WorldWideScience
 
 
1

Isolation of gamma-interferon-inducible lysosomal thiol reductase (GILT) from the Yangtze finless porpoise.  

Science.gov (United States)

In this study, we isolated the cDNA of a gamma-interferon-inducible lysosomal thiol reductase (GILT), which is critical for innate immune regulation, from the Yangtze finless porpoise (FpGILT). This gene encoded a protein with 244 amino acids and a predicted molecular weight of 28kDa. The amino acid sequence of FpGILT includes an active-site CXXC motif, a GILT signature sequence, CQHGX2ECX2NX4C, and three N-linked glycosylation sites. Phylogenetic analysis showed that FpGILT and other GILT family members were derived from a common ancestor and finless porpoises are closely related to artiodactyla. Recombinant protein (FpsGILT) was then efficiently expressed and purified, and thiol reductase activity assays suggested that FpGILT catalyses disulfide bond reduction. These findings provide a basis for understanding the characteristics of immunity in the finless porpoise and other aquatic mammals. PMID:23817143

Zhou, Lidan; Yan, Weili; Yang, Lin; Chen, Hui; Cao, Qianqian; Ren, Wenhua

2013-06-28

2

Thiol:fumarate reductase (Tfr) from Methanobacterium thermoautotrophicum--identification of the catalytic sites for fumarate reduction and thiol oxidation.  

UK PubMed Central (United Kingdom)

Most methanogenic Archaea contain an unusual cytoplasmic fumarate reductase which catalyzes the reduction of fumarate with coenzyme M (CoM-S-H) and coenzyme B (CoB-S-H) as electron donors forming succinate and CoM-S-S-CoB as products. We report here on the purification and characterization of this thiol:fumarate reductase (Tfr) from Methanobacterium thermoautotrophicum (strain Marburg). The purified enzyme, which was composed of two different subunits with apparent molecular masses of 58 kDa (TfrA) and 50 kDa (TfrB), was found to catalyze the following reactions: (a) the reduction of fumarate with CoM-S-H and CoB-S-H (150 U/mg); (b) the reduction of fumarate with reduced benzyl viologen (620 U/mg); (c) the oxidation of CoM-S-H and CoB-S-H to CoM-S-S-CoB with methylene blue (95 U/mg); and (d) the reduction of CoM-S-S-CoB with reduced benzyl viologen (250 U/mg). The flavoprotein contained 12 mol non-heme iron and approximately the same amount of acid-labile sulfur/mol heterodimer. The genes encoding TfrA and TfrB were cloned and sequenced. Sequence comparisons with fumarate reductases and succinate dehydrogenases from Bacteria and Eucarya and with heterodisulfide reductases from M. thermoautotrophicum and Methanosarcina barkeri revealed that TfrA harbors FAD-binding motifs and the catalytic site for fumarate reduction and that TfrB harbors one [2Fe-2S] cluster and two [4Fe-4S] clusters and the catalytic site for CoM-S-H and CoB-S-H oxidation.

Heim S; Künkel A; Thauer RK; Hedderich R

1998-04-01

3

Thiol:fumarate reductase (Tfr) from Methanobacterium thermoautotrophicum--identification of the catalytic sites for fumarate reduction and thiol oxidation.  

Science.gov (United States)

Most methanogenic Archaea contain an unusual cytoplasmic fumarate reductase which catalyzes the reduction of fumarate with coenzyme M (CoM-S-H) and coenzyme B (CoB-S-H) as electron donors forming succinate and CoM-S-S-CoB as products. We report here on the purification and characterization of this thiol:fumarate reductase (Tfr) from Methanobacterium thermoautotrophicum (strain Marburg). The purified enzyme, which was composed of two different subunits with apparent molecular masses of 58 kDa (TfrA) and 50 kDa (TfrB), was found to catalyze the following reactions: (a) the reduction of fumarate with CoM-S-H and CoB-S-H (150 U/mg); (b) the reduction of fumarate with reduced benzyl viologen (620 U/mg); (c) the oxidation of CoM-S-H and CoB-S-H to CoM-S-S-CoB with methylene blue (95 U/mg); and (d) the reduction of CoM-S-S-CoB with reduced benzyl viologen (250 U/mg). The flavoprotein contained 12 mol non-heme iron and approximately the same amount of acid-labile sulfur/mol heterodimer. The genes encoding TfrA and TfrB were cloned and sequenced. Sequence comparisons with fumarate reductases and succinate dehydrogenases from Bacteria and Eucarya and with heterodisulfide reductases from M. thermoautotrophicum and Methanosarcina barkeri revealed that TfrA harbors FAD-binding motifs and the catalytic site for fumarate reduction and that TfrB harbors one [2Fe-2S] cluster and two [4Fe-4S] clusters and the catalytic site for CoM-S-H and CoB-S-H oxidation. PMID:9578488

Heim, S; Künkel, A; Thauer, R K; Hedderich, R

1998-04-01

4

Thiol groups are involved in NADH-ascorbate free radical reductase activity of rat liver plasma membrane.  

UK PubMed Central (United Kingdom)

Plasma membranes purified by two-phase partition from rat liver showed an NADH-ascorbate free radical reductase activity of about 14 nmoles NADH oxidized/min/mg protein. This activity was inhibited by N-ethyl maleimide, iodoacetate and iodoacetamide, reagents that covalently block thiol groups. NADH-ascorbate free radical reductase was also inhibited by reduced glutathione and the inhibitions observed with blocking reagents and reduced compounds were additive. These results support the involvement of sulphydryl groups in NADH-AFR reductase and point out the idea that a balance between reduced sulfhydryls and oxidized disulfides is required for the optimal function of this activity, considered as part of the transplasma membrane electron transport system.

Villalba JM; Canalejo A; Burón MI; Córdoba F; Navas P

1993-04-01

5

Location of the redox-active thiols of ribonucleotide reductase: sequences similarity between the Escherichia coli and Lactobacillus leichmannii enzymes  

Energy Technology Data Exchange (ETDEWEB)

The redox-active thiols of Escherichia coli ribonucleoside diphosphate reductase and of Lactobacillus leichmannii ribonucleoside triphosphate reductase have been located by a procedure involving (1) prereduction of enzyme with dithiothreitol, (2) specific oxidation of the redox-active thiols by treatment with substrate in the absence of exogenous reductant, (3) alkylation of other thiols with iodoacetamide, and (4) reduction of the disulfides with dithiothreitol and alkylation with (1-/sup 14/C)iodoacetamide. The dithiothreitol-reduce E. coli B1 subunit is able to convert 3 equiv of CDP to dCDP and is labeled with 5.4 equiv of /sup 14/C. Sequencing of tryptic peptides shows that 2.8 equiv of /sup 14/C is on cysteines-752 and -757 at the C-terminus of B1, while 1.0-1.5 equiv of /sup 14/C is on cysteines-222 and -227. It thus appears that two sets of redox-active dithiols are involved in substrate reduction. The L. leichmannii reductase is able to convert 1.1 equiv of CTP to dCTP and is labeled with 2.1 equiv of /sup 14/C. Sequencing of tryptic peptides shows that 1.4 equiv of /sup 14/C is located on the two cysteines of C-E-G-G-A-C-P-I-K. This peptide shows remarkable and unexpected similarity to the thiol-containing region of the C-terminal peptide of E. coli B1, C-E-S-G-A-C-K-I.

Lin, A.N.I.; Ashley, G.W.; Stubbe, J.

1987-11-03

6

Location of the redox-active thiols of ribonucleotide reductase: sequences similarity between the Escherichia coli and Lactobacillus leichmannii enzymes  

International Nuclear Information System (INIS)

The redox-active thiols of Escherichia coli ribonucleoside diphosphate reductase and of Lactobacillus leichmannii ribonucleoside triphosphate reductase have been located by a procedure involving (1) prereduction of enzyme with dithiothreitol, (2) specific oxidation of the redox-active thiols by treatment with substrate in the absence of exogenous reductant, (3) alkylation of other thiols with iodoacetamide, and (4) reduction of the disulfides with dithiothreitol and alkylation with [1-14C]iodoacetamide. The dithiothreitol-reduce E. coli B1 subunit is able to convert 3 equiv of CDP to dCDP and is labeled with 5.4 equiv of 14C. Sequencing of tryptic peptides shows that 2.8 equiv of 14C is on cysteines-752 and -757 at the C-terminus of B1, while 1.0-1.5 equiv of 14C is on cysteines-222 and -227. It thus appears that two sets of redox-active dithiols are involved in substrate reduction. The L. leichmannii reductase is able to convert 1.1 equiv of CTP to dCTP and is labeled with 2.1 equiv of 14C. Sequencing of tryptic peptides shows that 1.4 equiv of 14C is located on the two cysteines of C-E-G-G-A-C-P-I-K. This peptide shows remarkable and unexpected similarity to the thiol-containing region of the C-terminal peptide of E. coli B1, C-E-S-G-A-C-K-I.

1987-01-01

7

Cloning and sequencing of thiol-specific antioxidant from mammalian brain: Alkyl hydroperoxide reductase and thiol-specific antioxidant define a large family of antioxidant enzymes  

Energy Technology Data Exchange (ETDEWEB)

A cDNA corresponding to a thiol-specific antioxidant enzyme (TSA) was isolated from a rat brain cDNA library with the use of antibodies to bovine TSA. The cDNA clone encoded an open reading frame capable of encoding a 198-residue polypeptide. The rat and yeast TSA proteins show significant sequence homology to the 21-kDa component (AhpC) of Salmonella typhimurium alkyl hydroperoxide reductase, and we have found that AhpC exhibits TSA activity. AhpC and TSA define a family of >25 different proteins present in organisms from all kingdoms. The similarity among the family members extends over the entire sequence and ranges between 23% and 98% identity. A majority of the members of the AhpC/TSA family contain two conserved cysteines. At least eight of the genes encoding AhpC/TSA-like polypeptides are found in proximity to genes encoding other oxidoreductase activities, and the expression of several of the homologs has been correlated with pathogenicity. The authors suggest that the AhpC/TSA family represents a widely distributed class of antioxidant enzymes. They also report that a second family of proteins, defined by the 57-kDa component (AhpF) of alkyl hydroperoxide reductase and by thioredoxin reductase, has expanded to include six additional members.

Chae, H.Z; Storz, G.; Rhee, S.G. [National Institutes of Health, Bethesda, MD (United States); Robison, K.; Church, G. [Harvard Medical School, Boston, MA (United States); Poole, L.B. [Bowman Gray School of Medicine of Wake Forest Univ., Winstom-Salem, NC (United States)

1994-07-19

8

Enzymatic reduction of disulfide bonds in lysosomes: Characterization of a Gamma-interferon-inducible lysosomal thiol reductase (GILT)  

Science.gov (United States)

Proteins internalized into the endocytic pathway are usually degraded. Efficient proteolysis requires denaturation, induced by acidic conditions within lysosomes, and reduction of inter- and intrachain disulfide bonds. Cytosolic reduction is mediated enzymatically by thioredoxin, but the mechanism of lysosomal reduction is unknown. We describe here a lysosomal thiol reductase optimally active at low pH and capable of catalyzing disulfide bond reduction both in vivo and in vitro. The active site, determined by mutagenesis, consists of a pair of cysteine residues separated by two amino acids, similar to other enzymes of the thioredoxin family. The enzyme is a soluble glycoprotein that is synthesized as a precursor. After delivery into the endosomal/lysosomal system by the mannose 6-phosphate receptor, N- and C-terminal prosequences are removed. The enzyme is expressed constitutively in antigen-presenting cells and induced by IFN-? in other cell types, suggesting a potentially important role in antigen processing.

