, applied square wave potential as well as the type of labeling- and quench buffer) were...... investigated, and the optimized workflow increased the sequence coverage of NGF from 46% with chemical reduction to 99%, when electrochemical reduction was applied. Additionally, the optimized workflow also enabled a similar high sequence coverage of 96% and 87% for the heavy and light chain of the IgG1...

  11. Molecular Design of Crown Ethers. 191. Synthesis of Novel Disulfide- and Diselenide-Bridged Bis (benzo-12-crown-4)s and their Ag+ Selective Electrode Properties

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Two novel heteroatom-bridged bis (benzo-12-crown-4 ether)s, i.e. bis [2-nitro-4,5-(1,4,7,10-tetraoxadecamethylene)phenyl] disulfide 1 and diselenide 2, have been synthesized. X-ray crystallographic structure was obtained for 1. Ion selective electrodes (ISE) for Ag+, containing 1 and 2 in PVC membrane as neutral carriers, were prepared, and their selectivity coefficients for Ag+ (KpolAg,M) were determined against other heavy metal ions, alkali and alkaline-earth metal ions, and ammonium ion. These ISEs showed excellent Ag+ selectivities, log (KpolAg,M) -3.8, against most of the interfering cations examined, except for Hg+.

  12. Cell-Penetrating Poly(disulfide) Assisted Intracellular Delivery of Mesoporous Silica Nanoparticles for Inhibition of miR-21 Function and Detection of Subsequent Therapeutic Effects.

    Science.gov (United States)

    Yu, Changmin; Qian, Linghui; Ge, Jingyan; Fu, Jiaqi; Yuan, Peiyan; Yao, Samantha C L; Yao, Shao Q

    2016-08-01

    The design of drug delivery systems capable of minimal endolysosomal trapping, controlled drug release, and real-time monitoring of drug effect is highly desirable for personalized medicine. Herein, by using mesoporous silica nanoparticles (MSNs) coated with cell-penetrating poly(disulfide)s and a fluorogenic apoptosis-detecting peptide (DEVD-AAN), we have developed a platform that could be uptaken rapidly by mammalian cells via endocytosis-independent pathways. Subsequent loading of these MSNs with small molecule inhibitors and antisense oligonucleotides resulted in intracellular release of these drugs, leading to combination inhibition of endogenous miR-21 activities which was immediately detectable by the MSN surface-coated peptide using two-photon fluorescence microscopy. PMID:27325284

  13. Influence of Disulfide-Stabilized Structure on the Specificity of Helper T-Cell and Antibody Responses to HIV Envelope Glycoprotein gp120▿ †

    Science.gov (United States)

    Mirano-Bascos, Denise; Steede, N. Kalaya; Robinson, James E.; Landry, Samuel J.

    2010-01-01

    CD4+ helper T cells specific for human immunodeficiency virus type 1 (HIV-1) are associated with control of viremia. Nevertheless, vaccines have had limited effectiveness thus far, in part because sequence variability and other structural features of the HIV envelope glycoprotein deflect the immune response. Previous studies indicated that CD4+ T-cell epitope dominance is controlled by antigen three-dimensional structure through its influence on antigen processing and presentation. In this work, three disulfide bonds in the outer domain of gp120 were individually deleted in order to destabilize the local three-dimensional structure and enhance the presentation of nearby weakly immunogenic epitopes. However, upon immunization of groups of BALB/c mice, the CD4+ T-cell response was broadly reduced for all three variants, and distinct epitope profiles emerged. For one variant, antibody titers were sharply increased, and the antibody exhibited significant CD4-blocking activity. PMID:20089653

  14. Prokaryotic soluble overexpression and purification of bioactive human growth hormone by fusion to thioredoxin, maltose binding protein, and protein disulfide isomerase.

    Directory of Open Access Journals (Sweden)

    Minh Tan Nguyen

    Full Text Available Human growth hormone (hGH is synthesized by somatotroph cells of the anterior pituitary gland and induces cell proliferation and growth. This protein has been approved for the treatment of various conditions, including hGH deficiency, chronic renal failure, and Turner syndrome. Efficient production of hGH in Escherichia coli (E. coli has proven difficult because the E. coli-expressed hormone tends to aggregate and form inclusion bodies, resulting in poor solubility. In this study, seven N-terminal fusion partners, hexahistidine (His6, thioredoxin (Trx, glutathione S-transferase (GST, maltose-binding protein (MBP, N-utilization substance protein A (NusA, protein disulfide bond isomerase (PDI, and the b'a' domain of PDI (PDIb'a', were tested for soluble overexpression of codon-optimized hGH in E. coli. We found that MBP and hPDI tags significantly increased the solubility of the hormone. In addition, lowering the expression temperature to 18°C also dramatically increased the solubility of all the fusion proteins. We purified hGH from MBP-, PDIb'a'-, or Trx-tagged hGH expressed at 18°C in E. coli using simple chromatographic techniques and compared the final purity, yield, and activity of hGH to assess the impact of each partner protein. Purified hGH was highly pure on silver-stained gel and contained very low levels of endotoxin. On average, ∼37 mg, ∼12 mg, and ∼7 mg of hGH were obtained from 500 mL-cell cultures of Trx-hGH, MBP-hGH, and PDIb'a'-hGH, respectively. Subsequently, hGH was analyzed using mass spectroscopy to confirm the presence of two intra-molecular disulfide bonds. The bioactivity of purified hGHs was demonstrated using Nb2-11 cell.

  15. In Silico Identification of Protein Disulfide Isomerase Gene Families in the De Novo Assembled Transcriptomes of Four Different Species of the Genus Conus.

    Science.gov (United States)

    Figueroa-Montiel, Andrea; Ramos, Marco A; Mares, Rosa E; Dueñas, Salvador; Pimienta, Genaro; Ortiz, Ernesto; Possani, Lourival D; Licea-Navarro, Alexei F

    2016-01-01

    Small peptides isolated from the venom of the marine snails belonging to the genus Conus have been largely studied because of their therapeutic value. These peptides can be classified in two groups. The largest one is composed by peptides rich in disulfide bonds, and referred to as conotoxins. Despite the importance of conotoxins given their pharmacology value, little is known about the protein disulfide isomerase (PDI) enzymes that are required to catalyze their correct folding. To discover the PDIs that may participate in the folding and structural maturation of conotoxins, the transcriptomes of the venom duct of four different species of Conus from the peninsula of Baja California (Mexico) were assembled. Complementary DNA (cDNA) libraries were constructed for each species and sequenced using a Genome Analyzer Illumina platform. The raw RNA-seq data was converted into transcript sequences using Trinity, a de novo assembler that allows the grouping of reads into contigs without a reference genome. An N50 value of 605 was established as a reference for future assemblies of Conus transcriptomes using this software. Transdecoder was used to extract likely coding sequences from Trinity transcripts, and PDI-specific sequence motif "APWCGHCK" was used to capture potential PDIs. An in silico analysis was performed to characterize the group of PDI protein sequences encoded by the duct-transcriptome of each species. The computational approach entailed a structural homology characterization, based on the presence of functional Thioredoxin-like domains. Four different PDI families were characterized, which are constituted by a total of 41 different gene sequences. The sequences had an average of 65% identity with other PDIs. Using MODELLER 9.14, the homology-based three-dimensional structure prediction of a subset of the sequences reported, showed the expected thioredoxin fold which was confirmed by a "simulated annealing" method.

  16. Vibrational analysis of amino acids and short peptides in aqueous media. V. The effect of the disulfide bridge on the structural features of the peptide hormone somatostatin-14.

    Science.gov (United States)

    Hernández, Belén; Carelli, Claude; Coïc, Yves-Marie; De Coninck, Joël; Ghomi, Mahmoud

    2009-09-24

    To emphasize the role played by the S-S bridge in the structural features of somatostatin-14 (SST-14), newly recorded CD and Raman spectra of this cyclic peptide and its open analogue obtained by Cys-->Ser substitution are presented. CD spectra of both peptides recorded in aqueous solutions in the 100-500 microM concentration range are strikingly similar. They reveal principally that random conformers constitute the major population in both peptides. Consequently, the S-S bridge has no structuring effect at submillimolar concentrations. In methanol, the CD spectrum of somatostatin-14 keeps globally the same spectral shape as that observed in water, whereas its open analogue presents a major population of helical conformers. Raman spectra recorded as a function of peptide concentration (5-20 mM) and also in the presence of 150 mM NaCl provide valuable conformational information. All Raman spectra present a mixture of random and beta-hairpin structures for both cyclic and open peptides. More importantly, the presence or the absence of the disulfide bridge does not seem to influence considerably different populations of secondary structures within this range of concentrations. CD and Raman data obtained in the submillimolar and millimolar ranges of concentrations, respectively, lead us to accept the idea that SST-14 monomers aggregate upon increasing concentration, thus stabilizing beta-hairpin conformations in solution. However, even at high concentrations, random conformers do not disappear. Raman spectra of SST-14 also reveal a concentration effect on the flexibility of the S-S linkage and consequently on that of its cyclic part. In conclusion, although the disulfide linkage does not seem to markedly influence the SST-14 conformational features in aqueous solutions, its presence seems to be necessary to ensure the flexibility of the cyclic part of this peptide and to maintain its closed structure in lower dielectric constant environments.

  17. Development of Molybdenum Disulfide Intercalation Compounds%二硫化钼夹层化合物的研究进展

    Institute of Scientific and Technical Information of China (English)

    邵鑫; 田军; 邢玉梅; 薛群基; 马春林

    2001-01-01

      Molybdenum disulfide intercalation compounds (MoS2-IC), which attract much attention for its outstanding physical properties, are a kind of new functional nano-materials with wide prospects for application. With the advancement of modern analytical methods, the basic study and applied study closely related to MoS2-IC are also developed. In order to promote the research and development of MoS2-IC, the preparations, structures, properties and applications of MoS2-IC are reviewed in this article. In addition, the principle and the research direction of the development of molybdenum disulfide intercalation compounds were presented.%  二硫化钼夹层化合物是一类极具诱人前景的新功能材料,其以优异的物理性能引起人们极大的兴趣。随着现代分析测试手段的不断进步,有关二硫化钼夹层化合物的基础研究和应用研究也得到了发展。该文对二硫化钼夹层化合物的制备、结构、性能及应用前景等方面的研究状况作了综合介绍与评述,并指出了二硫化钼夹层化合物的研究发展方向。

  18. Study of the thiol/disulfide redox systems of the anaerobe Desulfovibrio vulgaris points out pyruvate:ferredoxin oxidoreductase as a new target for thioredoxin 1.

    Science.gov (United States)

    Pieulle, Laetitia; Stocker, Pierre; Vinay, Manon; Nouailler, Matthieu; Vita, Nicolas; Brasseur, Gaël; Garcin, Edwige; Sebban-Kreuzer, Corinne; Dolla, Alain

    2011-03-11

    Sulfate reducers have developed a multifaceted adaptative strategy to survive against oxidative stresses. Along with this oxidative stress response, we recently characterized an elegant reversible disulfide bond-dependent protective mechanism in the pyruvate:ferredoxin oxidoreductase (PFOR) of various Desulfovibrio species. Here, we searched for thiol redox systems involved in this mechanism. Using thiol fluorescent labeling, we show that glutathione is not the major thiol/disulfide balance-controlling compound in four different Desulfovibrio species and that no other plentiful low molecular weight thiol can be detected. Enzymatic analyses of two thioredoxins (Trxs) and three thioredoxin reductases allow us to propose the existence of two independent Trx systems in Desulfovibrio vulgaris Hildenborough (DvH). The TR1/Trx1 system corresponds to the typical bacterial Trx system. We measured a TR1 apparent K(m) value for Trx1 of 8.9 μM. Moreover, our results showed that activity of TR1 was NADPH-dependent. The second system named TR3/Trx3 corresponds to an unconventional Trx system as TR3 used preferentially NADH (K(m) for NADPH, 743 μM; K(m) for NADH, 5.6 μM), and Trx3 was unable to reduce insulin. The K(m) value of TR3 for Trx3 was 1.12 μM. In vitro experiments demonstrated that the TR1/Trx1 system was the only one able to reactivate the oxygen-protected form of Desulfovibrio africanus PFOR. Moreover, ex vivo pulldown assays using the mutant Trx1(C33S) as bait allowed us to capture PFOR from the DvH extract. Altogether, these data demonstrate that PFOR is a new target for Trx1, which is probably involved in the protective switch mechanism of the enzyme. PMID:21199874