Arunachalam, Balasubramanian; Phan, Uyen T.; Geuze, Hans J.; Cresswell, Peter

2000-01-01

9

A turn-on NIR fluorescence and colourimetric cyanine probe for monitoring the thiol content in serum and the glutathione reductase assisted glutathione redox process.  

UK PubMed Central (United Kingdom)

We report a novel reaction-based thiol selective turn-on near-infrared (NIR) fluorescence and colourimetric dinitrobenzenesulfonyl-cyanine (DNBSCy) probe. In the presence of thiols such as glutathione (GSH), new absorption bands (476 and 581 nm) were observed, with the colour of the solution (10 mM PBS, pH = 7.4) changing from light green to blue. Interestingly, relatively non-fluorescent DNBSCy exhibited enhanced fluorescence emission around 700 nm in the NIR region. GSH reacted efficiently with the electron withdrawing sulfonyl ester moiety of DNBSCy, releasing the quinone embedded heptamethine cyanine (Cy-quinone) with extended ?-electron conjugation responsible for the turn-on NIR fluorescence. Cy-quinone also displayed a conjugated ?-electron push–pull character under physiological conditions. The DNBSCy probe was effectively employed to monitor the thiols in fetal bovine serum (FBS). The probe was capable of monitoring the oxidized glutathione (GSSG)/GSH redox process in the presence of glutathione reductase and NADPH with NIR fluorescence and colourimetric optical response. Thus, DNBSCy has the potential to measure the activity of glutathione reductase as a measure of oxidative stress.

Maity D; Govindaraju T

2013-04-01

10

Potent in vitro anti-Trypanosoma cruzi activity of pyridine-2-thiol N-oxide metal complexes having an inhibitory effect on parasite-specific fumarate reductase.  

Science.gov (United States)

In the search for new therapeutic tools against Chagas disease (American trypanosomiasis) palladium and platinum complexes of the bioactive ligand pyridine-2-thiol N-oxide were exhaustively characterized and evaluated in vitro. Both complexes showed high in vitro growth inhibition activity (IC(50) values in the nanomolar range) against Trypanosoma cruzi, the causative agent of the disease. They were 39-115 times more active than the antitrypanosomal drug Nifurtimox. The palladium complex showed an approximately threefold enhancement of the activity compared with the parent compound. In addition, owing to their low unspecific cytotoxicity on mammalian cells, the complexes showed a highly selective antiparasite activity. To get an insight into the mechanism of action of these compounds, DNA, redox metabolism (intraparasite free-radical production) and two parasite-specific enzymes absent in the host, namely, trypanothione reductase and NADH-fumarate reductase, were evaluated as potential parasite targets. Additionally, the effect of metal coordination on the free radical scavenger capacity previously reported for the free ligand was studied. All the data strongly suggest that trypanocidal action of the complexes could mainly rely on the inhibition of the parasite-specific enzyme NADH-fumarate reductase. PMID:18322709

Vieites, Marisol; Smircich, Pablo; Parajón-Costa, Beatriz; Rodríguez, Jorge; Galaz, Verónica; Olea-Azar, Claudio; Otero, Lucía; Aguirre, Gabriela; Cerecetto, Hugo; González, Mercedes; Gómez-Barrio, Alicia; Garat, Beatriz; Gambino, Dinorah

2008-03-06

11

Identification of interferon-?-inducible-lysosomal thiol reductase (GILT) gene from Mefugu (Takifugu obscures) and its immune response to LPS challenge.  

Science.gov (United States)

Interferon-?-inducible-lysosomal thiol reductase (GILT) plays a key role in the processing and presentation of MHC class II-restricted antigen (Ag) by catalyzing disulfide bond reduction. In this study, a Mefugu cDNA (ToGILT) encodes a deduced protein of 242 amino acids with a putative molecular weight of 28.6 kDa. It contains typical features of GILT proteins including the signature sequence CQHGX2ECX2NX4C, CXXC motif and other five cysteines. Genomic analysis revealed that ToGILT gene exhibited a similar exon-intron organization to human and mouse GILT. Phylogenetic analysis showed that ToGILT derived from a common ancestor with other vertebrate GILT proteins. The ToGILT mRNA was expressed in a tissue-specific manner and obviously up-regulated in spleen and kidney after LPS induction. These results suggest that ToGILT may be involved in the immune response to bacteria challenge in Takifugu obscurus. PMID:23669023

Liu, Meng; Ai, Hongxin; Xiao, Wen; Shen, Yuefen; Shen, Yang; Cui, Xianwei; Zhang, Shuangquan

2013-05-11

12

Characterization and expression of gamma-interferon-inducible lysosomal thiol reductase (GILT) gene in rainbow trout (Oncorhynchus mykiss) with implications for GILT in innate immune response.  

UK PubMed Central (United Kingdom)

The interferon-?-inducible lysosomal thiol reductase (GILT) has been demonstrated to play an important role in the processing and presentation of MHC class II-restricted antigen (Ag) by catalyzing disulfide bond reduction. In this study, a rainbow trout cDNA (designated as rGILT) was cloned and identified from Oncorhynchus mykiss. The open reading frame of rGILT consists of 759 bases encoding a protein of 253 amino acids with an estimated molecular mass of 28.23 kDa and a theoretical isoelectric point of 4.94. The rGILT exhibited a characteristic GILT signature sequence CQHGX2ECX2NX4C and CXXC motif. Phylogenetic analysis suggested that rGILT had been derived from a common ancestor with other GILT proteins. RT-PCR results showed that rGILT and rIFN-? (rainbow trout IFN-?) mRNA was expressed in a tissue-specific manner and obviously up-regulated in splenocytes and the cells from head kidney after induction with LPS. Recombinant rGILT fused with His6 tag was efficiently expressed in Escherichia coli BL21 (DE3) and purified by Ni-NTA affinity chromatography. Further study revealed that rGILT was capable of catalyzing the reduction of the interchain disulfide bonds from intact IgG. This study shows that rGILT may be involved in the immune response to bacteria challenge and maintain first line of innate immune defense at basal level in O. mykiss. It also provides the basis for investigating on the role of GILT using O. mykiss as an animal model for related studies.

Liu M; Liu H; Guan X; Ai H; Wu H; Liu P; Gu W; Zhang S

2013-09-01

13

THIOL PROTECTING GROUP  

UK PubMed Central (United Kingdom)

The present invention relates to the use of a compound comprising a moiety of formula (I) as a reagent for protecting a thiol group in a thiol compound wherein X, X' and Y are as defined herein. The protecting group methodology of the present invention allows straightforward and selective protection of thiol groups. Cleaving of the thiol group to regenerate the thiol functional group is also facile and controllable. The present invention further provides an analogous protecting group methodology directed to protection of disulfide groups.

SMITH MARK; CADDICK STEPHEN; BAKER JAMES

14

Interactions of the antitumor drug, etoposide, with reduced thiols in vitro and in vivo.  

UK PubMed Central (United Kingdom)

The interaction of activated etoposide, 4'-demethylepipodophyllotoxin-9-(4,6-O-ethylidene-beta-D-glucopyra noside) (VP-16), with thiols has been studied both in vitro and in vivo in mice. We have found that both glutathione (GSH) and cysteine rapidly reduce the VP-16 free radical, which results in the regeneration of the parent drug and the oxidation of the thiol. Using spin-trapping and electron spin resonance (ESR) techniques, we have shown that this one-electron/hydrogen donation by thiols forms thiyl radicals (RS.) which are intermediates for the formation of the oxidized thiols. The administration of VP-16 in vivo to mice decreased the total thiol levels in liver and concomitantly increased the formation of oxidized thiols. Furthermore, VP-16 stimulated glutathione reductase in liver. While administration of VP-16 also increased the total thiol pools in kidney, in contrast, no significant effects were observed on lung and heart thiol pools.

Katki AG; Kalyanaraman B; Sinha BK

1987-01-01

15

NO, thiols and disulfides.  

UK PubMed Central (United Kingdom)

The chemical nature of the messenger molecule, nitric oxide (NO), and especially its reactivity towards thiol groups and disulfides, could explain, at least partly, its intervention in so many different biological processes. NO can be regarded as the smallest molecule suitable for electron transport in biological systems. The S-nitrosation reaction and its reverse reaction represent the most convenient general way to store, to transport and finally to release NO. Nitric oxide is also particularly convenient for playing a role in interconversions of thiol groups and disulfides in chain radical or oxidation-reduction processes, and to be subsequently engaged in complex sequences of reactions accounting for different biological situations.

Girard P; Potier P

1993-03-01

16

NO, thiols and disulfides.  

Science.gov (United States)

The chemical nature of the messenger molecule, nitric oxide (NO), and especially its reactivity towards thiol groups and disulfides, could explain, at least partly, its intervention in so many different biological processes. NO can be regarded as the smallest molecule suitable for electron transport in biological systems. The S-nitrosation reaction and its reverse reaction represent the most convenient general way to store, to transport and finally to release NO. Nitric oxide is also particularly convenient for playing a role in interconversions of thiol groups and disulfides in chain radical or oxidation-reduction processes, and to be subsequently engaged in complex sequences of reactions accounting for different biological situations. PMID:8462679

Girard, P; Potier, P

1993-03-29

17

Thiol Reactive Probes and Chemosensors  

Directory of Open Access Journals (Sweden)

Full Text Available Thiols are important molecules in the environment and in biological processes. Cysteine (Cys), homocysteine (Hcy), glutathione (GSH) and hydrogen sulfide (H2S) play critical roles in a variety of physiological and pathological processes. The selective detection of thiols using reaction-based probes and sensors is very important in basic research and in disease diagnosis. This review focuses on the design of fluorescent and colorimetric probes and sensors for thiol detection. Thiol detection methods include probes and labeling agents based on nucleophilic addition and substitution, Michael addition, disulfide bond or Se-N bond cleavage, metal-sulfur interactions and more. Probes for H2S are based on nucleophilic cyclization, reduction and metal sulfide formation. Thiol probe and chemosensor design strategies and mechanism of action are discussed in this review.

Hanjing Peng; Weixuan Chen; Yunfeng Cheng; Lovemore Hakuna; Robert Strongin; Binghe Wang

2012-01-01

18

Thiol reactive probes and chemosensors.  

Science.gov (United States)

Thiols are important molecules in the environment and in biological processes. Cysteine (Cys), homocysteine (Hcy), glutathione (GSH) and hydrogen sulfide (H(2)S) play critical roles in a variety of physiological and pathological processes. The selective detection of thiols using reaction-based probes and sensors is very important in basic research and in disease diagnosis. This review focuses on the design of fluorescent and colorimetric probes and sensors for thiol detection. Thiol detection methods include probes and labeling agents based on nucleophilic addition and substitution, Michael addition, disulfide bond or Se-N bond cleavage, metal-sulfur interactions and more. Probes for H(2)S are based on nucleophilic cyclization, reduction and metal sulfide formation. Thiol probe and chemosensor design strategies and mechanism of action are discussed in this review. PMID:23202239

Peng, Hanjing; Chen, Weixuan; Cheng, Yunfeng; Hakuna, Lovemore; Strongin, Robert; Wang, Binghe

2012-11-19

19

Interaction of selenite and tellurite with thiol-dependent redox enzymes: Kinetics and mitochondrial implications.  