  19. A sensitive electrochemical aptasensor based on palladium nanoparticles decorated graphene–molybdenum disulfide flower-like nanocomposites and enzymatic signal amplification

    Energy Technology Data Exchange (ETDEWEB)

    Jing, Pei; Yi, Huayu; Xue, Shuyan; Chai, Yaqin; Yuan, Ruo; Xu, Wenju, E-mail: xwju@swu.edu.cn

    2015-01-01

    Highlights: • PDDA–G–MoS{sub 2} nanoflowers were firstly used for the fabrication of thrombin aptasensor. • MoS{sub 2} was adopted to enhance the surface area of graphene and accelerate the electron transfer. • GOD, PdNPs and hemin/G-quadruplex could amplify the electrochemical signal through synergetic catalysis. • The proposed aptasensor displayed an improved sensitivity. - Abstract: In the present study, with the aggregated advantages of graphene and molybdenum disulfide (MoS{sub 2}), we prepared poly(diallyldimethylammonium chloride)–graphene/molybdenum disulfide (PDDA–G–MoS{sub 2}) nanocomposites with flower-like structure, large surface area and excellent conductivity. Furthermore, an advanced sandwich-type electrochemical assay for sensitive detection of thrombin (TB) was fabricated using palladium nanoparticles decorated PDDA–G–MoS{sub 2} (PdNPs/PDDA–G–MoS{sub 2}) as nanocarriers, which were functionalized by hemin/G-quadruplex, glucose oxidase (GOD), and toluidine blue (Tb) as redox probes. The signal amplification strategy was achieved as follows: Firstly, the immobilized GOD could effectively catalyze the oxidation of glucose to gluconolactone, coupling with the reduction of the dissolved oxygen to H{sub 2}O{sub 2}. Then, both PdNPs and hemin/G-quadruplex acting as hydrogen peroxide (HRP)-mimicking enzyme could further catalyze the reduction of H{sub 2}O{sub 2}, resulting in significant electrochemical signal amplification. So the proposed aptasensor showed high sensitivity with a wide dynamic linear range of 0.0001 to 40 nM and a relatively low detection limit of 0.062 pM for TB determination. The strategy showed huge potential of application in protein detection and disease diagnosis.

  20. Xanthates and trithiocarbonates strongly inhibit carbonic anhydrases and show antiglaucoma effects in vivo.

    Science.gov (United States)

    Carta, Fabrizio; Akdemir, Atilla; Scozzafava, Andrea; Masini, Emanuela; Supuran, Claudiu T

    2013-06-13

    Dithiocarbamates (DTCs) were recently discovered as carbonic anhydrase (CA, EC 4.2.1.1) inhibitors. A series of xanthates and a trithiocarbonate, structurally related to the DTCs, were prepared by reaction of alcohols/thiols with carbon disulfide in the presence of bases. These compounds were tested for the inhibition of four human (h) isoforms, hCA I, II, IX, and XII, involved in pathologies such as glaucoma (CA II and XII) or cancer (CA IX). Several low nanomolar xanthate/trithiocarbonate inhibitors targeting these CAs were detected. A docking study of some xanthates within the CA II active site showed that these compounds bind in a similar manner with the dithiocarbamates, coordinating monodentately to the Zn(II) ion from the enzyme active site. Several xanthates showed potent intraocular pressure lowering activity in two animal models of glaucoma via the topical administration. Xanthates and thioxanthates represent two novel, promising classes of CA inhibitors. PMID:23647428

  1. Effects of Divalent Cations and Disulfide Bond Reducing Agents on Specific Binding of Growth Hormone to Liver Membrane Receptors from Snakehead Fish (Ophiocephalus argus, Cantor).

    Science.gov (United States)

    Sun, Xun; Zhang, Xin-Na; Zhu, Shang-Quan; Zheng, Han-Qi

    2000-01-01

    Divalent cations, Ca(2 ), Mg(2 ) and Mn(2 ) enhance the binding of bream growth hormone (brGH) to snakehead fish liver membrane, and their optimum concentration was found to be 8 12 mmol/L, at which Ca(2 ), Mg(2 ) and Mn(2 ) could increase, respectively, the specific binding to 230%, 180%, and 200%, compared with the binding in the absence of ions. The Eadie-Scatchard plot was used for the dynamic analysis of the Ca(2 ) binding site. A low affinity Ca(2 ) binding site was found in the GH-receptor complex with K(m)=0.384 mmol/L, and the affinity constant (K(a)) was increased from 1.045x10(9) L.mol(-1) to 1.295x10(9) L.mol(-1) by the addition of 10 mmol/L CaCl(2). The effects of disulfide bond reducing agents, DTT and ME, on (125)I-brGH binding to growth hormone receptor (GHR) on snakehead fish liver memebrane were also analyzed. The addition of 0.1 20 mmol/L DTT or 0.01% 1% ME to the radioreceptor assay system caused a significant dose dependent increase in the specific binding for (125)I-brGH. In the presence of 0.8 mmol/L DTT or 0.08% ME, the specific binding of (125)I-brGH was increased from 10.2% to 15.5% and 13.2% respectively, and the affinity constant was also increased from 1.265x10(9) L.mol(-1) to 2.185x10(9) L.mol(-1) and 1.625x10(9) L.mol(-1), respectively but no changes in the binding capacity were observed. Further studies showed that the effects of reductants on the specific binding of brGH were due in part to the ligand itself and in part to GHR. In addition, it was observed that one of the three disulfide bonds of brGH could be reduced by 0.8 mmol/L DTT.

  2. The Human Sodium-Glucose Cotransporter (hSGLT1) Is a Disulfide-Bridged Homodimer with a Re-Entrant C-Terminal Loop

    Science.gov (United States)

    Sasseville, Louis J.; Morin, Michael; Coady, Michael J.; Blunck, Rikard; Lapointe, Jean-Yves

    2016-01-01

    Na-coupled cotransporters are proteins that use the trans-membrane electrochemical gradient of Na to activate the transport of a second solute. The sodium-glucose cotransporter 1 (SGLT1) constitutes a well-studied prototype of this transport mechanism but essential molecular characteristics, namely its quaternary structure and the exact arrangement of the C-terminal transmembrane segments, are still debated. After expression in Xenopus oocytes, human SGLT1 molecules (hSGLT1) were labelled on an externally accessible cysteine residue with a thiol-reactive fluorophore (tetramethylrhodamine-C5-maleimide, TMR). Addition of dipicrylamine (DPA, a negatively-charged amphiphatic fluorescence “quencher”) to the fluorescently-labelled oocytes is used to quench the fluorescence originating from hSGLT1 in a voltage-dependent manner. Using this arrangement with a cysteine residue introduced at position 624 in the loop between transmembrane segments 12 and 13, the voltage-dependent fluorescence signal clearly indicated that this portion of the 12–13 loop is located on the external side of the membrane. As the 12–13 loop begins on the intracellular side of the membrane, this suggests that the 12–13 loop is re-entrant. Using fluorescence resonance energy transfer (FRET), we observed that different hSGLT1 molecules are within molecular distances from each other suggesting a multimeric complex arrangement. In agreement with this conclusion, a western blot analysis showed that hSGLT1 migrates as either a monomer or a dimer in reducing and non-reducing conditions, respectively. A systematic mutational study of endogenous cysteine residues in hSGLT1 showed that a disulfide bridge is formed between the C355 residues of two neighbouring hSGLT1 molecules. It is concluded that, 1) hSGLT1 is expressed as a disulfide bridged homodimer via C355 and that 2) a portion of the intracellular 12–13 loop is re-entrant and readily accessible from the extracellular milieu. PMID:27137918

  3. The Human Sodium-Glucose Cotransporter (hSGLT1 Is a Disulfide-Bridged Homodimer with a Re-Entrant C-Terminal Loop.

    Directory of Open Access Journals (Sweden)

    Louis J Sasseville

    Full Text Available Na-coupled cotransporters are proteins that use the trans-membrane electrochemical gradient of Na to activate the transport of a second solute. The sodium-glucose cotransporter 1 (SGLT1 constitutes a well-studied prototype of this transport mechanism but essential molecular characteristics, namely its quaternary structure and the exact arrangement of the C-terminal transmembrane segments, are still debated. After expression in Xenopus oocytes, human SGLT1 molecules (hSGLT1 were labelled on an externally accessible cysteine residue with a thiol-reactive fluorophore (tetramethylrhodamine-C5-maleimide, TMR. Addition of dipicrylamine (DPA, a negatively-charged amphiphatic fluorescence "quencher" to the fluorescently-labelled oocytes is used to quench the fluorescence originating from hSGLT1 in a voltage-dependent manner. Using this arrangement with a cysteine residue introduced at position 624 in the loop between transmembrane segments 12 and 13, the voltage-dependent fluorescence signal clearly indicated that this portion of the 12-13 loop is located on the external side of the membrane. As the 12-13 loop begins on the intracellular side of the membrane, this suggests that the 12-13 loop is re-entrant. Using fluorescence resonance energy transfer (FRET, we observed that different hSGLT1 molecules are within molecular distances from each other suggesting a multimeric complex arrangement. In agreement with this conclusion, a western blot analysis showed that hSGLT1 migrates as either a monomer or a dimer in reducing and non-reducing conditions, respectively. A systematic mutational study of endogenous cysteine residues in hSGLT1 showed that a disulfide bridge is formed between the C355 residues of two neighbouring hSGLT1 molecules. It is concluded that, 1 hSGLT1 is expressed as a disulfide bridged homodimer via C355 and that 2 a portion of the intracellular 12-13 loop is re-entrant and readily accessible from the extracellular milieu.

  4. The Human Sodium-Glucose Cotransporter (hSGLT1) Is a Disulfide-Bridged Homodimer with a Re-Entrant C-Terminal Loop.

    Science.gov (United States)

    Sasseville, Louis J; Morin, Michael; Coady, Michael J; Blunck, Rikard; Lapointe, Jean-Yves

    2016-01-01

    Na-coupled cotransporters are proteins that use the trans-membrane electrochemical gradient of Na to activate the transport of a second solute. The sodium-glucose cotransporter 1 (SGLT1) constitutes a well-studied prototype of this transport mechanism but essential molecular characteristics, namely its quaternary structure and the exact arrangement of the C-terminal transmembrane segments, are still debated. After expression in Xenopus oocytes, human SGLT1 molecules (hSGLT1) were labelled on an externally accessible cysteine residue with a thiol-reactive fluorophore (tetramethylrhodamine-C5-maleimide, TMR). Addition of dipicrylamine (DPA, a negatively-charged amphiphatic fluorescence "quencher") to the fluorescently-labelled oocytes is used to quench the fluorescence originating from hSGLT1 in a voltage-dependent manner. Using this arrangement with a cysteine residue introduced at position 624 in the loop between transmembrane segments 12 and 13, the voltage-dependent fluorescence signal clearly indicated that this portion of the 12-13 loop is located on the external side of the membrane. As the 12-13 loop begins on the intracellular side of the membrane, this suggests that the 12-13 loop is re-entrant. Using fluorescence resonance energy transfer (FRET), we observed that different hSGLT1 molecules are within molecular distances from each other suggesting a multimeric complex arrangement. In agreement with this conclusion, a western blot analysis showed that hSGLT1 migrates as either a monomer or a dimer in reducing and non-reducing conditions, respectively. A systematic mutational study of endogenous cysteine residues in hSGLT1 showed that a disulfide bridge is formed between the C355 residues of two neighbouring hSGLT1 molecules. It is concluded that, 1) hSGLT1 is expressed as a disulfide bridged homodimer via C355 and that 2) a portion of the intracellular 12-13 loop is re-entrant and readily accessible from the extracellular milieu. PMID:27137918

  5. The structure of a dual-specificity tyrosine phosphorylation-regulated kinase 1A-PKC412 complex reveals disulfide-bridge formation with the anomalous catalytic loop HRD(HCD) cysteine.

    Science.gov (United States)

    Alexeeva, Marina; Åberg, Espen; Engh, Richard A; Rothweiler, Ulli

    2015-05-01

    Dual-specificity tyrosine phosphorylation-regulated kinase 1A (DYRK1A) is a protein kinase associated with neuronal development and brain physiology. The DYRK kinases are very unusual with respect to the sequence of the catalytic loop, in which the otherwise highly conserved arginine of the HRD motif is replaced by a cysteine. This replacement, along with the proximity of a potential disulfide-bridge partner from the activation segment, implies a potential for redox control of DYRK family activities. Here, the crystal structure of DYRK1A bound to PKC412 is reported, showing the formation of the disulfide bridge and associated conformational changes of the activation loop. The DYRK kinases represent emerging drug targets for several neurological diseases as well as cancer. The observation of distinct activation states may impact strategies for drug targeting. In addition, the characterization of PKC412 binding offers new insights for DYRK inhibitor discovery. PMID:25945585

  6. Carbon classified?

    DEFF Research Database (Denmark)

    Lippert, Ingmar

    2012-01-01

    . Using an actor- network theory (ANT) framework, the aim is to investigate the actors who bring together the elements needed to classify their carbon emission sources and unpack the heterogeneous relations drawn on. Based on an ethnographic study of corporate agents of ecological modernisation over...... a period of 13 months, this paper provides an exploration of three cases of enacting classification. Drawing on ANT, we problematise the silencing of a range of possible modalities of consumption facts and point to the ontological ethics involved in such performances. In a context of global warming...

  7. Diphenyl disulfide as a new bifunctional film-forming additive for high-voltage LiCoO2/graphite battery charged to 4.4 V

    Science.gov (United States)

    Zhao, Minkai; Zuo, Xiaoxi; Ma, Xiangdong; Xiao, Xin; Yu, Le; Nan, Junmin

    2016-08-01

    Diphenyl disulfide (DPDS) is evaluated as a new bifunctional electrolyte additive to improve the high-voltage performance of LiCoO2/graphite batteries. With the addition of DPDS in the electrolyte, the cell with 2.0 wt% DPDS exhibits enhanced performance in the normal voltage range of 3.0 V-4.2 V. In particular, when the cut-off potential is increased from 4.2 V to 4.4 V, the cell with 1.0 wt% DPDS also exhibits improved discharge capacity and cycle performance. Linear sweep voltammetry and cyclic voltammetry indicate that the DPDS can be reduced prior to the solvent and that the oxidative decomposition of the electrolyte can also be suppressed. In addition, X-ray diffraction, Fourier transform infrared spectroscopy and scanning electron microscopy analyses demonstrate that the solid electrolyte interface (SEI) film is produced primarily on the graphite anode via the decomposition of DPDS at normal voltage and that the SEI films induced by DPDS can be formed simultaneously on the two electrodes at higher potentials. It is hypothesized that these compact SEI films covering the electrode surface provide protection for the LiCoO2 and graphite materials and accordingly improve the cyclic performance of battery in the voltage range of 3.0 V-4.4 V.

  8. Phylogeny of the Vitamin K 2,3-Epoxide Reductase (VKOR) Family and Evolutionary Relationship to the Disulfide Bond Formation Protein B (DsbB) Family.

    Science.gov (United States)

    Bevans, Carville G; Krettler, Christoph; Reinhart, Christoph; Watzka, Matthias; Oldenburg, Johannes

    2015-07-29

    In humans and other vertebrate animals, vitamin K 2,3-epoxide reductase (VKOR) family enzymes are the gatekeepers between nutritionally acquired K vitamins and the vitamin K cycle responsible for posttranslational modifications that confer biological activity upon vitamin K-dependent proteins with crucial roles in hemostasis, bone development and homeostasis, hormonal carbohydrate regulation and fertility. We report a phylogenetic analysis of the VKOR family that identifies five major clades. Combined phylogenetic and site-specific conservation analyses point to clade-specific similarities and differences in structure and function. We discovered a single-site determinant uniquely identifying VKOR homologs belonging to human pathogenic, obligate intracellular prokaryotes and protists. Building on previous work by Sevier et al. (Protein Science 14:1630), we analyzed structural data from both VKOR and prokaryotic disulfide bond formation protein B (DsbB) families and hypothesize an ancient evolutionary relationship between the two families where one family arose from the other through a gene duplication/deletion event. This has resulted in circular permutation of primary sequence threading through the four-helical bundle protein folds of both families. This is the first report of circular permutation relating distant a-helical membrane protein sequences and folds. In conclusion, we suggest a chronology for the evolution of the five extant VKOR clades.

  9. Probing S4 and S5 segment proximity in mammalian hyperpolarization-activated HCN channels by disulfide bridging and Cd2+ coordination.

    Science.gov (United States)

    Bell, Damian C; Turbendian, Harma K; Valley, Matthew T; Zhou, Lei; Riley, John H; Siegelbaum, Steven A; Tibbs, Gareth R

    2009-06-01

    We explored the structural basis of voltage sensing in the HCN1 hyperpolarization-activated cyclic nucleotide-gated cation channel by examining the relative orientation of the voltage sensor and pore domains. The opening of channels engineered to contain single cysteine residues at the extracellular ends of the voltage-sensing S4 (V246C) and pore-forming S5 (C303) domains is inhibited by formation of disulfide or cysteine:Cd(2+) bonds. As Cd(2+) coordination is promoted by depolarization, the S4-S5 interaction occurs preferentially in the closed state. The failure of oxidation to catalyze dimer formation, as assayed by Western blotting, indicates the V246C:C303 interaction occurs within a subunit. Intriguingly, a similar interaction has been observed in depolarization-activated Shaker voltage-dependent potassium (Kv) channels at depolarized potentials but such an intrasubunit interaction is inconsistent with the X-ray crystal structure of Kv1.2, wherein S4 approaches S5 of an adjacent subunit. These findings suggest channels of opposite voltage-sensing polarity adopt a conserved S4-S5 orientation in the depolarized state that is distinct from that trapped upon crystallization.

  10. Passively Q-switched mid-infrared fluoride fiber laser around 3 µm using a tungsten disulfide (WS2) saturable absorber

    Science.gov (United States)

    Wei, Chen; Luo, Hongyu; Zhang, Han; Li, Chun; Xie, Jitao; Li, Jianfeng; Liu, Yong

    2016-10-01

    In this letter, we demonstrate a passively Q-switched Ho3+/Pr3+ co-doped fluoride fiber laser centered at 2865.7 nm using a tungsten disulfide (WS2) saturable absorber (SA) for the first time, to the best of our knowledge. A multilayer WS2 film was fabricated using the sulfidation grown method and then transferred onto an Au mirror to act as the cavity feedback and SA device in a linear cavity. Under a launched pump power of 318.5 mW, stable Q-switched pulses with an average output power of 48.4 mW were achieved with a pulse duration of 1.73 µs and repetition rate of 131.6 kHz, resulting in a pulse energy of 0.37 µJ. Our experimental results confirm that WS2 can be an effective nonlinear modulator that is suitable for pulse generation at the 3 µm waveband.

  11. The properties of mesoporous silica nanoparticles functionalized with different PEG-chain length via the disulfide bond linker and drug release in glutathione medium.

    Science.gov (United States)

    Xie, Zhifei; Gong, Huameng; Liu, Mingxing; Zhu, Hongda; Sun, Honghao

    2016-01-01

    In this paper, a novel drug-loaded material (MSNs-SS-PEG) was obtained by grafting the thiol-linked methoxy polyethylene glycol (MeOPEG-SH) onto the thiol-functionalized mesoporous silica nanoparticles (MSNs-SH) via the disulfide bond linker. In our designed experiment, three different chain lengths of PEG (PEG(1000), PEG(5000), and PEG(1000)-PEG(5000)) were used. The silica materials were characterized by Fourier transform infrared spectroscopy (FT-IR), dynamic light scattering, field emission scanning electron microscopy, transmission electron microscopy, nitrogen adsorption-desorption measurements, and X-ray diffraction. The morphology of the MSNs-SS-PEG was spherical with an average diameter of about 150 nm. Due to the covalent modification of hydrophilic MeOPEG, the MSNs-SS-PEG was coated by a thin polymer shell, showing stable and inerratic MCM-41 type mesoporous structure as well as high specific surface areas and large pore volumes. Moreover, the releases of doxorubicin hydrochloride (DOX) from these materials at 10 mM of glutathione were investigated. The PEG functionalization could effectively cap drugs in the mesoporous channels. The release of DOX from the MSNs-SS-PEG(n) revealed redox-responsive characteristic. The obtained results showed that the MSNs-SS-PEG might be promising drug delivery carrier materials, which could play an important role in the development of drug delivery. PMID:26540096

  12. Diallyl disulfide suppresses SRC/Ras/ERK signaling-mediated proliferation and metastasis in human breast cancer by up-regulating miR-34a.

    Directory of Open Access Journals (Sweden)

    Xiangsheng Xiao

    Full Text Available Diallyl disulfide (DADS is one of the major volatile components of garlic oil. DADS has various biological properties, including anticancer, antiangiogenic, and antioxidant effects. However, the anticancer mechanisms of DADS in human breast cancer have not been elucidated, particularly in vivo. In this study, we demonstrated that the expression of miR-34a was up-regulated in DADS-treated MDA-MB-231 cells. miR-34a not only inhibited breast cancer growth but also enhanced the antitumor effect of DADS, both in vitro and in vivo. Furthermore, Src was identified as a target of miR-34a, with miR-34a inhibiting SRC expression and consequently triggering the suppression of the SRC/Ras/ERK pathway. These results suggest that DADS could be a promising anticancer agent for breast cancer. miR-34a may also demonstrate a potential gene therapy agent that could enhance the antitumor effects of DADS.

  13. The role of reactive oxygen species (ROS) production on diallyl disulfide (DADS) induced apoptosis and cell cycle arrest in human A549 lung carcinoma cells

    Energy Technology Data Exchange (ETDEWEB)

    Wu Xinjiang [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany); Kassie, Fekadu [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany); Mersch-Sundermann, Volker [Institute of Indoor and Environmental Toxicology, Faculty of Medicine, Justus-Liebig-University of Giessen, Aulweg 123, D-35385 Giessen (Germany)]. E-mail: Volker.mersch-sundermann@uniklinikum-giessen.de

    2005-11-11

    Diallyl disulfide (DADS), an oil soluble constituent of garlic (Allium sativum), has been reported to cause antimutagentic and anticarcinogenic effects in vitro and in vivo by modulating phases I and II enzyme activities. In recent years, several studies suggested that the chemopreventive effects of DADS can also be attributed to induction of cell cycle arrest and apoptosis in cancer cells. In the present study, we reported that DADS-induced cell cycle arrest at G2/M and apoptosis in human A549 lung cancer cells in a time- and dose-dependent manner. Additionally, a significant increase of intracellular reactive oxygen species (ROS) was induced in A549 cells less than 0.5 h after DADS treatment, indicating that ROS may be an early event in DADS-modulated apoptosis. Treatment of A549 cells with N-acetyl cysteine (NAC) completely abrogated DADS-induced cell cycle arrest and apoptosis. The result indicated that oxidative stress modulates cell proliferation and cell death induced by DADS.

  14. Enhancement of lipase r27RCL production in Pichia pastoris by regulating gene dosage and co-expression with chaperone protein disulfide isomerase.

    Science.gov (United States)

    Sha, Chong; Yu, Xiao-Wei; Lin, Nai-Xin; Zhang, Meng; Xu, Yan

    2013-12-10

    Pichia pastoris has been successfully used in the production of many secreted and intracellular recombinant proteins, but there is still a large room of improvement for this expression system. Two factors drastically influence the lipase r27RCL production from Rhizopus chinensis CCTCC M201021, which are gene dosage and protein folding in the endoplasmic reticulum (ER). Regarding the effect of gene dosage, the enzyme activity for recombinant strain with three copies lipase gene was 1.95-fold higher than that for recombinant strain with only one copy lipase gene. In addition, the lipase production was further improved by co-expression with chaperone PDI involved in the disulfide bond formation in the ER. Overall, the maximum enzyme activity reached 355U/mL by the recombinant strain with one copy chaperone gene PDI plus five copies lipase gene proRCL in shaking flasks, which was 2.74-fold higher than that for the control strain with only one copy lipase gene. Overall, co-expression with PDI vastly increased the capacity for processing proteins of ER in P. pastoris. PMID:24315648

  15. Redox-Triggered Release of Moxifloxacin from Mesoporous Silica Nanoparticles Functionalized with Disulfide Snap-Tops Enhances Efficacy Against Pneumonic Tularemia in Mice.