UK PubMed Central (United Kingdom)

The interactions of selenite and tellurite with cytosolic and mitochondrial thioredoxin reductases (TrxR1 and TrxR2) and glutathione reductases (GR) from yeast and mammalian sources were explored. Both TrxR1 and TrxR2 act as selenite and tellurite reductases. Kinetic treatment shows that selenite has a greater affinity than tellurite with both TrxR1 and TrxR2. Considering both k(cat) and K(m), selenite shows a better catalytic efficiency than tellurite with TrxR1, whereas with TrxR2, the catalytic efficiency is similar for both chalcogens. Tellurite is a good substrate for GR, whereas selenite is almost completely ineffective. Selenite or tellurite determine a large mitochondrial permeability transition associated with thiol group oxidation. However, with increasing concentrations of both chalcogens, only about 25% of total thiols are oxidized. In isolated mitochondria, selenite or tellurite per se does not stimulate H?O? production, which, however, is increased by the presence of auranofin. They also determine a large oxidation of mitochondrial pyridine nucleotides. In ovarian cancer cells both chalcogens decrease the mitochondrial membrane potential. These results indicate that selenite and tellurite, interacting with the thiol-dependent enzymes, alter the balance connecting pyridine nucleotides and thiol redox state, consequently leading to mitochondrial and cellular alterations essentially referable to a disulfide stress.

Rigobello MP; Folda A; Citta A; Scutari G; Gandin V; Fernandes AP; Rundlöf AK; Marzano C; Björnstedt M; Bindoli A

2011-06-01

20

Quantification of Thiols and Disulfides.  

UK PubMed Central (United Kingdom)

BACKGROUND: Disulfide bond formation is a key posttranslational modification, with implications for structure, function and stability of numerous proteins. While disulfide bond formation is a necessary and essential process for many proteins, it is deleterious and disruptive for others. Cells go to great lengths to regulate thiol-disulfide bond homeostasis, typically with several, apparently redundant, systems working in parallel. Dissecting the extent of oxidation and reduction of disulfides is an ongoing challenge due, in part, to the facility of thiol/disulfide exchange reactions. SCOPE OF THE REVIEW: In the present account, we briefly survey the toolbox available to the experimentalist for the chemical determination of thiols and disulfides. We have chosen to focus on the key chemical aspects of current methodology, together with identifying potential difficulties inherent in their experimental implementation. MAJOR CONCLUSIONS: While many reagents have been described for the measurement and manipulation of the redox status of thiols and disulfides, a number of these methods remain underutilized. The ability to effectively quantify changes in redox conditions in living cells presents a continuing challenge. GENERAL SIGNIFICANCE: Many unresolved questions in the metabolic interconversion of thiols and disulfides remain. For example, while pool sizes of redox pairs and their intracellular distribution are being uncovered, very little is known about the flux in thiol-disulfide exchange pathways. New tools are needed to address this important aspect of cellular metabolism. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons.

Winther JR; Thorpe C

2013-04-01

 
 
 
 
21

Polythioethers by thiol-ene click polyaddition of ?,?-alkylene thiols.  

UK PubMed Central (United Kingdom)

The straightforward synthesis of a series of poly(thioether)s by photoinduced thiol-ene click polyaddition of ?,?-alkylene thiols is reported. It is found that linear and telechelic poly(thioether)s can be directly obtained from ?,?-alkylene thiols with, for example, alkyl chain length of m = 1,2,3, and 9. The reaction proceeds without additives such as (radical) initiators or metal compounds and can simply be carried out by UV-irradiation of the bulk monomer or monomer solution. Ex situ kinetic studies reveal that the reaction proceeds by a typical a step-growth polyaddition mechanism. As the homologue series of poly(thioether)s are now synthetically accessible, new direct pathways to tailored poly(alkyl sulphoxide)s and poly(alkyl sulfone)s are now possible.

Deubel F; Bretzler V; Holzner R; Helbich T; Nuyken O; Rieger B; Jordan R

2013-06-01

22

Thiols in a Connecticut Stratified Freshwater Lake  

Science.gov (United States)

Thiols are an important class of dissolved reduced sulfur (DRS) species in aquatic environments. They are generally formed from biological processes or during diagenesis of biogenic matter. Thiols can affect the biogeochemistry of B-type metals as they form strong complexes that influence trace metal speciation, bioavailability and toxicity. While current literature focuses on the biogeochemistry of thiols in marine systems, little is known about the biogeochemistry of thiols in oxic freshwaters. We chose to study thiols in Linsley Pond a stratified freshwater lake that has been extensively studied by Hutchinson. Our goals were to identify and quantify the range of thiols present throughout this small lake. Additionally, we hoped to discern the environmental factors that influence the production and distribution of thiols in the water column, and to evaluate importance of thiols in trace metal speciation. To identify and quantify various thiols in freshwaters, we adopted a sensitive and selective analytical method, which involves precolumn fluorometric labeling coupled to high performance liquid chromatography and sensitive fluorescence detection. Using this method, our analytical detection limit is below one nanomolar. Among others, two thiol species were observed in Linsley Pond: 3-mercaptopropionic acid (3-MPA) and glutathione (GSH). 3-MPA exists in both oxic and anoxic water layers at nanomolar levels, and increases from surface to bottom. GSH is only detected in subsurface layer and co-varies with Chl a, indicating possible biological sources of GSH in these layers. There is a third, unidentified thiol species which is currently under investigation. The unidentified thiol species appears only in anoxic lake waters, and tests indicate that it is not PC2 (phytochelatin with 2 glutamic acid-cysteine units). Throughout the water column, concentrations of all three thiols are greater in whole water samples than in the dissolved phase (0.45 um).

Hu, H.; Mylon, S. E.; Benoit, G.

2003-12-01

23

Mitochondrial Thioredoxin-Glutathione Reductase from Larval Taenia crassiceps (Cysticerci).  

UK PubMed Central (United Kingdom)

Mitochondrial thioredoxin-glutathione reductase was purified from larval Taenia crassiceps (cysticerci). The preparation showed NADPH-dependent reductase activity with either thioredoxin or GSSG, and was able to perform thiol/disulfide exchange reactions. At 25 degrees C specific activities were 437 +/- 27 mU mg(-1) and 840 +/- 49 mU mg(-1) with thioredoxin and GSSG, respectively. Apparent K(m) values were 0.87 +/- 0.04 muM, 41 +/- 6 muM and 19 +/- 10 muM for thioredoxin, GSSG and NADPH, respectively. Thioredoxin from eukaryotic sources was accepted as substrate. The enzyme reduced H(2)O(2) in a NADPH-dependent manner, although with low catalytic efficiency. In the presence of thioredoxin, mitochondrial TGR showed a thioredoxin peroxidase-like activity. All disulfide reductase activities were inhibited by auranofin, suggesting mTGR is dependent on selenocysteine. The reductase activity with GSSG showed a higher dependence on temperature as compared with the DTNB reductase activity. The variation of the GSSG- and DTNB reductase activities on pH was dependent on the disulfide substrate. Like the cytosolic isoform, mTGR showed a hysteretic kinetic behavior at moderate or high GSSG concentrations, but it was less sensitive to calcium. The enzyme was able to protect glutamine synthetase from oxidative inactivation, suggesting that mTGR is competent to contend with oxidative stress.

Guevara-Flores A; Del Arenal IP; Mendoza-Hernández G; Pardo JP; Flores-Herrera O; Rendón JL

2010-01-01

24

Controlled formation of thiol and disulfide interfaces.  

Science.gov (United States)

The work reported herein describes the controlled creation of uniform thiol-functionalized siloxane-anchored self-assembled monolayers (SAMs) and their selective transformation into intramonolayer (bridging) disulfides. These disulfides provide for the efficient immobilization of (bio)molecules bearing pendant thiols or disulfides, with no need for added oxidant. The unambiguous development of this surface chemistry required analytical methods that distinguish thiol and disulfide moieties on a surface. Physical properties such as wetting and monolayer thickness do not suffice nor do routine spectroscopic techniques (e.g., XPS, IR). Therefore, a method for distinguishing and quantifying thiol and disulfide surface functionality on a monolayer array based on the reaction with 2,4-dinitrofluorobenzene (DNFB, Sanger's reagent) is reported. DNFB readily reacts with thiol-SAMs (but not with disulfides) to form stable derivatives with distinctive IR, UV, and XPS signatures. Finally, the thiol-disulfide chemistry is applied to thiol-functionalized hybrid silica nanoparticles. These high-surface-area nanoparticles provide solid supports heavily loaded with thiol groups whose chemistry is also reported herein. PMID:23199096

Artel, Vlada; Cohen, Reut; Aped, Inbal; Ronen, Maria; Gerber, Doron; Sukenik, Chaim N

2012-12-17

25

Controlled formation of thiol and disulfide interfaces.  

UK PubMed Central (United Kingdom)

The work reported herein describes the controlled creation of uniform thiol-functionalized siloxane-anchored self-assembled monolayers (SAMs) and their selective transformation into intramonolayer (bridging) disulfides. These disulfides provide for the efficient immobilization of (bio)molecules bearing pendant thiols or disulfides, with no need for added oxidant. The unambiguous development of this surface chemistry required analytical methods that distinguish thiol and disulfide moieties on a surface. Physical properties such as wetting and monolayer thickness do not suffice nor do routine spectroscopic techniques (e.g., XPS, IR). Therefore, a method for distinguishing and quantifying thiol and disulfide surface functionality on a monolayer array based on the reaction with 2,4-dinitrofluorobenzene (DNFB, Sanger's reagent) is reported. DNFB readily reacts with thiol-SAMs (but not with disulfides) to form stable derivatives with distinctive IR, UV, and XPS signatures. Finally, the thiol-disulfide chemistry is applied to thiol-functionalized hybrid silica nanoparticles. These high-surface-area nanoparticles provide solid supports heavily loaded with thiol groups whose chemistry is also reported herein.

Artel V; Cohen R; Aped I; Ronen M; Gerber D; Sukenik CN

2013-01-01

26

Protein Thiol Modifications Visualized In Vivo  

Directory of Open Access Journals (Sweden)

Full Text Available Thiol-disulfide interconversions play a crucial role in the chemistry of biological systems. They participate in the major systems that control the cellular redox potential and prevent oxidative damage. In addition, thiol-disulfide exchange reactions serve as molecular switches in a growing number of redox-regulated proteins. We developed a differential thiol-trapping technique combined with two-dimensional gel analysis, which in combination with genetic studies, allowed us to obtain a snapshot of the in vivo thiol status of cellular proteins. We determined the redox potential of protein thiols in vivo, identified and dissected the in vivo substrate proteins of the major cellular thiol-disulfide oxidoreductases, and discovered proteins that undergo thiol modifications during oxidative stress. Under normal growth conditions most cytosolic proteins had reduced cysteines, confirming existing dogmas. Among the few partly oxidized cytosolic proteins that we detected were proteins that are known to form disulfide bond intermediates transiently during their catalytic cycle (e.g., dihydrolipoyl transacetylase and lipoamide dehydrogenase). Most proteins with highly oxidized thiols were periplasmic proteins and were found to be in vivo substrates of the disulfide-bond-forming protein DsbA. We discovered a substantial number of redox-sensitive cytoplasmic proteins, whose thiol groups were significantly oxidized in strains lacking thioredoxin A. These included detoxifying enzymes as well as many metabolic enzymes with active-site cysteines that were not known to be substrates for thioredoxin. H2O2-induced oxidative stress resulted in the specific oxidation of thiols of proteins involved in detoxification of H2O2 and of enzymes of cofactor and amino acid biosynthesis pathways such as thiolperoxidase, GTP-cyclohydrolase I, and the cobalamin-independent methionine synthase MetE. Remarkably, a number of these proteins were previously or are now shown to be redox regulated.