    Science.gov (United States)

    Lee, Bai-Yu; Li, Zilu; Clemens, Daniel L; Dillon, Barbara Jane; Hwang, Angela A; Zink, Jeffrey I; Horwitz, Marcus A

    2016-07-01

    Effective and rapid treatment of tularemia is needed to reduce morbidity and mortality of this potentially fatal infectious disease. The etiologic agent, Francisella tularensis, is a facultative intracellular bacterial pathogen which infects and multiplies to high numbers in macrophages. Nanotherapeutics are particularly promising for treatment of infectious diseases caused by intracellular pathogens, whose primary host cells are macrophages, because nanoparticles preferentially target and are avidly internalized by macrophages. A mesoporous silica nanoparticle (MSN) has been developed functionalized with disulfide snap-tops that has high drug loading and selectively releases drug intracellularly in response to the redox potential. These nanoparticles, when loaded with Hoechst fluorescent dye, release their cargo exclusively intracellularly and stain the nuclei of macrophages. The MSNs loaded with moxifloxacin kill F. tularensis in macrophages in a dose-dependent fashion. In a mouse model of lethal pneumonic tularemia, MSNs loaded with moxifloxacin prevent weight loss, illness, and death, markedly reduce the burden of F. tularensis in the lung, liver, and spleen, and are significantly more efficacious than an equivalent amount of free drug. An important proof-of-principle for the potential therapeutic use of a novel nanoparticle drug delivery platform for the treatment of infectious diseases is provided. PMID:27246117

  16. Ytterbium-doped fiber laser passively mode locked by few-layer Molybdenum Disulfide (MoS2) saturable absorber functioned with evanescent field interaction.

    Science.gov (United States)

    Du, Juan; Wang, Qingkai; Jiang, Guobao; Xu, Changwen; Zhao, Chujun; Xiang, Yuanjiang; Chen, Yu; Wen, Shuangchun; Zhang, Han

    2014-01-01

    By coupling few-layer Molybdenum Disulfide (MoS2) with fiber-taper evanescent light field, a new type of MoS2 based nonlinear optical modulating element had been successfully fabricated as a two-dimensional layered saturable absorber with strong light-matter interaction. This MoS2-taper-fiber device is not only capable of passively mode-locking an all-normal-dispersion ytterbium-doped fiber laser and enduring high power laser excitation (up to 1 W), but also functions as a polarization sensitive optical modulating component (that is, different polarized light can induce different nonlinear optical response). Thanks to the combined advantages from the strong nonlinear optical response in MoS2 together with the sufficiently-long-range interaction between light and MoS2, this device allows for the generation of high power stable dissipative solitons at 1042.6 nm with pulse duration of 656 ps and a repetition rate of 6.74 MHz at a pump power of 210 mW. Our work may also constitute the first example of MoS2-enabled wave-guiding photonic device, and potential y give some new insights into two-dimensional layered materials related photonics. PMID:25213108

  17. Protein disulfide isomerase-like protein 1-1 controls endosperm development through regulation of the amount and composition of seed proteins in rice.

    Directory of Open Access Journals (Sweden)

    Yeon Jeong Kim

    Full Text Available Protein disulfide isomerase (PDI is a chaperone protein involved in oxidative protein folding by acting as a catalyst and assisting folding in the endoplasmic reticulum (ER. A genome database search showed that rice contains 19 PDI-like genes. However, their functions are not clearly identified. This paper shows possible functions of rice PDI-like protein 1-1 (PDIL1-1 during seed development. Seeds of the T-DNA insertion PDIL1-1 mutant, PDIL1-1Δ, identified by genomic DNA PCR and western blot analysis, display a chalky phenotype and a thick aleurone layer. Protein content per seed was significantly lower and free sugar content higher in PDIL1-1Δ mutant seeds than in the wild type. Proteomic analysis of PDIL1-1Δ mutant seeds showed that PDIL1-1 is post-translationally regulated, and its loss causes accumulation of many types of seed proteins including glucose/starch metabolism- and ROS (reactive oxygen species scavenging-related proteins. In addition, PDIL1-1 strongly interacts with the cysteine protease OsCP1. Our data indicate that the opaque phenotype of PDIL1-1Δ mutant seeds results from production of irregular starch granules and protein body through loss of regulatory activity for various proteins involved in the synthesis of seed components.

  18. Variability in automated assignment of NOESY spectra and three-dimensional structure determination: A test case on three small disulfide-bonded proteins

    International Nuclear Information System (INIS)

    Three independent runs of automatic assignment and structure calculations were performed on three small proteins, calcicludine from the venom of the green mamba Dendroaspis angusticeps, κ-conotoxin PVIIA from the purple cone Conus purpurascens and HsTX1, a short scorpion toxin from the venom of Heterometrus spinnifer. At the end of all the runs, the number of cross peaks which remained unassigned (0.6%, 1.4% and 2% for calcicludine, κ-conotoxin and HsTX1, respectively), as well as the number of constraints which were rejected as producing systematic violations (2.7%, 1.0%, and 1.4% for calcicludine, κ-conotoxin and HsTX1, respectively) were low. The conformation of the initial model used in the procedure (linear model or constructed by homology) has no influence on the final structures. Mainly two parameters control the procedure: the chemical shift tolerance and the cut-off distance. Independent runs of structure calculations, using the same parameters, yield structures for which the rmsd between averaged structures and the rmsd around each averaged structure were of the same order of magnitude. A different cut-off distance and a different chemical shift tolerance yield rmsd values on final average structures which did not differ more than 0.5 A compared to the rmsd obtained around the averaged structure for each calculation. These results show that the procedure is robust when applied to such a small disulfide-bonded protein

  19. The role of reactive oxygen species (ROS) production on diallyl disulfide (DADS) induced apoptosis and cell cycle arrest in human A549 lung carcinoma cells

    International Nuclear Information System (INIS)

    Diallyl disulfide (DADS), an oil soluble constituent of garlic (Allium sativum), has been reported to cause antimutagentic and anticarcinogenic effects in vitro and in vivo by modulating phases I and II enzyme activities. In recent years, several studies suggested that the chemopreventive effects of DADS can also be attributed to induction of cell cycle arrest and apoptosis in cancer cells. In the present study, we reported that DADS-induced cell cycle arrest at G2/M and apoptosis in human A549 lung cancer cells in a time- and dose-dependent manner. Additionally, a significant increase of intracellular reactive oxygen species (ROS) was induced in A549 cells less than 0.5 h after DADS treatment, indicating that ROS may be an early event in DADS-modulated apoptosis. Treatment of A549 cells with N-acetyl cysteine (NAC) completely abrogated DADS-induced cell cycle arrest and apoptosis. The result indicated that oxidative stress modulates cell proliferation and cell death induced by DADS

  20. Carbon Nanomembranes.

    Science.gov (United States)

    Turchanin, Andrey; Gölzhäuser, Armin

    2016-08-01

    Carbon nanomembranes (CNMs) are synthetic 2D carbon sheets with tailored physical or chemical properties. These depend on the structure, molecular composition, and surroundings on either side. Due to their molecular thickness, they can be regarded as "interfaces without bulk" separating regions of different gaseous, liquid, or solid components and controlling the materials exchange between them. Here, a universal scheme for the fabrication of 1 nm-thick, mechanically stable, functional CNMs is presented. CNMs can be further modified, for example perforated by ion bombardment or chemically functionalized by the binding of other molecules onto the surfaces. The underlying physical and chemical mechanisms are described, and examples are presented for the engineering of complex surface architectures, e.g., nanopatterns of proteins, fluorescent dyes, or polymer brushes. A simple transfer procedure allows CNMs to be placed on various support structures, which makes them available for diverse applications: supports for electron and X-ray microscopy, nanolithography, nanosieves, Janus nanomembranes, polymer carpets, complex layered structures, functionalization of graphene, novel nanoelectronic and nanomechanical devices. To close, the potential of CNMs in filtration and sensorics is discussed. Based on tests for the separation of gas molecules, it is argued that ballistic membranes may play a prominent role in future efforts of materials separation. PMID:27281234

  1. Trading forest carbon

    Science.gov (United States)

    The nature of carbon in forests is discussed from the perspective of carbon trading. Carbon inventories, specifically in the area of land use and forestry are reviewed for the Pacific Northwest. Carbon turnover in forests is discussed as it relates to carbon sequestration. Scient...

  2. Carbon Farming as a Carbon Negative Technology

    Science.gov (United States)

    Anderson, C.; Laird, D.; Hayes, D. J.

    2015-12-01

    Carbon farms have a pivotal role in national and international efforts to mitigate and adapt to climate change. A carbon farm in its broadest sense is one that reduces greenhouse gas (GHG) emissions or captures and holds carbon in vegetation and soils. Their capacity to remove carbon from the air and store it safely and permanently, while providing additional human and ecosystem benefits, means they could contribute significantly to national efforts to stabilize or reduce GHGs. We examine carbon farms in the context of corn and soybean production agriculture. We illustrate, using Iowa data but with relevance across United States corn and soybean production, the potential for carbon farms to reduce human GHG emissions and sequester carbon permanently at a rate that has meaningful impact on global greenhouse gas concentration. Carbon has been viewed as a next generation cash crop in Iowa for over a decade. The carbon farm perspective, however, goes beyond carbon as cash crop to make carbon the center of an entire farm enterprise. The transformation is possible through slight adjustment crop practices mixed with advances in technology to sequester carbon through biochar. We examine carbon balance of Iowa agriculture given only the combination of slight reduction in fertilizer and sequestration by biochar. We find the following. Iowa carbon farms could turn Iowa agriculture into a carbon sink. The estimated range of GHG reduction by statewide implementation of carbon farms is 19.46 to 90.27 MMt CO2-equivalent (CO2-e), while the current agricultural CO2-e emission estimate is 35.38 MMt CO2-e. Iowa carbon farm GHG reduction would exceed Iowa GHG reduction by wind energy (8.7 MMt CO2-e) and could exceed combined reductions from wind energy and corn grain ethanol (10.7 MMt CO2-e; 19.4 MMt CO2-e combined). In fact, Iowa carbon farms alone could exceed GHG reduction from national corn grain ethanol production (39.6 MMt CO2-e). A carbon price accessible to agricultural

  3. Net Ecosystem Carbon Flux

    Data.gov (United States)

    U.S. Geological Survey, Department of the Interior — Net Ecosystem Carbon Flux is defined as the year-over-year change in Total Ecosystem Carbon Stock, or the net rate of carbon exchange between an ecosystem and the...

  4. Carbon Monoxide (CO)

    Science.gov (United States)

    ... IAQ) » Carbon Monoxide's Impact on Indoor Air Quality Carbon Monoxide's Impact on Indoor Air Quality On this ... length of exposure. Top of Page Sources of Carbon Monoxide Sources of CO include: unvented kerosene and ...

  5. Carbon Monoxide Poisoning

    Science.gov (United States)

    ... Recommend on Facebook Tweet Share Compartir What is Carbon Monoxide? Carbon monoxide, or “CO,” is an odorless, colorless gas that can kill you. Carbon monoxide detector Where is CO found? CO is ...

  6. Integral Ring Carbon-Carbon Piston

    Science.gov (United States)

    Northam, G. Burton (Inventor)

    1999-01-01

    An improved structure for a reciprocating internal combustion engine or compressor piston fabricate from carbon-carbon composite materials is disclosed. An integral ring carbon-carbon composite piston, disclosed herein, reduces the need for piston rings and for small clearances by providing a small flexible, integral component around the piston that allows for variation in clearance due to manufacturing tolerances, distortion due to pressure and thermal loads, and variations in thermal expansion differences between the piston and cylinder liner.

  7. Carbon dioxide sequestration by mineral carbonation

    OpenAIRE

    Huijgen, W.J.J.

    2007-01-01

    The increasing atmospheric carbon dioxide (CO2) concentration, mainly caused by fossil fuel combustion, has lead to concerns about global warming. A possible technology that can contribute to the reduction of carbon dioxide emissions is CO2 sequestration by mineral carbonation. The basic concept behind mineral CO2 sequestration is the mimicking of natural weathering processes in which calcium or magnesium containing minerals react with gaseous CO2 and form solid calcium or magnesium carbonate...