Leichert Lars I; Jakob Ursula

2004-01-01

27

Reaction of alkylcobalamins with thiols  

Energy Technology Data Exchange (ETDEWEB)

Carbon-13 NMR spectroscopy and phosphorus-31 NMR spectroscopy have been used to study the reaction of several alkylcobalamins with 2-mercaptoethanol. At alkaline pH, when the thiol is deprotonated, the alkyl-transfer reactions involve a nucleophilic attack of the thiolate anion on the Co-methylene carbon of the cobalamins, yielding alkyl thioethers and cob(II)alamin. In these nucleophilic displacement reactions cob(I)alamin is presumably formed as an intermediate. The higher alkylcobalamins react more slowly than methylcobalamin. The lower reactivity of ethyl- and propylcobalamin is probably the basis of the inhibition of the corrinoid-dependent methyl-transfer systems by propyl iodide. The transfer of the upper nucleoside ligand of adenosylcobalamin to 2-mercaptoethanol is a very slow process; S-adenosylmercaptoethanol and cob(II)alamin are the final products of the reaction. The dealkylation of (carboxymethyl)cobalamin is a much more facile reaction. At alkaline pH S-(carboxymethyl)mercaptoethanol and cob(II)alamin are produced, while at pH values below 8 the carbon-cobalt bond is cleaved reductively to acetate and cob(II)alamin. The reductive cleavage of the carbon-cobalt bond of (carboxymethyl)cobalamin by 2-mercaptoethanol is extremely fast when the cobalamin is in the base-off form. Because the authors have been unable to detect trans coordination of 2-mercaptoethanol, they favor a mechanism that involves a hydride attack on the Co-methylene carbon of (carboxymethyl) rather than a trans attack of the thiol on the cobalt atom.

Hogenkamp, H.P.C.; Bratt, G.T.; Kotchevar, A.T.

1987-07-28

28

Reaction of alkylcobalamins with thiols  

International Nuclear Information System (INIS)

[en] Carbon-13 NMR spectroscopy and phosphorus-31 NMR spectroscopy have been used to study the reaction of several alkylcobalamins with 2-mercaptoethanol. At alkaline pH, when the thiol is deprotonated, the alkyl-transfer reactions involve a nucleophilic attack of the thiolate anion on the Co-methylene carbon of the cobalamins, yielding alkyl thioethers and cob(II)alamin. In these nucleophilic displacement reactions cob(I)alamin is presumably formed as an intermediate. The higher alkylcobalamins react more slowly than methylcobalamin. The lower reactivity of ethyl- and propylcobalamin is probably the basis of the inhibition of the corrinoid-dependent methyl-transfer systems by propyl iodide. The transfer of the upper nucleoside ligand of adenosylcobalamin to 2-mercaptoethanol is a very slow process; S-adenosylmercaptoethanol and cob(II)alamin are the final products of the reaction. The dealkylation of (carboxymethyl)cobalamin is a much more facile reaction. At alkaline pH S-(carboxymethyl)mercaptoethanol and cob(II)alamin are produced, while at pH values below 8 the carbon-cobalt bond is cleaved reductively to acetate and cob(II)alamin. The reductive cleavage of the carbon-cobalt bond of (carboxymethyl)cobalamin by 2-mercaptoethanol is extremely fast when the cobalamin is in the base-off form. Because the authors have been unable to detect trans coordination of 2-mercaptoethanol, they favor a mechanism that involves a hydride attack on the Co-methylene carbon of (carboxymethyl) rather than a trans attack of the thiol on the cobalt atom

1987-07-28

29

The effect of copper and gallium compounds on ribonucleotide reductase  

Energy Technology Data Exchange (ETDEWEB)

The mode of action of copper complexes (CuL and CuKTS) and gallium compounds (gallium nitrate and citrate) in cytotoxicity was studied. The effects of these agents on the enzyme ribonucleotide reductase was investigated by monitoring the tyrosyl free radical present in the active site of the enzyme through electron spin resonance (ESR) spectroscopy. Ribonucleotide reductase, a key enzyme in cellular proliferation, consists of two subunits. M1, a dimer of molecular weight 170,000 contains the substrate and effector binding sites. M2, a dimer of molecular weight 88,000, contains non-heme iron and tyrosyl free radical essential for the activity of the enzyme. In studies using copper complexes, the cellular oxidative chemistry was examined by ESR studies on adduct formation with membranes, and oxidation of thiols. Membrane thiols were oxidized through the reduction of the ESR signal of the thiol adduct and the analysis of sulfhydryl content. Using the radiolabel [sup 59]Fe, the inhibitory action of copper thiosemicarbazones on cellular iron uptake was shown. The inhibitory action of CuL on ribonucleotide reductase was shown by the quenching of the tyrosyl free radical on the M2 subunit. The hypothesis that gallium directly interacts with the M2 subunit of the enzyme and displaces the iron from it was proven. The tyrosyl free radical signal from cell lysates was inhibited by the direct addition of gallium compounds. Gallium content in the cells was measured by a fluorimetric method, to ensure the presence of sufficient amounts of gallium to compete with the iron in the M2 subunit. The enzyme activity, measured by the conversion of [sup 14]C-CDP to the labeled deoxy CDP, was inhibited by the addition of gallium nitrate in a cell free assay system. The immunoprecipitation studies of the [sup 59]Fe labeled M2 protein using the monoclonal antibody directed against this subunit suggested that gallium releases iron from the M2 subunit.

Narasimhan, J.

1992-01-01

30

Protein-thiol substitution or protein dethiolation by thiol/disulfide exchange reactions: the albumin model.  

UK PubMed Central (United Kingdom)

Dethiolation experiments of thiolated albumin with thionitrobenzoic acid and thiols (glutathione, cysteine, homocysteine) were carried out to understand the role of albumin in plasma distribution of thiols and disulfide species by thiol/disulfide (SH/SS) exchange reactions. During these experiments we observed that thiolated albumin underwent thiol substitution (Alb-SS-X+RSH<-->Alb-SS-R+XSH) or dethiolation (Alb-SS-X+XSH<-->Alb-SH+XSSX), depending on the different pK(a) values of thiols involved in protein-thiol mixed disulfides (Alb-SS-X). It appeared in these reactions that the compound with lower pK(a) in mixed disulfide was a good leaving group and that the pK(a) differences dictated the kind of reaction (substitution or dethiolation). Thionitrobenzoic acid, bound to albumin by mixed disulfide (Alb-TNB), underwent rapid substitution after thiol addition, forming the corresponding Alb-SS-X (peaks at 0.25-1 min). In turn, Alb-SS-X were dethiolated by the excess nonprotein SH groups because of the lower pK(a) value in mixed disulfide with respect to that of other thiols. Dethiolation of Alb-SS-X was accompanied by formation of XSSX and Alb-SH up to equilibrium levels at 35 min, which were different for each thiol. Structures by molecular simulation of thiolated albumin, carried out for understanding the role of sulfur exposure in mixed disulfides in dethiolation process, evidenced that the sulfur exposure is important for the rate but not for determining the kind of reaction (substitution or dethiolation). Our data underline the contribution of SH/SS exchanges to determine levels of various thiols as reduced and oxidized species in human plasma.

Summa D; Spiga O; Bernini A; Venditti V; Priora R; Frosali S; Margaritis A; Di Giuseppe D; Niccolai N; Di Simplicio P

2007-11-01

31

Protein-thiol substitution or protein dethiolation by thiol/disulfide exchange reactions: the albumin model.  

Science.gov (United States)

Dethiolation experiments of thiolated albumin with thionitrobenzoic acid and thiols (glutathione, cysteine, homocysteine) were carried out to understand the role of albumin in plasma distribution of thiols and disulfide species by thiol/disulfide (SH/SS) exchange reactions. During these experiments we observed that thiolated albumin underwent thiol substitution (Alb-SS-X+RSHAlb-SS-R+XSH) or dethiolation (Alb-SS-X+XSHAlb-SH+XSSX), depending on the different pK(a) values of thiols involved in protein-thiol mixed disulfides (Alb-SS-X). It appeared in these reactions that the compound with lower pK(a) in mixed disulfide was a good leaving group and that the pK(a) differences dictated the kind of reaction (substitution or dethiolation). Thionitrobenzoic acid, bound to albumin by mixed disulfide (Alb-TNB), underwent rapid substitution after thiol addition, forming the corresponding Alb-SS-X (peaks at 0.25-1 min). In turn, Alb-SS-X were dethiolated by the excess nonprotein SH groups because of the lower pK(a) value in mixed disulfide with respect to that of other thiols. Dethiolation of Alb-SS-X was accompanied by formation of XSSX and Alb-SH up to equilibrium levels at 35 min, which were different for each thiol. Structures by molecular simulation of thiolated albumin, carried out for understanding the role of sulfur exposure in mixed disulfides in dethiolation process, evidenced that the sulfur exposure is important for the rate but not for determining the kind of reaction (substitution or dethiolation). Our data underline the contribution of SH/SS exchanges to determine levels of various thiols as reduced and oxidized species in human plasma. PMID:17607746

Summa, Domenico; Spiga, Ottavia; Bernini, Andrea; Venditti, Vincenzo; Priora, Raffaella; Frosali, Simona; Margaritis, Antonios; Di Giuseppe, Danila; Niccolai, Neri; Di Simplicio, Paolo

2007-11-01

32

Changes in plasma thiol levels induced by different phases of treatment in breast cancer; the role of commercial extract from black chokeberry.  

Science.gov (United States)

Different low-molecular-weight thiols, including glutathione, cysteine, and cysteinylglycine are physiological free radical scavengers. On the other hand, homocysteine may play a role as an oxidant. The aim of our present study was to establish in vitro the effects of the commercial extract of Aronia melanocarpa (Aronox(®)) on the amount of selected low-molecular-weight thiols and the activity of antioxidative enzymes (superoxide dismutase, glutathione peroxidase, and glutathione reductase) in plasma obtained from patients with invasive breast cancer during different phases of treatment [before or after the surgery and patients after different phases of chemotherapy (doxorubicin and cyclophosphamide)] and from healthy subjects. Patients were hospitalized in Department of Oncological Surgery and Department of Chemotherapy, Medical University of Lodz, Poland. The level of low-molecular-weight thiols was determined by high-performance liquid chromatography. We observed that in the presence of the Aronia extract changes in amount of thiols in plasma from breast cancer patients (at all tested groups) were significantly reduced. Our results showed that tested commercial extract reduced modifications of antioxidative enzymes activity in plasma from patients during different phases of treatment, but this effect was not statistical significant. Our results suggest that the Aronia extract supplementation in breast cancer patients has a beneficial effect on thiols concentration in plasma. Plasma, as reported in this work, could be used as an experimental model to evaluate the beneficial action of plant supplements, including phenolic extracts on thiols or other molecules during different phases of treatment. PMID:22949034

K?dzierska, Magdalena; G?owacki, Rafa?; Czernek, Urszula; Szyd?owska-Pazera, Katarzyna; Potemski, Piotr; Piekarski, Janusz; Jeziorski, Arkadiusz; Olas, Beata

2012-09-05

33

Changes in plasma thiol levels induced by different phases of treatment in breast cancer; the role of commercial extract from black chokeberry.  

UK PubMed Central (United Kingdom)

Different low-molecular-weight thiols, including glutathione, cysteine, and cysteinylglycine are physiological free radical scavengers. On the other hand, homocysteine may play a role as an oxidant. The aim of our present study was to establish in vitro the effects of the commercial extract of Aronia melanocarpa (Aronox(®)) on the amount of selected low-molecular-weight thiols and the activity of antioxidative enzymes (superoxide dismutase, glutathione peroxidase, and glutathione reductase) in plasma obtained from patients with invasive breast cancer during different phases of treatment [before or after the surgery and patients after different phases of chemotherapy (doxorubicin and cyclophosphamide)] and from healthy subjects. Patients were hospitalized in Department of Oncological Surgery and Department of Chemotherapy, Medical University of Lodz, Poland. The level of low-molecular-weight thiols was determined by high-performance liquid chromatography. We observed that in the presence of the Aronia extract changes in amount of thiols in plasma from breast cancer patients (at all tested groups) were significantly reduced. Our results showed that tested commercial extract reduced modifications of antioxidative enzymes activity in plasma from patients during different phases of treatment, but this effect was not statistical significant. Our results suggest that the Aronia extract supplementation in breast cancer patients has a beneficial effect on thiols concentration in plasma. Plasma, as reported in this work, could be used as an experimental model to evaluate the beneficial action of plant supplements, including phenolic extracts on thiols or other molecules during different phases of treatment.

K?dzierska M; G?owacki R; Czernek U; Szyd?owska-Pazera K; Potemski P; Piekarski J; Jeziorski A; Olas B

2013-01-01

34

Quinone Reductase 2 Is a Catechol Quinone Reductase  

Energy Technology Data Exchange (ETDEWEB)

The functions of quinone reductase 2 have eluded researchers for decades even though a genetic polymorphism is associated with various neurological disorders. Employing enzymatic studies using adrenochrome as a substrate, we show that quinone reductase 2 is specific for the reduction of adrenochrome, whereas quinone reductase 1 shows no activity. We also solved the crystal structure of quinone reductase 2 in complexes with dopamine and adrenochrome, two compounds that are structurally related to catecholamine quinones. Detailed structural analyses delineate the mechanism of quinone reductase 2 specificity toward catechol quinones in comparison with quinone reductase 1; a side-chain rotational difference between quinone reductase 1 and quinone reductase 2 of a single residue, phenylalanine 106, determines the specificity of enzymatic activities. These results infer functional differences between two homologous enzymes and indicate that quinone reductase 2 could play important roles in the regulation of catecholamine oxidation processes that may be involved in the etiology of Parkinson disease.

Fu, Yue; Buryanovskyy, Leonid; Zhang, Zhongtao (NYMEDCO)

2008-09-05

35

Genomics and X-ray microanalysis indicate that Ca2+ and thiols mediate the aggregation and adhesion of Xylella fastidiosa  

Directory of Open Access Journals (Sweden)

Full Text Available The availability of the genome sequence of the bacterial plant pathogen Xylella fastidiosa, the causal agent of citrus variegated chlorosis, is accelerating important investigations concerning its pathogenicity. Plant vessel occlusion is critical for symptom development. The objective of the present study was to search for information that would help to explain the adhesion of X. fastidiosa cells to the xylem. Scanning electron microscopy revealed that adhesion may occur without the fastidium gum, an exopolysaccharide produced by X. fastidiosa, and X-ray microanalysis demonstrated the presence of elemental sulfur both in cells grown in vitro and in cells found inside plant vessels, indicating that the sulfur signal is generated by the pathogen surface. Calcium and magnesium peaks were detected in association with sulfur in occluded vessels. We propose an explanation for the adhesion and aggregation process. Thiol groups, maintained by the enzyme peptide methionine sulfoxide reductase, could be active on the surface of the bacteria and appear to promote cell-cell aggregation by forming disulfide bonds with thiol groups on the surface of adjacent cells. The enzyme methionine sulfoxide reductase has been shown to be an auxiliary component in the adhesiveness of some human pathogens. The negative charge conferred by the ionized thiol group could of itself constitute a mechanism of adhesion by allowing the formation of divalent cation bridges between the negatively charged bacteria and predominantly negatively charged xylem walls.

B. Leite; M.L. Ishida; E. Alves; H. Carrer; S.F. Pascholati; E.W. Kitajima

2002-01-01

36

Genomics and X-ray microanalysis indicate that Ca2+ and thiols mediate the aggregation and adhesion of Xylella fastidiosa  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english The availability of the genome sequence of the bacterial plant pathogen Xylella fastidiosa, the causal agent of citrus variegated chlorosis, is accelerating important investigations concerning its pathogenicity. Plant vessel occlusion is critical for symptom development. The objective of the present study was to search for information that would help to explain the adhesion of X. fastidiosa cells to the xylem. Scanning electron microscopy revealed that adhesion may occur (more) without the fastidium gum, an exopolysaccharide produced by X. fastidiosa, and X-ray microanalysis demonstrated the presence of elemental sulfur both in cells grown in vitro and in cells found inside plant vessels, indicating that the sulfur signal is generated by the pathogen surface. Calcium and magnesium peaks were detected in association with sulfur in occluded vessels. We propose an explanation for the adhesion and aggregation process. Thiol groups, maintained by the enzyme peptide methionine sulfoxide reductase, could be active on the surface of the bacteria and appear to promote cell-cell aggregation by forming disulfide bonds with thiol groups on the surface of adjacent cells. The enzyme methionine sulfoxide reductase has been shown to be an auxiliary component in the adhesiveness of some human pathogens. The negative charge conferred by the ionized thiol group could of itself constitute a mechanism of adhesion by allowing the formation of divalent cation bridges between the negatively charged bacteria and predominantly negatively charged xylem walls.

Leite, B.; Ishida, M.L.; Alves, E.; Carrer, H.; Pascholati, S.F.; Kitajima, E.W.

2002-06-01

37

Thiol-ubiquinone assisted fragmentation of gold nanoparticles.  

UK PubMed Central (United Kingdom)

We report the spontaneous fragmentation of gold nanoparticles (AuNPs) induced, in aqueous solution at room temperature, by thiol derivative of ubiquinone, which involves the energetic electron injection from thiol-ubiquinone to the gold nanoparticles.

Riaz S; Ma W; Jing C; Nawaz MH; Li DW; Long YT

2013-02-01

38

Effect of Prolonged High-Fat Diet on Thiol-Disulfide Homeostasis in Rats  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was to determine the effect of a prolonged high-fat (HF) on thiol-disulfide homeostasis via the activity of the glutathione redox-system (GRS) in rat blood and liver. Methods: The experiment was conducted on male Wistar rats. They were divided into groups and fed on the HF diet for 30, 90 and 180 days, respectively. The HF diet consisted of beef fat and cholesterol (19 % and 2 % of the total diet, respectively). The state of the GRS was assessed in the erythrocytes and liver tissue by the glutathione, glutathione reductase (GR) and glutathione peroxidase (GP) activity. The levels of the initial and final products of lipid peroxidation – lipid hydroperoxides (LOOHs), diene conjugates (DC) and malondialdehydes (MDA) in the blood and liver were investigated. Results: Within 30 days, the HF diet inhibits the glutathione enzyme activity in the blood (GR: P<0.01; GP: P<0.001) and liver (GR, P<0.01). Within 90 days the HF diet kick-starts the beginning of the GRS compensatory response and restores the thiol-disulfide homeostasis. At 180 days, the HF diet shows failure of the compensatory processes in the glutathione system caused by the redox-imbalance in the thiol-disulfide exchange, which reveals lowered levels of glutathione, GR and GP activity (P<0.001 for all) in the blood and liver. Conclusion: Our results suggest that the thiol-disulfide status of the cells depends upon the nature of the nutrition, a long-term breach of which triggers a compensatory response and a failure of the compensatory processes in the GRS.

Yulia K. Denisenko; Tatyana P. Novgorodtseva

2013-01-01

39

An amplified assay for thiols based on reactivation of papain.  

UK PubMed Central (United Kingdom)

A sensitive spectrophotometric assay has been developed for thiol (sulfhydryl) groups using an inactive disulfide derivative of papain (papain-S-SCH3). The thiol-disulfide interchange reaction of a thiol with papain-S-SCH3 results in the stoichiometric formation of active papain (papain-SH). The reactivated papain catalyzes the hydrolysis of a chromogenic substrate, resulting in an amplified spectrophotometric signal proportional to the initial amount of thiol. A variety of thiols, e.g., cysteine, glutathione, penicillamine, cysteine methyl ester, and cysteamine, yield similar linear plots for the activity of papain vs the initial amount of thiol. An unknown concentration of a thiol is measured using a standard plot for the activity of papain vs the amount of thiol, obtained for the same thiol or for a similar thiol. Thiol groups on proteins and thiol groups of high values of pKa (2-mercaptoethanol, 3-mercaptopropanoic acid) can also be assayed using papain-S-SCH3 in the presence of excess cystamine. The assay is about 100-fold more sensitive than that using Ellman's reagent [5,5'-dithiobis(2-nitrobenzoic acid)]. A 0.4 microM solution of cysteine produces an absorbance change of 0.55 at 410 nm after 30 min in the assay, compared to a predicted change in absorbance of 0.0054 using Ellman's assay.

Singh R; Blättler WA; Collinson AR

1993-08-01

40

Oligonucleotide cyclization: the thiol-maleimide reaction revisited.  

UK PubMed Central (United Kingdom)

A novel method to synthesize cyclic oligonucleotides (5- to 26-mer) using the thiol-maleimide reaction is described. The target molecules were obtained after subsequent removal of thiol and maleimide protecting groups from 5'-maleimido-3'-thiol-derivatized linear precursors. Retro-Diels-Alder conditions deprotecting the maleimide simultaneously promoted cyclization cleanly and in high yield.

Sánchez A; Pedroso E; Grandas A

2013-01-01

 
 
 
 
41

Thiol methyltransferase activity in inflammatory bowel disease  

Digital Repository Infrastructure Vision for European Research (DRIVER)

BACKGROUND—Luminal anionic sulphide may contribute to epithelial damage in ulcerative colitis. Thiol methyltransferase (TMT) governs sulphide detoxification by the colonic mucosa and circulating erythrocytes.?AIMS—To measure levels of TMT activity in erythrocytes of surgically treated cases of colit...

Roediger, W; Babidge, W

42

THIOL AND THIOETHER STABILIZERS FOR FLUOROOLEFINS  

UK PubMed Central (United Kingdom)

The present disclosure relates to compositions comprising at least one fluoroolefin and an effective amount of a stabilizer comprising at least one thiol or thioether, or mixtures thereof. The stabilized compositions may be useful in cooling apparatus, such as refrigeration, air-conditioning, chillers and heat pumps, as well as in applications as foam blowing agents, solvents, aerosol propellants, fire extinguishants, and sterilants.