  8. Effects of dimethyl disulfide on microbial communities in protectorate soils under continuous cropping%二甲基二硫熏蒸对保护地连作土壤微生物群落的影响

    Institute of Scientific and Technical Information of China (English)

    王方艳; 王秋霞; 颜冬冬; 毛连纲; 郭美霞; 燕平梅; 曹坳程

    2011-01-01

    -borne diseases. As broad-spectrum agents, however, fiimigants also have side effects on non-target organisms. Dimethyl disulfide (DMDS) is a new alternative to methyl bromide (MeBr) that reduces plant fungal pathogens and nematodes. DMDS is therefore recommended by the Methyl Bromide Technical Options Committee of the United Nations Environment Program (UNEP). This study was an attempt to identify the effects of DMDS on microbial communities in protectorate soils under continuous cropping. The efficacy of DMDS was evaluated by bio-assay in the laboratory. The efficacy of DMDS on the Fusarium spp. And Phytophthora spp. Was observed after fumigations. The LCS0 of DMDS with different concentrations (170.00 mgkg-1, 85.20 mg-kg-1, 42.50 mg-kg-1, 21.30 mg-kg-1 and 10.62 mg-kg-1) was 42.08 mgkg-1 and 115.15 mg-kg1, respectively to Fusarium spp. And Phytophthora spp. Microbial community structures after DMDS fumigation were evaluated using BIOLOG Ecoplates under laboratory conditions. Compared with the un-treated/control plants, the average well color development (AWCD) of the DMDS 10.62 mg-kgf1, 42.50 mg-kg"1 and 170.00 mgkg1 respectively increased by 8.46%, 6.02% and 19.31%, 0 day after fumigation and 120 h after sample incubation. AWCD increased by 1.87%, 3.47% and 8.01%, respectively, 240 h after incubation; which indicated that DMDS promoted the growth of microbes. AWCD of treated samples were close to the control at 14 days after fumigation. The indices of Shannon and Simpson at 0 day after fumigation were higher than lhat of the control, recovering to the levels of the control 7 days after fumigation. Based on Mclntosh index, there was no significant difference between the fumigation treatments and the control. Principal component analysis of substrate reaction reflected that the use of carbon sources by microbial community was obviously different in the treatments immediately after fumigation. It was, however, close to the control 14 days after fumigation. The study showed that

  9. Carbon Residence Times in Pedogenic Carbonate Pools

    Science.gov (United States)

    Monger, H.; Feng, Y.; Karnjanapiboonwang, A.

    2013-12-01

    Soil carbonate is a huge pool of terrestrial carbon that contains at least 930 to 940 Pg C and has influx rates on the order of 1 to 12 g CaCO3/m2/yr. Such large mass to flux ratios yield long mean residence times for carbon (e.g., 85,000 years)--assuming steady state. Like other global carbon pools, the soil carbonate pool has smaller sub-pools with higher influx rates and shorter mean residence times. For example, pedogenic carbonate in coppice dunes known to have formed since 1858 and carbonate formed on lithic artifacts in soils at archaeology sites suggests mean residence times can be as short as 120 years--again assuming steady state. Harder to assess are efflux rates as CO2 emissions or bicarbonate leaching. Some Bowen-ratio studies have nevertheless found evidence for CO2 emissions resulting from carbonate dissolution, and other studies have found evidence for bicarbonate leaching based on dissolution pipes through calcic horizons using soil morphology studies. Since an understanding of mean residence times are prerequisite for a better understanding of soil carbonate in the global carbon cycle, especially in a scenario of an expanding Aridosphere, more influx and efflux measurements are needed to evaluate the possibility of carbon sequestration by soil carbonate in hyperarid, arid, semiarid, or subhumid soils.

  10. Carbon dioxide sequestration by mineral carbonation

    NARCIS (Netherlands)

    Huijgen, W.J.J.

    2007-01-01

    The increasing atmospheric carbon dioxide (CO2) concentration, mainly caused by fossil fuel combustion, has lead to concerns about global warming. A possible technology that can contribute to the reduction of carbon dioxide emissions is CO2 sequestration by mineral carbonation. The basic concept beh

  11. Diallyl Disulfide (DADS), a Constituent of Garlic, Inactivates NF-κB and Prevents Colitis-Induced Colorectal Cancer by Inhibiting GSK-3β.

    Science.gov (United States)

    Saud, Shakir M; Li, Weidong; Gray, Zane; Matter, Matthias S; Colburn, Nancy H; Young, Matthew R; Kim, Young S

    2016-07-01

    There is a strong belief that garlic has medicinal properties and may even reduce the risk of developing certain cancers including those of the gastrointestinal tract. The chemopreventive effects of garlic may be attributed to the anti-inflammatory properties of the sulfur-containing constituents of garlic, which includes diallyl disulfide (DADS). Here, we demonstrate that DADS prevented colorectal tumorigenesis in a mouse model of colitis-induced colorectal cancer. Supplementation with 85 ppm of DADS (60 mg daily human equivalent dose) in the diet of FVB/N mice treated with chemical carcinogen azoxymethane (AOM) and colonic irritant dextran sodium sulfate (DSS) resulted in the reduction in tumor incidence, tumor number, and tumor burden by 21.54%, 47.3%, and 66.4%, respectively. Further analysis revealed that mice fed the DADS-supplemented diet resolved the initial DSS-induced inflammation faster than those on the control diet, preventing prolonged inflammation and cellular transformation. Subsequent mechanistic studies in vitro suggest that DADS chemopreventive effects are mediated through NF-κB signaling. When SW480 colorectal cancer cells were treated with DADS, NF-κB nuclear localization and activity were diminished. Interestingly, NF-κB suppression was found to be dependent on DADS inhibition of GSK-3β, a positive regulator of NF-κB. Inhibition of GSK-3β and loss of nuclear NF-κB activity were also observed in vivo in AOM/DSS-treated mice fed a diet supplemented with 85 ppm DADS. Our results indicate that DADS can prevent tumorigenesis by suppressing inflammation, a process largely involving GSK-3β inhibition and consequential reduction in NF-κB nuclear localization. Cancer Prev Res; 9(7); 607-15. ©2016 AACR. PMID:27138790

  12. Folic acid-targeted disulfide-based cross-linking micelle for enhanced drug encapsulation stability and site-specific drug delivery against tumors.

    Science.gov (United States)

    Zhang, Yumin; Zhou, Junhui; Yang, Cuihong; Wang, Weiwei; Chu, Liping; Huang, Fan; Liu, Qiang; Deng, Liandong; Kong, Deling; Liu, Jianfeng; Liu, Jinjian

    2016-01-01

    Although the shortcomings of small molecular antitumor drugs were efficiently improved by being entrapped into nanosized vehicles, premature drug release and insufficient tumor targeting demand innovative approaches that boost the stability and tumor responsiveness of drug-loaded nanocarriers. Here, we show the use of the core cross-linking method to generate a micelle with enhanced drug encapsulation ability and sensitivity of drug release in tumor. This kind of micelle could increase curcumin (Cur) delivery to HeLa cells in vitro and improve tumor accumulation in vivo. We designed and synthesized the core cross-linked micelle (CCM) with polyethylene glycol and folic acid-polyethylene glycol as the hydrophilic units, pyridyldisulfide as the cross-linkable and hydrophobic unit, and disulfide bond as the cross-linker. CCM showed spherical shape with a diameter of 91.2 nm by the characterization of dynamic light scattering and transmission electron microscope. Attributed to the core cross-linking, drug-loaded CCM displayed higher Nile Red or Cur-encapsulated stability and better sensitivity to glutathione than noncross-linked micelle (NCM). Cellular uptake and in vitro antitumor studies proved the enhanced endocytosis and better cytotoxicity of CCM-Cur against HeLa cells, which had a high level of glutathione. Meanwhile, the folate receptor-mediated drug delivery (FA-CCM-Cur) further enhanced the endocytosis and cytotoxicity. Ex vivo imaging studies showed that CCM-Cur and FA-CCM-Cur possessed higher tumor accumulation until 24 hours after injection. Concretely, FA-CCM-Cur exhibited the highest tumor accumulation with 1.7-fold of noncross-linked micelle Cur and 2.8-fold of free Cur. By combining cross-linking of the core with active tumor targeting of FA, we demonstrated a new and effective way to design nanocarriers for enhanced drug encapsulation, smart tumor responsiveness, and elevated tumor accumulation. PMID:27051287

  13. Enolase 1 (ENO1 and protein disulfide-isomerase associated 3 (PDIA3 regulate Wnt/β-catenin-driven trans-differentiation of murine alveolar epithelial cells

    Directory of Open Access Journals (Sweden)

    Kathrin Mutze

    2015-08-01

    Full Text Available The alveolar epithelium represents a major site of tissue destruction during lung injury. It consists of alveolar epithelial type I (ATI and type II (ATII cells. ATII cells are capable of self-renewal and exert progenitor function for ATI cells upon alveolar epithelial injury. Cell differentiation pathways enabling this plasticity and allowing for proper repair, however, are poorly understood. Here, we applied proteomics, expression analysis and functional studies in primary murine ATII cells to identify proteins and molecular mechanisms involved in alveolar epithelial plasticity. Mass spectrometry of cultured ATII cells revealed a reduction of carbonyl reductase 2 (CBR2 and an increase in enolase 1 (ENO1 and protein disulfide-isomerase associated 3 (PDIA3 protein expression during ATII-to-ATI cell trans-differentiation. This was accompanied by increased Wnt/β-catenin signaling, as analyzed by qRT-PCR and immunoblotting. Notably, ENO1 and PDIA3, along with T1α (podoplanin; an ATI cell marker, exhibited decreased protein expression upon pharmacological and molecular Wnt/β-catenin inhibition in cultured ATII cells, whereas CBR2 levels were stabilized. Moreover, we analyzed primary ATII cells from mice with bleomycin-induced lung injury, a model exhibiting activated Wnt/β-catenin signaling in vivo. We observed reduced CBR2 significantly correlating with surfactant protein C (SFTPC, whereas ENO1 and PDIA3 along with T1α were increased in injured ATII cells. Finally, siRNA-mediated knockdown of ENO1, as well as PDIA3, in primary ATII cells led to reduced T1α expression, indicating diminished cell trans-differentiation. Our data thus identified proteins involved in ATII-to-ATI cell trans-differentiation and suggest a Wnt/β-catenin-driven functional role of ENO1 and PDIA3 in alveolar epithelial cell plasticity in lung injury and repair.

  14. Enhancing sensitivity and selectivity in a label-free colorimetric sensor for detection of iron(II) ions with luminescent molybdenum disulfide nanosheet-based peroxidase mimetics.

    Science.gov (United States)

    Wang, Yong; Hu, Jie; Zhuang, Qianfen; Ni, Yongnian

    2016-06-15

    In the present study, we demonstrated that the luminescent molybdenum disulfide (MoS2) nanosheets, which were prepared hydrothermally by using sodium molybdate and thiourea as precursors, possessed peroxidase-like activity, and could catalyze the oxidation of peroxidase substrate o-phenylenediamine (OPD) in the presence of hydrogen peroxide (H2O2) to produce a yellow color reaction. Further addition of Fe(2+) into the nanosheets led to peroxidase mimetics with greatly enhanced catalytic activity. The observation was exploited to develop a label-free colorimetric nanozyme sensor for detection of Fe(2+). The fabricated MoS2/OPD/H2O2 sensor showed a wide linear range of 0.01-0.8 µM with a detection limit of 7 nM. Moreover, it was found that the MoS2/OPD/H2O2 sensor displayed enhanced sensitivity and selectivity toward Fe(2+) compared with the OPD/H2O2 sensor, suggesting that the MoS2 nanosheets could improve the performance of the Fe(2+) sensor. An advanced chemometrics algorithm, multivariate curve resolution by alternating least squares (MCR-ALS), was further applied to interpret the origin of enhancing sensitivity and selectivity in the Fe(2+) sensor with the MoS2 nanosheets. The time-dependent UV-vis spectral data of the studied systems were collected, and submitted to the MCR-ALS. The results showed that the increased sensitivity and selectivity of the MoS2/OPD/H2O2 sensor for Fe(2+) detection likely arose from its large reaction rate constant. Finally, the proposed MoS2/OPD/H2O2 sensor was successfully applied for detection of Fe(2+) in water samples.

  15. Pyridine Nucleotide Complexes with Bacillus anthracis Coenzyme A-Disulfide Reductase: A Structural Analysis of Dual NAD(P)H Specificity

    Energy Technology Data Exchange (ETDEWEB)

    Wallen,J.; Paige, C.; Mallett, T.; Karplus, P.; Claiborne, A.