LECK THOMAS J; MINOR BARBARA HAVILAND; NAPPA MARIO JOSEPH; MOULI NANDINI C

43

Purine nucleoside phosphorylase as a cytosolic arsenate reductase.  

Science.gov (United States)

The findings of the accompanying paper (Németi and Gregus, Toxicol: Sci. 70, 4-12) indicate that the arsenate (AsV) reductase activity of rat liver cytosol is due to an SH enzyme that uses phosphate (or its analogue, arsenate, AsV) and a purine nucleoside (guanosine or inosine) as substrates. Purine nucleoside phosphorylase (PNP) is such an enzyme. It catalyzes the phosphorolytic cleavage of 6-oxopurine nucleosides according to the following scheme: guanosine (or inosine) + phosphate guanine (or hypoxanthine) + ribose-1-phosphate. Therefore, we have tested the hypothesis that PNP is responsible for the thiol- and purine nucleoside-dependent reduction of AsV to AsIII by rat liver cytosol. AsIII formed from AsV was quantified by HPLC-hydride generation-atomic fluorescence spectrometry analysis of the deproteinized incubates. The following findings support the conclusion that PNP reduces AsV to AsIII, using AsV instead of phosphate in the reaction above: (1) Specific PNP inhibitors (CI-1000, BCX-1777) at a concentration of 1 microM completely inhibited cytosolic AsV reductase activity. (2) During anion-exchange chromatography of cytosolic proteins, PNP activity perfectly coeluted with the AsV reductase activity, suggesting that both activities belong to the same protein. (3) PNP purified from calf spleen catalyzed reduction of AsV to AsIII in the presence of dithiothreitol (DTT) and a 6-oxopurine nucleoside (guanosine or inosine). (4) AsV reductase activity of purified PNP, like the cytosolic AsV reductase activity, was inhibited by phosphate (a substrate of PNP alternative to AsV), guanine and hypoxanthine (products of PNP favoring the reverse reaction), mercurial thiol reagents (nonspecific inhibitors of PNP), as well as CI-1000 and BCX-1777 (specific PNP inhibitors). Thus, PNP appears to be responsible for the AsV reductase activity of rat liver cytosol in the presence of DTT. Further research should clarify the mechanism and the in vivo significance of PNP-catalyzed reduction of AsV to AsIII. PMID:12388830

Gregus, Zoltán; Németi, Balázs

2002-11-01

44

Purine nucleoside phosphorylase as a cytosolic arsenate reductase.  

UK PubMed Central (United Kingdom)

The findings of the accompanying paper (Németi and Gregus, Toxicol: Sci. 70, 4-12) indicate that the arsenate (AsV) reductase activity of rat liver cytosol is due to an SH enzyme that uses phosphate (or its analogue, arsenate, AsV) and a purine nucleoside (guanosine or inosine) as substrates. Purine nucleoside phosphorylase (PNP) is such an enzyme. It catalyzes the phosphorolytic cleavage of 6-oxopurine nucleosides according to the following scheme: guanosine (or inosine) + phosphate <--> guanine (or hypoxanthine) + ribose-1-phosphate. Therefore, we have tested the hypothesis that PNP is responsible for the thiol- and purine nucleoside-dependent reduction of AsV to AsIII by rat liver cytosol. AsIII formed from AsV was quantified by HPLC-hydride generation-atomic fluorescence spectrometry analysis of the deproteinized incubates. The following findings support the conclusion that PNP reduces AsV to AsIII, using AsV instead of phosphate in the reaction above: (1) Specific PNP inhibitors (CI-1000, BCX-1777) at a concentration of 1 microM completely inhibited cytosolic AsV reductase activity. (2) During anion-exchange chromatography of cytosolic proteins, PNP activity perfectly coeluted with the AsV reductase activity, suggesting that both activities belong to the same protein. (3) PNP purified from calf spleen catalyzed reduction of AsV to AsIII in the presence of dithiothreitol (DTT) and a 6-oxopurine nucleoside (guanosine or inosine). (4) AsV reductase activity of purified PNP, like the cytosolic AsV reductase activity, was inhibited by phosphate (a substrate of PNP alternative to AsV), guanine and hypoxanthine (products of PNP favoring the reverse reaction), mercurial thiol reagents (nonspecific inhibitors of PNP), as well as CI-1000 and BCX-1777 (specific PNP inhibitors). Thus, PNP appears to be responsible for the AsV reductase activity of rat liver cytosol in the presence of DTT. Further research should clarify the mechanism and the in vivo significance of PNP-catalyzed reduction of AsV to AsIII.

Gregus Z; Németi B

2002-11-01

45

Evaluation of arene ruthenium(II) N-heterocyclic carbene complexes as organometallics interacting with thiol and selenol containing biomolecules.  

Science.gov (United States)

Metal complexes with N-heterocyclic carbene (NHC) ligands have been widely used in catalytic chemistry and are now increasingly considered for the development of new chemical tools and metal based drugs. Ruthenium complexes of the type (p-cymene)(NHC)RuCl(2) interacted with biologically relevant thiols and selenols, which resulted in the inhibition of enzymes such as thioredoxin reductase or cathepsin B. Pronounced antiproliferative effects could be obtained provided that an appropriate cellular uptake was achieved. Inhibition of tumor cell growth was accompanied by a perturbation of metabolic parameters such as cellular respiration. PMID:23149817

Oehninger, Luciano; Stefanopoulou, Maria; Alborzinia, Hamed; Schur, Julia; Ludewig, Stephanie; Namikawa, Kazuhiko; Muñoz-Castro, Alvaro; Köster, Reinhard W; Baumann, Knut; Wölfl, Stefan; Sheldrick, William S; Ott, Ingo

2012-11-14

46

Evaluation of arene ruthenium(II) N-heterocyclic carbene complexes as organometallics interacting with thiol and selenol containing biomolecules.  

UK PubMed Central (United Kingdom)

Metal complexes with N-heterocyclic carbene (NHC) ligands have been widely used in catalytic chemistry and are now increasingly considered for the development of new chemical tools and metal based drugs. Ruthenium complexes of the type (p-cymene)(NHC)RuCl(2) interacted with biologically relevant thiols and selenols, which resulted in the inhibition of enzymes such as thioredoxin reductase or cathepsin B. Pronounced antiproliferative effects could be obtained provided that an appropriate cellular uptake was achieved. Inhibition of tumor cell growth was accompanied by a perturbation of metabolic parameters such as cellular respiration.

Oehninger L; Stefanopoulou M; Alborzinia H; Schur J; Ludewig S; Namikawa K; Muñoz-Castro A; Köster RW; Baumann K; Wölfl S; Sheldrick WS; Ott I

2013-02-01

47

Crystal structures of two engineered thiol trypsins.  

Science.gov (United States)

We have determined the three-dimensional structures of engineered rat trypsins which mimic the active sites of two classes of cysteine proteases. The catalytic serine was replaced with cysteine (S195C) to test the ability of sulfur to function as a nucleophile in a serine protease environment. This variant mimics the cysteine trypsin class of thiol proteases. An additional mutation of the active site aspartate to an asparagine (D102N) created the catalytic triad of the papain-type cysteine proteases. Rat trypsins S195C and D102N,S195C were solved to 2.5 and 2.0 A, respectively. The refined structures were analyzed to determine the structural basis for the 10(6)-fold loss of activity of trypsin S195C and the 10(8)-fold loss of activity of trypsin D102N,S195C, relative to rat trypsin. The active site thiols were found in a reduced state in contrast to the oxidized thiols found in previous thiol protease structures. These are the first reported structures of serine proteases with the catalytic centers of sulfhydryl proteases. Structure analysis revealed only subtle global changes in enzyme conformation. The substrate binding pocket is unaltered, and active site amino acid 102 forms hydrogen bonds to H57 and S214 as well as to the backbone amides of A56 and H57. In trypsin S195C, D102 is a hydrogen-bond acceptor for H57 which allows the other imidazole nitrogen to function as a base during catalysis. In trypsin D102N,S195C, the asparagine at position 102 is a hydrogen-bond donor to H57 which places a proton on the imidazole nitrogen proximal to the nucleophile. This tautomer of H57 is unable to act as a base in catalysis.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:2611228

McGrath, M E; Wilke, M E; Higaki, J N; Craik, C S; Fletterick, R J

1989-11-28

48

Crystal structures of two engineered thiol trypsins.  

UK PubMed Central (United Kingdom)

We have determined the three-dimensional structures of engineered rat trypsins which mimic the active sites of two classes of cysteine proteases. The catalytic serine was replaced with cysteine (S195C) to test the ability of sulfur to function as a nucleophile in a serine protease environment. This variant mimics the cysteine trypsin class of thiol proteases. An additional mutation of the active site aspartate to an asparagine (D102N) created the catalytic triad of the papain-type cysteine proteases. Rat trypsins S195C and D102N,S195C were solved to 2.5 and 2.0 A, respectively. The refined structures were analyzed to determine the structural basis for the 10(6)-fold loss of activity of trypsin S195C and the 10(8)-fold loss of activity of trypsin D102N,S195C, relative to rat trypsin. The active site thiols were found in a reduced state in contrast to the oxidized thiols found in previous thiol protease structures. These are the first reported structures of serine proteases with the catalytic centers of sulfhydryl proteases. Structure analysis revealed only subtle global changes in enzyme conformation. The substrate binding pocket is unaltered, and active site amino acid 102 forms hydrogen bonds to H57 and S214 as well as to the backbone amides of A56 and H57. In trypsin S195C, D102 is a hydrogen-bond acceptor for H57 which allows the other imidazole nitrogen to function as a base during catalysis. In trypsin D102N,S195C, the asparagine at position 102 is a hydrogen-bond donor to H57 which places a proton on the imidazole nitrogen proximal to the nucleophile. This tautomer of H57 is unable to act as a base in catalysis.(ABSTRACT TRUNCATED AT 250 WORDS)

McGrath ME; Wilke ME; Higaki JN; Craik CS; Fletterick RJ

1989-11-01

49

Glutaredoxins in thiol/disulfide exchange.  

UK PubMed Central (United Kingdom)

SIGNIFICANCE: Glutaredoxins (Grxs) are small oxidoreductases of the thioredoxin family of proteins regulating the thiol redox state of several proteins. Thereby, Grxs are key elements in redox signaling. RECENT ADVANCES: Redox signaling via protein thiols depends on reversible oxidative modifications induced mainly by reactive oxygen/nitrogen species and glutathione (GSH) in form of its oxidized disulfide or S-nitroso-glutathione. Grxs contribute to redox signaling by the catalysis of glutathionylation, de-glutathionylation, as well as reduction of disulfide bridges via two distinct enzymatic mechanisms. The dithiol mechanism utilizes both active site cysteines to reduce disulfides, whereas the monothiol mechanism utilizes only the N-terminal active site cysteine for the reduction of GSH mixed disulfides. The sphere of action of Grxs continues to grow with the recent identification of novel targets. CRITICAL ISSUES: Because of limited methodological tools, the identification of new substrates for oxidoreductases in general is one of the biggest challenges in this research area. FUTURE DIRECTIONS: With this review, we provide a condensed summary of the current knowledge of thiol/disulfide exchange reactions catalyzed by Grxs regarding the mechanistic, structural, and functional aspects. The latter will be of high importance for future research directions, gaining novel insights into redox signaling in general, and the role of Grxs in particular.

Lillig CH; Berndt C

2013-05-01

50

Thiols and antioxidants in radiobiology: chemical and bioanalytical problems  

International Nuclear Information System (INIS)

Thiols may radioprotect by donating hydrogen atoms to a carbon-centred radical, but ascorbate (a better electron- than hydrogen-donor) can protect under some circumstances. The thiyl radical which may be produced is also reactive. Radioprotective efficiency may reflect both chemical reactivity and accessibility to DNA. Differences in uptake of thiols and thiol/disulphide exchange necessitate as much attention to chemical analysis of the test system as to its radiobiology. (author).

1991-01-01

51

Functional and Structural Characterization of a Thiol Peroxidase from Mycobacterium tuberculosis  

Energy Technology Data Exchange (ETDEWEB)

A thiol peroxidase (Tpx) from Mycobacterium tuberculosis was functionally analyzed. The enzyme shows NADPH-linked peroxidase activity using a thioredoxin-thioredoxin reductase system as electron donor, and anti-oxidant activity in a thiol-dependent metal-catalyzed oxidation system. It reduces H{sub 2}O{sub 2}, t-butyl hydroperoxide, and cumene hydroperoxide, and is inhibited by sulfhydryl reagents. Mutational studies revealed that the peroxidatic (Cys60) and resolving (Cys93) cysteine residues are critical amino acids for catalytic activity. The X-ray structure determined to a resolution of 1.75 Angstroms shows a thioredoxin fold similar to that of other peroxiredoxin family members. Superposition with structural homologues in oxidized and reduced forms indicates that the M. tuberculosis Tpx is a member of the atypical two-Cys peroxiredoxin family. In addition, the short distance that separates the Ca atoms of Cys60 and Cys93 and the location of these cysteine residues in unstructured regions may indicate that the M. tuberculosis enzyme is oxidized, though the side-chain of Cys60 is poorly visible. It is solely in the reduced Streptococcus pneumoniae Tpx structure that both residues are part of two distinct helical segments. The M. tuberculosis Tpx is dimeric both in solution and in the crystal structure. Amino acid residues from both monomers delineate the active site pocket.

Rho,B.; Hung, L.; Holton, J.; Vigil, D.; Kim, S.; Park, M.; Terwilliger, T.; Pedelacq, j.

2006-01-01

52

Sequential Opening of Mitochondrial Ion Channels as a Function of Glutathione Redox Thiol Status*s  

Science.gov (United States)

Mitochondrial membrane potential (??m) depolarization contributes to cell death and electrical and contractile dysfunction in the post-ischemic heart. An imbalance between mitochondrial reactive oxygen species production and scavenging was previously implicated in the activation of an inner membrane anion channel (IMAC), distinct from the permeability transition pore (PTP), as the first response to metabolic stress in cardiomyocytes. The glutathione redox couple, GSH/GSSG, oscillated in parallel with ??m and the NADH/NAD+ redox state. Here we show that depletion of reduced glutathione is an alternative trigger of synchronized mitochondrial oscillation in cardiomyocytes and that intermediate GSH/GSSG ratios cause reversible ??m depolarization, although irreversible PTP activation is induced by extensive thiol oxidation. Mitochondrial dysfunction in response to diamide occurred in stages, progressing from oscillations in ??m to sustained depolarization, in association with depletion of GSH. Mitochondrial oscillations were abrogated by 4?-chlorodiazepam, an IMAC inhibitor, whereas cyclosporin A was ineffective. In saponin-permeabilized cardiomyocytes, the thiol redox status was systematically clamped at GSH/GSSG ratios ranging from 300:1 to 20:1. At ratios of 150:1-100:1, ??m depolarized reversibly, and a matrix-localized fluorescent marker was retained; however, decreasing the GSH/GSSG to 50:1 irreversibly depolarized ??m and induced maximal rates of reactive oxygen species production, NAD(P)H oxidation, and loss of matrix constituents. Mitochondrial GSH sensitivity was altered by inhibiting either GSH uptake, the NADPH-dependent glutathione reductase, or the NADH/NADPH transhydrogenase, indicating that matrix GSH regeneration or replenishment was crucial. The results indicate that GSH/GSSG redox status governs the sequential opening of mitochondrial ion channels (IMAC before PTP) triggered by thiol oxidation in cardiomyocytes.

Aon, Miguel A.; Cortassa, Sonia; Maack, Christoph; O'Rourke, Brian

2008-01-01

53

Sequential opening of mitochondrial ion channels as a function of glutathione redox thiol status.  

Science.gov (United States)

Mitochondrial membrane potential (DeltaPsi(m)) depolarization contributes to cell death and electrical and contractile dysfunction in the post-ischemic heart. An imbalance between mitochondrial reactive oxygen species production and scavenging was previously implicated in the activation of an inner membrane anion channel (IMAC), distinct from the permeability transition pore (PTP), as the first response to metabolic stress in cardiomyocytes. The glutathione redox couple, GSH/GSSG, oscillated in parallel with DeltaPsi(m) and the NADH/NAD(+) redox state. Here we show that depletion of reduced glutathione is an alternative trigger of synchronized mitochondrial oscillation in cardiomyocytes and that intermediate GSH/GSSG ratios cause reversible DeltaPsi(m) depolarization, although irreversible PTP activation is induced by extensive thiol oxidation. Mitochondrial dysfunction in response to diamide occurred in stages, progressing from oscillations in DeltaPsi(m) to sustained depolarization, in association with depletion of GSH. Mitochondrial oscillations were abrogated by 4'-chlorodiazepam, an IMAC inhibitor, whereas cyclosporin A was ineffective. In saponin-permeabilized cardiomyocytes, the thiol redox status was systematically clamped at GSH/GSSG ratios ranging from 300:1 to 20:1. At ratios of 150:1-100:1, DeltaPsi(m) depolarized reversibly, and a matrix-localized fluorescent marker was retained; however, decreasing the GSH/GSSG to 50:1 irreversibly depolarized DeltaPsi(m) and induced maximal rates of reactive oxygen species production, NAD(P)H oxidation, and loss of matrix constituents. Mitochondrial GSH sensitivity was altered by inhibiting either GSH uptake, the NADPH-dependent glutathione reductase, or the NADH/NADPH transhydrogenase, indicating that matrix GSH regeneration or replenishment was crucial. The results indicate that GSH/GSSG redox status governs the sequential opening of mitochondrial ion channels (IMAC before PTP) triggered by thiol oxidation in cardiomyocytes. PMID:17540766

Aon, Miguel A; Cortassa, Sonia; Maack, Christoph; O'Rourke, Brian

2007-05-31

54

Nitrate Reductase from Monoraphidium braunii  

Science.gov (United States)

Homogeneous nitrate reductase (EC 1.6.6.2) from Monoraphidium braunii was obtained by means of affinity chromatography in blue-Sepharose and gel filtration. After electrophoresis in polyacrylamide, gel slices containing pure nitrate reductase were disrupted and injected into previously unimmunized rabbits. The antiserum produced after several weeks was found to inhibit the different activities of nitrate reductase to a similar degree. Monospecificity of the antiserum was demonstrated by Ouchterlony double diffusion and crossed immunoelectrophoresis. The antibodies were purified by immunoabsorption to Sepharose-bound nitrate reductase. The intracellular location of nitrate reductase in green algae was examined by applying an immunocytochemical method to M. braunii cells. Ultrathin frozen sections were first treated with immunopurified anti-nitrate reductase monospecific antibodies, followed by incubation with colloidal gold-labeled goat antirabbit immunoglobulin G as a marker. The enzyme was specifically located in the pyrenoid region of the chloroplast. Images Fig. 1 Fig. 2 Fig. 4 Fig. 5

Lopez-Ruiz, Antonio; Roldan, Jose Manuel; Verbelen, Jean Pierre; Diez, Jesus

1985-01-01

55

Characterization of two active site mutations of thioredoxin reductase from Escherichia coli.  

UK PubMed Central (United Kingdom)

Thioredoxin reductase (TRR), a member of the pyridine nucleotide-disulfide oxidoreductase family of flavoenzymes, undergoes two sequential thiol-disulfide interchange reactions with thioredoxin during catalysis. In order to assess the catalytic role of each nascent thiol of the active site disulfide of thioredoxin reductase, the 2 cysteines (Cys-136 and Cys-139) forming this disulfide have been individually changed to serines by site-directed mutageneses of the cloned trxB gene of Escherichia coli. Spectral analyses of TRR(Ser-136,Cys-139) as a function of pH and ionic strength have revealed two pKa values associated with the epsilon 456, one of which increases from 7.0 to 8.3 as the ionic strength is increased, and a second at 4.4 which is seen only at high ionic strength. epsilon 458 of wild type TRR(Cys-136,Cys-139) and epsilon 453 of TRR(Cys-136,Ser-139) are pH-independent. A charge transfer complex (epsilon 530 = 1300 M-1 cm-1), unique to TRR(Ser-136,Cys-139), has been observed under conditions of high ammonium cation concentration (apparent Kd = 54 microM) at pH 7.6. These results suggest the assignment of Cys-139 as the FAD-interacting thiol in the reduction of thioredoxin by NADPH via thioredoxin reductase. If, as with other members of this enzyme family, the two distinct catalytic functions are each carried out by a different nascent thiol, then Cys-136 would perform the initial thiol-disulfide interchange with thioredoxin. Steady state kinetic analyses of the proteins have revealed turnover numbers of 10 and 50% of the value of the wild type enzyme for TRR(Ser-136,Cys-139) and TRR(Cys-136,Ser-139), respectively, and no changes in the apparent Km values of TR(S2) or NADPH. The finding of activity in the mutants indicates that the remaining thiol can carry out interchange with the disulfide of thioredoxin, and the resulting mixed disulfide can be reduced by NADPH via the flavin.

Prongay AJ; Engelke DR; Williams CH Jr

1989-02-01

56

Role of endogenous thiols in protection  

Science.gov (United States)

Aminothiols represent the most important group of radioprotective compounds. The most effective compounds administered at an optimal dose and time before irradiation are able to provide a protection in mice with a dose reduction factor (DRF) of about 2-2.5. The working mechanism can partly be explained as a scavenging process of radicals induced in water and partly as a chemical repair process of injured DNA. The endogenous aminothiol which has far-out the highest intracellular concentration is glutathione (GSH). The importance of intracellular GSH in determining cellular radiosensitivity has been shown by irradiating cells that had very low GSH levels. Such cells appear to have a high radiosensitivity, especially in hypoxic conditions. On the other hand, it has been demonstrated that induction of a high GSH level (100-200% above the normal level) provides only a small protection. In vitro experiments with DNA indicate that thiols with a high positive charge condense in the vicinity of DNA and are effective protectors, whereas thiols with a negative charge are kep away from it and are poor protectors. In comparison with the most effective exogenous aminothiols like cysteamine and WR1065, GSH is not an effective radioprotector. Putative explanations for this relatively poor protective ability of GSH are presented.

Vos, O.

57

Transition metal photoredox catalysis of radical thiol-ene reactions.  