    2008-01-01

    We have recently reported that CoASH is the major low-molecular weight thiol in Bacillus anthracis, and we have now characterized the kinetic and redox properties of the B. anthracis coenzyme A-disulfide reductase (CoADR, BACoADR) and determined the crystal structure at 2.30 Angstroms resolution. While the Staphylococcus aureus and Borrelia burgdorferi CoADRs exhibit strong preferences for NADPH and NADH, respectively, B. anthracis CoADR can use either pyridine nucleotide equally well. Sequence elements within the respective NAD(P)H-binding motifs correctly reflect the preferences for S. aureus and Bo. burgdorferi CoADRs, but leave questions as to how BACoADR can interact with both pyridine nucleotides. The structures of the NADH and NADPH complexes at ca. 2.3 Angstroms resolution reveal that a loop consisting of residues Glu180-Thr187 becomes ordered and changes conformation on NAD(P)H binding. NADH and NADPH interact with nearly identical conformations of this loop; the latter interaction, however, involves a novel binding mode in which the 2'-phosphate of NADPH points out toward solvent. In addition, the NAD(P)H-reduced BACoADR structures provide the first view of the reduced form (Cys42-SH/CoASH) of the Cys42-SSCoA redox center. The Cys42-SH side chain adopts a new conformation in which the conserved Tyr367'-OH and Tyr425'-OH interact with the nascent thiol(ate) on the flavin si-face. Kinetic data with Y367F, Y425F, and Y367, 425F BACoADR mutants indicate that Tyr425' is the primary proton donor in catalysis, with Tyr367' functioning as a cryptic alternate donor in the absence of Tyr425'.

  16. Tungsten disulfide nanosheet and exonuclease III co-assisted amplification strategy for highly sensitive fluorescence polarization detection of DNA glycosylase activity

    International Nuclear Information System (INIS)

    Herein, we introduced a tungsten disulfide (WS2) nanosheet and exonuclease III (Exo III) co-assisted signal amplification strategy for highly sensitive fluorescent polarization (FP) assay of DNA glycosylase activity. Two DNA glycosylases, uracil-DNA glycosylase (UDG) and human 8-oxoG DNA glycosylase 1 (hOGG1), were tested. A hairpin-structured probe (HP) which contained damaged bases in the stem was used as the substrate. The removal of damaged bases from substrate by DNA glycosylase would lower the melting temperature of HP. The HP was then opened and hybridized with a FAM dye-labeled single strand DNA (DP), generating a duplex with a recessed 3′-terminal of DP. This design facilitated the Exo III-assisted amplification by repeating the hybridization and digestion of DP, liberating numerous FAM fluorophores which could not be adsorbed on WS2 nanosheet. Thus, the final system exhibited a small FP signal. However, in the absence of DNA glycosylases, no hybridization between DP and HP was occurred, hampering the hydrolysis of DP by Exo III. The intact DP was then adsorbed on the surface of WS2 nanosheet that greatly amplified the mass of the labeled-FAM fluorophore, resulting in a large FP value. With the co-assisted amplification strategy, the sensitivity was substantially improved. In addition, this method was applied to detect UDG activity in cell extracts. The study of the inhibition of UDG was also performed. Furthermore, this method is simple in design, easy in implementation, and selective, which holds potential applications in the DNA glycosylase related mechanism research and molecular diagnostics. - Highlights: • A fluorescence polarization strategy for DNA glycosylase activity detection was developed. • The present method was based on WS2 nanosheet and exonuclease III co-assisted signal amplification. • A high sensitivity and desirable selectivity were achieved. • This method provides a promising universal platform for DNA glycosylase activity

  17. The Two-Dimensional Nanocomposite of Molybdenum Disulfide and Nitrogen-Doped Graphene Oxide for Efficient Counter Electrode of Dye-Sensitized Solar Cells.

    Science.gov (United States)

    Cheng, Chao-Kuang; Lin, Che-Hsien; Wu, Hsuan-Chung; Ma, Chen-Chi M; Yeh, Tsung-Kuang; Chou, Huei-Yu; Tsai, Chuen-Horng; Hsieh, Chien-Kuo

    2016-12-01

    In this study, we reported the synthesis of the two-dimensional (2D) nanocomposite of molybdenum disulfide and nitrogen-doped graphene oxide (MoS2/nGO) as a platinum-free counter electrode (CE) for dye-sensitized solar cells (DSSCs). X-ray photoelectron spectroscopy (XPS), high-resolution transmission electron microscopy (HRTEM), and Raman spectroscopy were used to examine the characteristics of the 2D nanocomposite of MoS2/nGO. The cyclic voltammetry (CV), electrochemical impedance spectra (EIS), and the Tafel polarization measurements were carried out to examine the electrocatalytic abilities. XPS and Raman results showed the 2D behaviors of the prepared nanomaterials. HRTEM micrographs showed the direct evidence of the 2D nanocomposite of MoS2/nGO. The results of electrocatalytic examinations indicated the MoS2/nGO owning the low charge transfer resistance, high electrocatalytic activity, and fast reaction kinetics for the reduction of triiodide to iodide on the electrolyte-electrode interface. The 2D nanocomposite of MoS2/nGO combined the advantages of the high specific surface of nGO and the plenty edge sites of MoS2 and showed the promoted properties different from those of their individual constituents to create a new outstanding property. The DSSC with MoS2/nGO nanocomposite CE showed a photovoltaic conversion efficiency (PCE) of 5.95 % under an illumination of AM 1.5 (100 mW/cm(2)), which was up to 92.2 % of the DSSC with the conventional platinum (Pt) CE (PCE = 6.43 %). These results reveal the potential of the MoS2/nGO nanocomposite in the use of low-cost, scalable, and efficient Pt-free CEs for DSSCs. PMID:26925865

  18. Apoptosis Induction of Human Prostate Carcinoma DU145 Cells by Diallyl Disulfide via Modulation of JNK and PI3K/AKT Signaling Pathways

    Directory of Open Access Journals (Sweden)

    Young Hyun Yoo

    2012-11-01

    Full Text Available Diallyl disulfide (DADS, a sulfur compound derived from garlic, has various biological properties, such as anticancer, antiangiogenic and anti-inflammatory effects. However, the mechanisms of action underlying the compound's anticancer activity have not been fully elucidated. In this study, the apoptotic effects of DADS were investigated in DU145 human prostate carcinoma cells. Our results showed that DADS markedly inhibited the growth of the DU145 cells by induction of apoptosis. Apoptosis was accompanied by modulation of Bcl-2 and inhibitor of apoptosis protein (IAP family proteins, depolarization of the mitochondrial membrane potential (MMP, ΔΨm and proteolytic activation of caspases. We also found that the expression of death-receptor 4 (DR4 and Fas ligand (FasL proteins was increased and that the level of intact Bid proteins was down-regulated by DADS. Moreover, treatment with DADS induced phosphorylation of mitogen-activated protein kinases (MAPKs, including extracellular-signal regulating kinase (ERK, p38 MAPK and c-Jun N-terminal kinase (JNK. A specific JNK inhibitor, SP600125, significantly blocked DADS-induced-apoptosis, whereas inhibitors of the ERK (PD98059 and p38 MAPK (SB203580 had no effect. The induction of apoptosis was also accompanied by inactivation of phosphatidylinositol 3-kinase (PI3K/Akt and the PI3K inhibitor LY29004 significantly increased DADS-induced cell death. These findings provide evidence demonstrating that the proapoptotic effect of DADS is mediated through the activation of JNK and the inhibition of the PI3K/Akt signaling pathway in DU145 cells.

  19. Diallyl disulfide suppresses growth of HL-60 cell through increasing his-tone acetylation and p21WAF1 expression in vivo and in vitro

    Institute of Scientific and Technical Information of China (English)

    Jie ZHAO; Wei-guo HUANG; Jie HE; Hui TAN; Qian-jin LIAO; Qi SU

    2006-01-01

    Aim: To examine the differentiation induction and growth inhibition of HL-60 cells by diallyl disulfide (DADS), and its relationship with the alterations of histone acetylation and p21WAF1 expression in vitro and in vivo. Methods: Differentiation was studied by nitroblue tetrazolium (NET) reduction of HL-60 cell in vitro. HL-60 cells 5xl06 were injected into the right side of the peritoneal cavity of severe combined immunodeficiency (SCID) mice. When the peritoneal neoplasms were detected, the SCID mice were randomly divided into 3 groups and received an ip injection of vehicle alone (NS), DADS or sodium butyrate (SB). The growth inhibition of peritoneal neoplasms induced by DADS was observed by a growth curve. The cycle distribution of HL-60 cells in SCID mice was monitored by flow cytometry. The expression of acetylated histone H3, H4 and p21WAF1 were measured by Western blot. Results: After treatment with DADS for 0-72 h, the NET reduction ability of HL-60 cells increased in a time-dependent manner, compared with no treatment of HL-60 cells. In the HL-60 cells treated with DADS for 24 h, the expression of acetylated histone H3, H4, and p21WAF1 increased obviously. After treatment with DADS, tumor growth was markedly suppressed. HL-60 cells from mice treated with DADS were blocked in the G1 phase, from 25.4% to 63.4%. The tumors from the mice treated with DADS showed an increase of acetylated histone H3, H4, and p21WAF1. Conclusion: DADS could induce differentiation and inhibit the growth of HL-60 cells through increasing the expression of acetylated histone H3, H4, and p21WAF1 in vitro and in vivo.

  20. Synergistic effect of carbon monoxide with other biologically active injurious factors on the organism

    Energy Technology Data Exchange (ETDEWEB)

    Pankow, D.; Ponsold, W.

    1974-09-01

    The combined effects on biological organisms are reported for carbon monoxide and carbon dioxide, nitrogen oxides, sodium nitrite, hydrocyanic acid, carbon disulfide, sulfur dioxide, ammonia, hydrogen peroxide, ethanol, trichloroethylene, carbon tetrachloride, methane, benzene, iodine acetate, cholesterol, benzpyrene, hexobarbitol, zoxazolamine, nembutal, luminal, morphine, adrenalin, persantin, cytochrome c, aldrin, carbaryl, cyclodiene epoxide; and physical influences such as ambient temperature, atmospheric pressure, ionizing radiation, noise, and vibration. A literature review shows that with increasing CO/sub 2/ and decreasing oxygen concentration in the inhalation air, the toxicity of CO increased in experiments with mice and canaries. Oxides of nitrogen enhance the toxic effect of CO in an additive way and at times synergistically. At 500 m from a metallurgical plant in the USSR the maximum allowable immission concentrations for CO and SO/sub 2/ were exceeded. In children residing there a higher normal erythrocyte number, hemoglobin content, and catalase activity were found in the blood along with higher concentrations of coproporphyrin and 17-ketosteroids. All values returned to normal after an 8-week stay of these children away from the metallurgical plant.

  1. Adsorption/oxidation of sulfur-containing gases on nitrogen-doped activated carbon

    Directory of Open Access Journals (Sweden)

    Liu Qiang

    2016-01-01

    Full Text Available Coconut shell-based activated carbon (CAC was used for the removal of methyl mercaptan (MM. CAC was modified by urea impregnation and calcined at 450°C and 950°C. The desulfurization activity was determined in a fixed bed reactor under room temperature. The results showed that the methyl mercaptan adsorption/oxidation capacity of modified carbon caicined at 950°C is more than 3 times the capacity of original samples. On the other hand, the modified carbon caicined at 950°C also has a high capacity for the simultaneous adsorption/oxidation of methyl mercaptan and hydrogen sulfide.The introduce of basic nitrogen groups siginificantly increases the desulfurization since it can facilitate the electron transfer process between sulfur and oxygen. The structure and chemical properties are characterized using Boehm titration, N2 adsorption-desorption method, thermal analysis and elemental analysis. The results showed that the major oxidation products were dimethyl disulfide and methanesulfonic acid which adsorbed in the activated carbon.

  2. From carbon nanotubes to carbon atomic chains

    Science.gov (United States)

    Casillas García, Gilberto; Zhang, Weijia; José-Yacamán, Miguel

    2010-10-01

    Carbyne is a linear allotrope of carbon. It is formed by a linear arrangement of carbon atoms with sp-hybridization. We present a reliable and reproducible experiment to obtain these carbon atomic chains using few-layer-graphene (FLG) sheets and a HRTEM. First the FLG sheets were synthesized from worm-like exfoliated graphite and then drop-casted on a lacey-carbon copper grid. Once in the TEM, two holes are opened near each other in a FLG sheet by focusing the electron beam into a small spot. Due to the radiation, the carbon atoms rearrange themselves between the two holes and form carbon fibers. The beam is concentrated on the carbon fibers in order excite the atoms and induce a tension until multi wall carbon nanotube (MWCNT) is formed. As the radiation continues the MWCNT breaks down until there is only a single wall carbon nanotube (SWCNT). Then, when the SWCNT breaks, an atomic carbon chain is formed, lasts for several seconds under the radiation and finally breaks. This demonstrates the stability of this carbon structure.

  3. Carbon nanotube composite materials

    Energy Technology Data Exchange (ETDEWEB)

    O' Bryan, Gregory; Skinner, Jack L; Vance, Andrew; Yang, Elaine Lai; Zifer, Thomas

    2015-03-24

    A material consisting essentially of a vinyl thermoplastic polymer, un-functionalized carbon nanotubes and hydroxylated carbon nanotubes dissolved in a solvent. Un-functionalized carbon nanotube concentrations up to 30 wt % and hydroxylated carbon nanotube concentrations up to 40 wt % can be used with even small concentrations of each (less than 2 wt %) useful in producing enhanced conductivity properties of formed thin films.

  4. Mutagenicity of carbon nanomaterials

    DEFF Research Database (Denmark)

    Wallin, Håkan; Jacobsen, Nicklas Raun; White, Paul A;

    2011-01-01

    Carbon nanomaterials such carbon nanotubes, graphene and fullerenes are some the most promising nanomaterials. Although carbon nanomaterials have been reported to possess genotoxic potential, it is imperitive to analyse the data on the genotoxicity of carbon nanomaterials in vivo and in vitro...

  5. Electroanalysis with carbon paste electrodes

    CERN Document Server

    Svancara, Ivan; Walcarius, Alain; Vytras, Karel

    2011-01-01

    Introduction to Electrochemistry and Electroanalysis with Carbon Paste-Based ElectrodesHistorical Survey and GlossaryField in Publication Activities and LiteratureCarbon Pastes and Carbon Paste ElectrodesCarbon Paste as the Binary MixtureClassification of Carbon Pastes and Carbon Paste ElectrodesConstruction of Carbon Paste HoldersCarbon Paste as the Electrode MaterialPhysicochemical Properties of Carbon PastesElectrochemical Characteristics of Carbon PastesTesting of Unmodified CPEsIntera

  6. Mesoporous carbon materials

    Science.gov (United States)

    Dai, Sheng; Fulvio, Pasquale Fernando; Mayes, Richard T.; Wang, Xiqing; Sun, Xiao-Guang; Guo, Bingkun

    2014-09-09

    A conductive mesoporous carbon composite comprising conductive carbon nanoparticles contained within a mesoporous carbon matrix, wherein the conductive mesoporous carbon composite possesses at least a portion of mesopores having a pore size of at least 10 nm and up to 50 nm, and wherein the mesopores are either within the mesoporous carbon matrix, or are spacings delineated by surfaces of said conductive carbon nanoparticles when said conductive carbon nanoparticles are fused with each other, or both. Methods for producing the above-described composite, devices incorporating them (e.g., lithium batteries), and methods of using them, are also described.

  7. Pyrolyzed thin film carbon

    Science.gov (United States)

    Tai, Yu-Chong (Inventor); Liger, Matthieu (Inventor); Harder, Theodore (Inventor); Konishi, Satoshi (Inventor); Miserendino, Scott (Inventor)

    2010-01-01

    A method of making carbon thin films comprises depositing a catalyst on a substrate, depositing a hydrocarbon in contact with the catalyst and pyrolyzing the hydrocarbon. A method of controlling a carbon thin film density comprises etching a cavity into a substrate, depositing a hydrocarbon into the cavity, and pyrolyzing the hydrocarbon while in the cavity to form a carbon thin film. Controlling a carbon thin film density is achieved by changing the volume of the cavity. Methods of making carbon containing patterned structures are also provided. Carbon thin films and carbon containing patterned structures can be used in NEMS, MEMS, liquid chromatography, and sensor devices.

  8. Phosphorus/sulfur Co-doped porous carbon with enhanced specific capacitance for supercapacitor and improved catalytic activity for oxygen reduction reaction

    Science.gov (United States)

    Zhou, Yao; Ma, Ruguang; Candelaria, Stephanie L.; Wang, Jiacheng; Liu, Qian; Uchaker, Evan; Li, Pengxi; Chen, Yongfang; Cao, Guozhong

    2016-05-01

    Phosphorus (P)/sulfur (S) co-doped porous carbon derived from resorcinol and furaldehyde are synthesized through one-step sol-gel processing with the addition of phosphorus pentasulfide as P and S source followed with freeze-drying and pyrolysis in nitrogen. The P/S co-doping strategy facilitates the pore size widening both in micropore and mesopore regions, together with the positive effect on the degree of graphitization of porous carbon through elimination of amorphous carbon through the formation and evaporation of carbon disulfide. As an electrode for supercapacitor application, P/S co-doped porous carbon demonstrates 43.5% improvement on specific capacitance of the single electrode compared to pristine porous carbon in organic electrolyte at a current of 0.5 mA due to the P-induced pseudocapacitive reactions. As for electrocatalytic use, promoted electrocatalytic activity and high resistance to crossover effects of oxygen reduction reaction (ORR) in alkaline media are observed after the introduction of P and S into porous carbon. After air activation, the specific capacitance of the single electrode of sample PS-pC reaches up to 103.5 F g-1 and an improved oxygen reduction current density.

  9. Improved Na Storage Performance with the Involvement of Nitrogen-Doped Conductive Carbon into WS2 Nanosheets.

    Science.gov (United States)

    Wang, Xin; Huang, Jianfeng; Li, Jiayin; Cao, Liyun; Hao, Wei; Xu, Zhanwei

    2016-09-14

    Tungsten disulfide (WS2) material is regarded as one of the most promising anode candidates for sodium ion batteries (SIBs). However, the exploration of this material still remains a great challenge to improve its cycling capacity. In this paper, nitrogen-doped conductive carbon/WS2 nanocomposites (WS2-NC) were fabricated based on the synthesis of the pure WS2 and conductive carbon/WS2 (WS2-C) nanocomposites. The reversible capacity of the as-prepared WS2-NC is stabilized at ∼360 mA h g(-1) at the density of 100 mA g(-1), even ∼200 mA h g(-1) at 1 A g(-1), presenting much better cycling performance than pure WS2 and conductive carbon/WS2 (WS2-C) samples. This excellent performance is further attributed to obviously promoted interfacial reaction in WS2 nanosheets at a low voltage platform (0.3-0.0 V), which is considered to closely relate to the incorporation of nitrogen-doped conductive carbon into WS2 nanosheets. Generally, this work presents an obviously enhanced Na storage performance by the incorporation of N-doped carbon into WS2 nanosheets to promote their interfacial reaction at low voltage platform. It could provide guidelines to create other high-capacity anode sulfide materials for SIBs. PMID:27564678

  10. Adsorption of Carbon Dioxide on Activated Carbon

    Institute of Scientific and Technical Information of China (English)

    Bo Guo; Liping Chang; Kechang Xie

    2006-01-01

    The adsorption of CO2 on a raw activated carbon A and three modified activated carbon samples B, C, and D at temperatures ranging from 303 to 333 K and the thermodynamics of adsorption have been investigated using a vacuum adsorption apparatus in order to obtain more information about the effect of CO2 on removal of organic sulfur-containing compounds in industrial gases. The active ingredients impregnated in the carbon samples show significant influence on the adsorption for CO2 and its volumes adsorbed on modified carbon samples B, C, and D are all larger than that on the raw carbon sample A. On the other hand, the physical parameters such as surface area, pore volume, and micropore volume of carbon samples show no influence on the adsorbed amount of CO2. The Dubinin-Radushkevich (D-R) equation was the best model for fitting the adsorption data on carbon samples A and B, while the Freundlich equation was the best fit for the adsorption on carbon samples C and D. The isosteric heats of adsorption on carbon samples A, B, C, and D derived from the adsorption isotherms using the Clapeyron equation decreased slightly increasing surface loading. The heat of adsorption lay between 10.5 and 28.4 kJ/mol, with the carbon sample D having the highest value at all surface coverages that were studied. The observed entropy change associated with the adsorption for the carbon samples A, B, and C (above the surface coverage of 7 ml/g) was lower than the theoretical value for mobile adsorption. However, it was higher than the theoretical value for mobile adsorption but lower than the theoretical value for localized adsorption for carbon sample D.

  11. Accelerating Mineral Carbonation Using Carbonic Anhydrase.

    Science.gov (United States)

    Power, Ian M; Harrison, Anna L; Dipple, Gregory M

    2016-03-01

    Carbonic anhydrase (CA) enzymes have gained considerable attention for their potential use in carbon dioxide (CO2) capture technologies because they are able to catalyze rapidly the interconversion of aqueous CO2 and bicarbonate. However, there are challenges for widespread implementation including the need to develop mineralization process routes for permanent carbon storage. Mineral carbonation of highly reactive feedstocks may be limited by the supply rate of CO2. This rate limitation can be directly addressed by incorporating enzyme-catalyzed CO2 hydration. This study examined the effects of bovine carbonic anhydrase (BCA) and CO2-rich gas streams on the carbonation rate of brucite [Mg(OH)2], a highly reactive mineral. Alkaline brucite slurries were amended with BCA and supplied with 10% CO2 gas while aqueous chemistry and solids were monitored throughout the experiments (hours to days). In comparison to controls, brucite carbonation using BCA was accelerated by up to 240%. Nesquehonite [MgCO3·3H2O] precipitation limited the accumulation of hydrated CO2 species, apparently preventing BCA from catalyzing the dehydration reaction. Geochemical models reproduce observed reaction progress in all experiments, revealing a linear correlation between CO2 uptake and carbonation rate. Data demonstrates that carbonation in BCA-amended reactors remained limited by CO2 supply, implying further acceleration is possible. PMID:26829491

  12. Carbon fuel cells with carbon corrosion suppression

    Science.gov (United States)

    Cooper, John F.

    2012-04-10

    An electrochemical cell apparatus that can operate as either a fuel cell or a battery includes a cathode compartment, an anode compartment operatively connected to the cathode compartment, and a carbon fuel cell section connected to the anode compartment and the cathode compartment. An effusion plate is operatively positioned adjacent the anode compartment or the cathode compartment. The effusion plate allows passage of carbon dioxide. Carbon dioxide exhaust channels are operatively positioned in the electrochemical cell to direct the carbon dioxide from the electrochemical cell.

  13. SOD1 aggregation in astrocytes following ischemia/reperfusion injury: a role of NO-mediated S-nitrosylation of protein disulfide isomerase (PDI

    Directory of Open Access Journals (Sweden)

    Chen Xueping

    2012-10-01

    Full Text Available Abstract Background Ubiquitinated-protein aggregates are implicated in cerebral ischemia/reperfusion injury. The very presence of these ubiquitinated-protein aggregates is abnormal and seems to be disease-related. However, it is not clear what leads to aggregate formation and whether the aggregations represent a reaction to aggregate-mediated neurodegeneration. Methods To study the nitrosative stress-induced protein aggregation in cerebral ischemia/reperfusion injury, we used primary astrocyte cultures as a cell model, and systematically examined their iNOS expression and consequent NO generation following oxygen glucose deprivation and reperfusion. The expression of protein disulfide isomerase (PDI and copper-zinc superoxide dismutase (SOD1 were also examined, and the biochemical interaction between PDI and SOD1 was determined by immunoprecipitation. In addition, the levels of S-nitrosylated PDI in cultured astrocytes after oxygen glucose deprivation and reperfusion treatment were measured using the biotin-switch assay. The formation of ubiquitinated-protein aggregates was detected by immunoblot and immunofluorescence staining. Results Our data showed that the up-regulation of iNOS expression after oxygen glucose deprivation and reperfusion treatment led to excessive NO generation. Up-regulation of PDI and SOD1 was also identified in cultured astrocytes following oxygen glucose deprivation and reperfusion, and these two proteins were found to bind to each other. Furthermore, the increased nitrosative stress due to ischemia/reperfusion injury was highly associated with NO-induced S-nitrosylation of PDI, and this S-nitrosylation of PDI was correlated with the formation of ubiquitinated-protein aggregates; the levels of S-nitrosylated PDI increased in parallel with the formation of aggregates. When NO generation was pharmacologically inhibited by iNOS specific inhibitor 1400W, S-nitrosylation of PDI was significantly blocked. In addition, the

  14. Dietary inclusion of diallyl disulfide, yucca powder, calcium fumarate, an extruded linseed product, or medium-chain fatty acids does not affect methane production in lactating dairy cows.

    Science.gov (United States)

    van Zijderveld, S M; Dijkstra, J; Perdok, H B; Newbold, J R; Gerrits, W J J

    2011-06-01

    Two similar experiments were conducted to assess the effect of diallyl disulfide (DADS), yucca powder (YP), calcium fumarate (CAFU), an extruded linseed product (UNSAT), or a mixture of capric and caprylic acid (MCFA) on methane production, energy balance, and dairy cow performance. In experiment 1, a control diet (CON1) and diets supplemented with 56 mg of DADS/kg of dry matter (DM), 3g of YP/kg of DM, or 25 g of CAFU/kg of DM were evaluated. In experiment 2, an inert saturated fat source in the control diet (CON2) was exchanged isolipidically for an extruded linseed source (100g/kg of DM; UNSAT) or a mixture of C8:0 and C10:0 (MCFA; 20.3g/kg of DM). In experiment 2, a higher inclusion level of DADS (200mg/kg of DM) was also tested. Both experiments were conducted using 40 lactating Holstein-Friesian dairy cows. Cows were adapted to the diet for 12 d and were subsequently kept in respiration chambers for 5 d to evaluate methane production, diet digestibility, energy balance, and animal performance. Feed intake was restricted to avoid confounding effects of possible differences in ad libitum feed intake on methane production. Feed intake was, on average, 17.5 and 16.6 kg of DM/d in experiments 1 and 2, respectively. None of the additives reduced methane production in vivo. Methane production in experiment 1 was 450, 453, 446, and 423 g/d for CON1 and the diets supplemented with DADS, YP, and CAFU, respectively. In experiment 2, methane production was 371, 394, 388, and 386 g/d for CON2 and the diets supplemented with UNSAT, MCFA, and DADS, respectively. No effects of the additives on energy balance or neutral detergent fiber digestibility were observed. The addition of MCFA increased milk fat content (5.38% vs. 4.82% for control) and fat digestibility (78.5% vs. 59.8% for control), but did not affect milk yield or other milk components. The other products did not affect milk yield or composition. Results from these experiments emphasize the need to confirm methane

  15. Folic acid-targeted disulfide-based cross-linking micelle for enhanced drug encapsulation stability and site-specific drug delivery against tumors

    Directory of Open Access Journals (Sweden)

    Zhang Y

    2016-03-01

    Full Text Available Yumin Zhang,1,* Junhui Zhou,2,* Cuihong Yang,1 Weiwei Wang,3 Liping Chu,1 Fan Huang,1 Qiang Liu,1 Liandong Deng,2 Deling Kong,3 Jianfeng Liu,1 Jinjian Liu1 1Tianjin Key Laboratory of Radiation Medicine and Molecular Nuclear Medicine, Institute of Radiation Medicine, Chinese Academy of Medical Science and Peking Union Medical College, 2Department of Polymer Science and Technology, School of Chemical Engineering and Technology, Tianjin University, 3Tianjin Key Laboratory of Biomaterial Research, Institute of Biomedical Engineering, Chinese Academy of Medical Science and Peking Union Medical College, Tianjin, People’s Republic of China *These authors contributed equally in this work Abstract: Although the shortcomings of small molecular antitumor drugs were efficiently improved by being entrapped into nanosized vehicles, premature drug release and insufficient tumor targeting demand innovative approaches that boost the stability and tumor responsiveness of drug-loaded nanocarriers. Here, we show the use of the core cross-linking method to generate a micelle with enhanced drug encapsulation ability and sensitivity of drug release in tumor. This kind of micelle could increase curcumin (Cur delivery to HeLa cells in vitro and improve tumor accumulation in vivo. We designed and synthesized the core cross-linked micelle (CCM with polyethylene glycol and folic acid-polyethylene glycol as the hydrophilic units, pyridyldisulfide as the cross-linkable and hydrophobic unit, and disulfide bond as the cross-linker. CCM showed spherical shape with a diameter of 91.2 nm by the characterization of dynamic light scattering and transmission electron microscope. Attributed to the core cross-linking, drug-loaded CCM displayed higher Nile Red or Cur-encapsulated stability and better sensitivity to glutathione than noncross-linked micelle (NCM. Cellular uptake and in vitro antitumor studies proved the enhanced endocytosis and better cytotoxicity of CCM-Cur against

  16. Interrelationships among biological activity, disulfide bonds, secondary structure, and metal ion binding for a chemically synthesized 34-amino-acid peptide derived from alpha-fetoprotein.

    Science.gov (United States)

    MacColl, R; Eisele, L E; Stack, R F; Hauer, C; Vakharia, D D; Benno, A; Kelly, W C; Mizejewski, G J

    2001-10-01

    A 34-amino-acid peptide has been chemically synthesized based on a sequence from human alpha-fetoprotein. The purified peptide is active in anti-growth assays when freshly prepared in pH 7.4 buffer at 0.20 g/l, but this peptide slowly becomes inactive. This functional change is proven by mass spectrometry to be triggered by the formation of an intrapeptide disulfide bond between the two cysteine residues on the peptide. Interpeptide cross-linking does not occur. The active and inactive forms of the peptide have almost identical secondary structures as shown by circular dichroism (CD). Zinc ions bind to the active peptide and completely prevents formation of the inactive form. Cobalt(II) ions also bind to the peptide, and the UV-Vis absorption spectrum of the cobalt-peptide complex shows that: (1) a near-UV sulfur-to-metal-ion charge-transfer band had a molar extinction coefficient consistent with two thiolate bonds to Co(II); (2) the lowest-energy visible d-d transition maximum at 659 nm, also, demonstrated that the two cysteine residues are ligands for the metal ion; (3) the d-d molar extinction coefficient showed that the metal ion-ligand complex was in a distorted tetrahedral symmetry. The peptide has two cysteines, and it is speculated that the other two metal ion ligands might be the two histidines. The Zn(II)- and Co(II)-peptide complexes had similar peptide conformations as indicated by their ultraviolet CD spectra, which differed very slightly from that of the free peptide. Surprisingly, the cobalt ions acted in the reverse of the zinc ions in that, instead of stabilizing anti-growth form of the peptide, they catalyzed its loss. Metal ion control of peptide function is a saliently interesting concept. Calcium ions, in the conditions studied, apparently do not bind to the peptide. Trifluoroethanol and temperature (60 degrees C) affected the secondary structure of the peptide, and the peptide was found capable of assuming various conformations in solution

  17. Disposition of 2-mercaptobenzothiazole and 2-mercaptobenzothiazole disulfide in rats dosed intravenously, orally, and topically and in guinea pigs dosed topically

    International Nuclear Information System (INIS)

    To determine the metabolic disposition of [14C]-2-mercaptobenzothiazole (MBT) and [14C]-2-mercaptobenzothiazole disulfide (MBTS), male and female rats were dosed topically. Topical doses were 36.1 micrograms/animal for [14C]MBT and 33.6 micrograms/animal for [14C]MBTS. Although more MBT passed through the skin than MBTS and although, relative to rats, guinea pigs absorbed a greater percentage of the dose (33.4% compared to 16.1-17.5% of the MBT and 12.2% compared to 5.94-7.87% for MBTS), the disposition of radioactivity derived from the two compounds was similar. Washing of the skin removed more of the radioactivity from guinea pigs than from rats. For both sexes of rats dosed intravenously with [14C]MBT or [14C]MBTS, disposition of the compounds was similar. In 72 h, 90.9-101% of the dose appeared in the urine and 3.79-15.1% in the feces. At this time, a small portion of the administered radioactivity remained associated with erythrocytes. Oral dosing of rats for 14 d with unlabeled MBT prior to a single dose of [14C]MBT or with unlabeled MBTS prior to a single dose of [14C]MBTS (0.730 mg/kg). For both sexes, disposition of the compounds was similar. At 96 h after dosing, a small portion of the administered radioactivity remained associated with erythrocytes, most of which was bound to the membranes. For both compounds and sexes, 60.8-101% of the radioactivity administered appeared in the urine and 3.46-9.99% in the feces in 96 h. At the time, only trace amounts of radioactivity remained in tissues other than blood. Of these tissues, thyroid contained the highest concentration. In the urine, there was a detectable MBT or MBTS, but there were two metabolites, one of which was identified as a thioglucuronide derivative of MBT. The other was possibly a sulfonic acid derivative of MBT

  18. Tungsten disulfide nanosheet and exonuclease III co-assisted amplification strategy for highly sensitive fluorescence polarization detection of DNA glycosylase activity

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Jingjin; Ma, Yefei [Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources of Education Ministry, Guangxi Normal University, Guilin, 541004 (China); Kong, Rongmei [The Key Laboratory of Life-Organic Analysis, College of Chemistry and Chemical Engineering, Qufu Normal University, Qufu, Shandong 273165 (China); Zhang, Liangliang, E-mail: liangzhang319@163.com [Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources of Education Ministry, Guangxi Normal University, Guilin, 541004 (China); Yang, Wen; Zhao, Shulin [Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources of Education Ministry, Guangxi Normal University, Guilin, 541004 (China)

    2015-08-05

    Herein, we introduced a tungsten disulfide (WS{sub 2}) nanosheet and exonuclease III (Exo III) co-assisted signal amplification strategy for highly sensitive fluorescent polarization (FP) assay of DNA glycosylase activity. Two DNA glycosylases, uracil-DNA glycosylase (UDG) and human 8-oxoG DNA glycosylase 1 (hOGG1), were tested. A hairpin-structured probe (HP) which contained damaged bases in the stem was used as the substrate. The removal of damaged bases from substrate by DNA glycosylase would lower the melting temperature of HP. The HP was then opened and hybridized with a FAM dye-labeled single strand DNA (DP), generating a duplex with a recessed 3′-terminal of DP. This design facilitated the Exo III-assisted amplification by repeating the hybridization and digestion of DP, liberating numerous FAM fluorophores which could not be adsorbed on WS{sub 2} nanosheet. Thus, the final system exhibited a small FP signal. However, in the absence of DNA glycosylases, no hybridization between DP and HP was occurred, hampering the hydrolysis of DP by Exo III. The intact DP was then adsorbed on the surface of WS{sub 2} nanosheet that greatly amplified the mass of the labeled-FAM fluorophore, resulting in a large FP value. With the co-assisted amplification strategy, the sensitivity was substantially improved. In addition, this method was applied to detect UDG activity in cell extracts. The study of the inhibition of UDG was also performed. Furthermore, this method is simple in design, easy in implementation, and selective, which holds potential applications in the DNA glycosylase related mechanism research and molecular diagnostics. - Highlights: • A fluorescence polarization strategy for DNA glycosylase activity detection was developed. • The present method was based on WS{sub 2} nanosheet and exonuclease III co-assisted signal amplification. • A high sensitivity and desirable selectivity were achieved. • This method provides a promising universal platform for DNA

  19. Production of sulfur gases and carbon dioxide by synthetic weathering of crushed drill cores from the Santa Cruz porphyry copper deposit near Casa Grande, Pinal County, Arizona

    Science.gov (United States)

    Hinkle, M.E.; Ryder, J.L.; Sutley, S.J.; Botinelly, T.

    1990-01-01

    Samples of ground drill cores from the southern part of the Santa Cruz porphyry copper deposit, Casa Grande, Arizona, were oxidized in simulated weathering experiments. The samples were also separated into various mineral fractions and analyzed for contents of metals and sulfide minerals. The principal sulfide mineral present was pyrite. Gases produced in the weathering experiments were measured by gas chromatography. Carbon dioxide, oxygen, carbonyl sulfide, sulfur dioxide and carbon disulfide were found in the gases; no hydrogen sulfide, organic sulfides, or mercaptans were detected. Oxygen concentration was very important for production of the volatiles measured; in general, oxygen concentration was more important to gas production than were metallic element content, sulfide mineral content, or mineral fraction (oxide or sulfide) of the sample. The various volatile species also appeared to be interactive; some of the volatiles measured may have been formed through gas reactions. ?? 1990.

  20. 选择性氧化形成三对二硫键合成齐考诺肽%Synthesis of Ziconotide by Selective Oxidation to Form Three Disulfide Bonds

    Institute of Scientific and Technical Information of China (English)

    曹本文; 王卫国; 李战雄; 徐红岩

    2011-01-01

    Using the different oxidation reactivity of sulfhydryls with different protective group, Ziconotide was synthesized via selective oxidation reaction. Firstly, two disulfide bonds were formatted between the 1,8,16,20-position sulfhydryls protected by Trt in the Ziconotide by air oxidation. Then,the third disulfide bond was formatted between the 15,25-position sulfhydryls protected by Acm with iodine oxidation.%利用不同保护基保护的巯基的氧化反应性不同,分别以空气和碘作为氧化剂,通过选择性氧化工艺使齐考诺肽线性肽的1,8,16,20-位两对Trt保护巯基先形成两对二硫键,然后用碘氧化15,25-位Acm保护的巯基,成功地生成第三对二硫键,合成了齐考诺肽.