UK PubMed Central (United Kingdom)

We describe the anti-Markovnikov hydrothiolation of olefins using visible-light-absorbing transition metal photocatalysts. The key thiyl radical intermediates are generated upon quenching of photoexcited Ru*(bpz)3(2) with a variety of thiols. The adducts of a wide variety of olefins and thiols are formed in excellent yield (73-99%).

Tyson EL; Ament MS; Yoon TP

2013-03-01

58

Stereoselective synthesis of ?-glycosyl thiols and their synthetic applications.  

UK PubMed Central (United Kingdom)

A significantly fast reaction condition for the exclusive preparation ?-glycosyl thiol derivatives has been developed successfully. The reaction condition is one-step, fast, high yielding, highly stereoselective, and requires only benchtop chemicals. Further reaction of glycosyl thiol derivatives with Michael acceptors and alkylating agents furnished thioglycosides and (1,1)-thiolinked trehalose analogs.

Jana M; Misra AK

2013-03-01

59

Studies on alterations of the 86-rubidium efflux from rat pancreatic islets caused by thiol and thiol oxidants  

International Nuclear Information System (INIS)

[en] The following findings were revealed by this study: 1) Oxidation-reduction (redox) of the intracellular system of glutathione influences the potassium efflux by way of an increase in the 86-rubidium efflux brought about by the oxidation of intracellular thiols. 2) The 86-rubidium efflux is not subject to change by oxidation of extracellular thiols located in the membrane, nor can it in any way be influenced by reduced glutathione of exogenous origin. 3) The potassium efflux from rat pancreatic islets, being generally known to trigger the electric activities of the beta-cell, is controlled by the oxidation-reduction of intracellular thiols rather than by that of extracellular thiols. (TRV)[de] Der Autor dieser Arbeit kommt zu folgenden Ergebnissen: 1. Der Redoxzustand des intrazellulaeren Systems von Glutathion in reduzierter und oxidierter Form besitzt einen Einfluss auf den Kaliumefflux, wobei die Oxidation intrazellulaerer Thiole zu einer Steigerung des 86-Rubidiumeffluxes fuehrt. 2. Der 86-Rubidiumefflux wird nicht von der Oxidation extrazellulaerer, membranstaendiger Thiole beeinflusst, ebensowenig von der exogenen Zufuhr reduzierten Glutathions. 3. Das Kaliumefflux der Langerhans'schen Inseln der Ratte - als Ausloesemechanismus fuer die elektrischen Aktivitaeten der ?-Zelle - unterliegt der Regulation durch den Redoxzustand intrazellulaerer Thiole, jedoch nicht durch den Redoxzustand extrazellulaerer Thiole. (TRV)

1983-01-01

60

Human brain aldehyde reductases: relationship to succinic semialdehyde reductase and aldose reductase.  

Science.gov (United States)

Human brain contains multiple forms of aldehyde-reducing enzymes. One major form (AR3), as previously shown, has properties that indicate its identity with NADPH-dependent aldehyde reductase isolated from brain and other organs of various species; i.e., low molecular weight, use of NADPH as the preferred cofactor, and sensitivity to inhibition by barbiturates. A second form of aldehyde reductase ("SSA reductase") specifically reduces succinic semialdehyde (SSA) to produce gamma-hydroxybutyrate. This enzyme form has a higher molecular weight than AR3, and uses NADH as well as NADPH as cofactor. SSA reductase was not inhibited by pyrazole, oxalate, or barbiturates, and the only effective inhibitor found was the flavonoid quercetine. Although AR3 can also reduce SSA, the relative specificity of SSA reductase may enhance its in vivo role. A third form of human brain aldehyde reductase, AR2, appears to be comparable to aldose reductases characterized in several species, on the basis of its activity pattern with various sugar aldehydes and its response to characteristic inhibitors and activators, as well as kinetic parameters. This enzyme is also the most active in reducing the aldehyde derivatives of biogenic amines. These studies suggest that the various forms of human brain aldehyde reductases may have specific physiological functions. PMID:6778961

Hoffman, P L; Wermuth, B; von Wartburg, J P

1980-08-01

 
 
 
 
61

Human brain aldehyde reductases: relationship to succinic semialdehyde reductase and aldose reductase.  

UK PubMed Central (United Kingdom)

Human brain contains multiple forms of aldehyde-reducing enzymes. One major form (AR3), as previously shown, has properties that indicate its identity with NADPH-dependent aldehyde reductase isolated from brain and other organs of various species; i.e., low molecular weight, use of NADPH as the preferred cofactor, and sensitivity to inhibition by barbiturates. A second form of aldehyde reductase ("SSA reductase") specifically reduces succinic semialdehyde (SSA) to produce gamma-hydroxybutyrate. This enzyme form has a higher molecular weight than AR3, and uses NADH as well as NADPH as cofactor. SSA reductase was not inhibited by pyrazole, oxalate, or barbiturates, and the only effective inhibitor found was the flavonoid quercetine. Although AR3 can also reduce SSA, the relative specificity of SSA reductase may enhance its in vivo role. A third form of human brain aldehyde reductase, AR2, appears to be comparable to aldose reductases characterized in several species, on the basis of its activity pattern with various sugar aldehydes and its response to characteristic inhibitors and activators, as well as kinetic parameters. This enzyme is also the most active in reducing the aldehyde derivatives of biogenic amines. These studies suggest that the various forms of human brain aldehyde reductases may have specific physiological functions.

Hoffman PL; Wermuth B; von Wartburg JP

1980-08-01

62

A highly chemoselective and practical alkynylation of thiols.  

Science.gov (United States)

A thiol-alkynylation procedure utilizing the hypervalent iodine alkyne transfer reagent TIPS-ethynyl-benziodoxolone has been developed. This scalable reaction proceeds in five minutes at room temperature in an open flask using commercially available reagents. The scope of the reaction is broad, with a variety of phenolic, benzylic, heterocyclic, and aliphatic thiols undergoing alkynylation in excellent yield. The method is highly chemoselective as a vast array of functional groups are tolerated. The utility of the thiol-alkynylation in postsynthetic elaboration has been demonstrated through the facile installment of a fluorophore tag on a cysteine-containing peptide. PMID:23777551

Frei, Reto; Waser, Jérôme

2013-06-18

63

A highly chemoselective and practical alkynylation of thiols.  

UK PubMed Central (United Kingdom)

A thiol-alkynylation procedure utilizing the hypervalent iodine alkyne transfer reagent TIPS-ethynyl-benziodoxolone has been developed. This scalable reaction proceeds in five minutes at room temperature in an open flask using commercially available reagents. The scope of the reaction is broad, with a variety of phenolic, benzylic, heterocyclic, and aliphatic thiols undergoing alkynylation in excellent yield. The method is highly chemoselective as a vast array of functional groups are tolerated. The utility of the thiol-alkynylation in postsynthetic elaboration has been demonstrated through the facile installment of a fluorophore tag on a cysteine-containing peptide.

Frei R; Waser J

2013-07-01

64

Benzimidazol-2-ylidene gold(I) complexes are thioredoxin reductase inhibitors with multiple antitumor properties.  

UK PubMed Central (United Kingdom)

Gold(I) complexes such as auranofin have been used for decades to treat symptoms of rheumatoid arthritis and have also demonstrated a considerable potential as new anticancer drugs. The enzyme thioredoxin reductase (TrxR) is considered as the most relevant molecular target for these species. The here investigated gold(I) complexes with benzimidazole derived N-heterocyclic carbene (NHC) ligands represent a promising class of gold coordination compounds with a good stability against the thiol glutathione. TrxR was selectively inhibited by in comparison to the closely related enzyme glutathione reductase, and all complexes triggered significant antiproliferative effects in cultured tumor cells. More detailed studies on a selected complex revealed a distinct pharmacodynamic profile including the high increase of reactive oxygen species formation, apoptosis induction, strong effects on cellular metabolism (related to cell surface properties, respiration, and glycolysis), inhibition of mitochondrial respiration and activity against resistant cell lines.

Rubbiani R; Kitanovic I; Alborzinia H; Can S; Kitanovic A; Onambele LA; Stefanopoulou M; Geldmacher Y; Sheldrick WS; Wolber G; Prokop A; Wölfl S; Ott I

2010-12-01

65

Thioredoxin glutathione reductase-dependent redox networks in platyhelminth parasites.  

Science.gov (United States)

Abstract Significance: Platyhelminth parasites cause chronic infections that are a major cause of disability, mortality, and economic losses in developing countries. Maintaining redox homeostasis is a major adaptive problem faced by parasites and its disruption can shift the biochemical balance toward the host. Platyhelminth parasites possess a streamlined thiol-based redox system in which a single enzyme, thioredoxin glutathione reductase (TGR), a fusion of a glutaredoxin (Grx) domain to canonical thioredoxin reductase (TR) domains, supplies electrons to oxidized glutathione (GSSG) and thioredoxin (Trx). TGR has been validated as a drug target for schistosomiasis. Recent Advances: In addition to glutathione (GSH) and Trx reduction, TGR supports GSH-independent deglutathionylation conferring an additional advantage to the TGR redox array. Biochemical and structural studies have shown that the TR activity does not require the Grx domain, while the glutathione reductase and deglutathionylase activities depend on the Grx domain, which receives electrons from the TR domains. The search for TGR inhibitors has identified promising drug leads, notably oxadiazole N-oxides. Critical Issues: A conspicuous feature of platyhelminth TGRs is that their Grx-dependent activities are temporarily inhibited at high GSSG concentrations. The mechanism underlying the phenomenon and its biological relevance are not completely understood. Future Directions: The functional diversity of Trxs and Grxs encoded in platyhelminth genomes remains to be further assessed to thoroughly understand the TGR-dependent redox network. Optimization of TGR inhibitors and identification of compounds targeting other parasite redox enzymes are good options to clinically develop relevant drugs for these neglected, but important diseases. Antioxid. Redox Signal. 19, 735-745. PMID:22909029

Williams, David L; Bonilla, Mariana; Gladyshev, Vadim N; Salinas, Gustavo

2012-10-03

66

Thioredoxin glutathione reductase-dependent redox networks in platyhelminth parasites.  

UK PubMed Central (United Kingdom)

Abstract Significance: Platyhelminth parasites cause chronic infections that are a major cause of disability, mortality, and economic losses in developing countries. Maintaining redox homeostasis is a major adaptive problem faced by parasites and its disruption can shift the biochemical balance toward the host. Platyhelminth parasites possess a streamlined thiol-based redox system in which a single enzyme, thioredoxin glutathione reductase (TGR), a fusion of a glutaredoxin (Grx) domain to canonical thioredoxin reductase (TR) domains, supplies electrons to oxidized glutathione (GSSG) and thioredoxin (Trx). TGR has been validated as a drug target for schistosomiasis. Recent Advances: In addition to glutathione (GSH) and Trx reduction, TGR supports GSH-independent deglutathionylation conferring an additional advantage to the TGR redox array. Biochemical and structural studies have shown that the TR activity does not require the Grx domain, while the glutathione reductase and deglutathionylase activities depend on the Grx domain, which receives electrons from the TR domains. The search for TGR inhibitors has identified promising drug leads, notably oxadiazole N-oxides. Critical Issues: A conspicuous feature of platyhelminth TGRs is that their Grx-dependent activities are temporarily inhibited at high GSSG concentrations. The mechanism underlying the phenomenon and its biological relevance are not completely understood. Future Directions: The functional diversity of Trxs and Grxs encoded in platyhelminth genomes remains to be further assessed to thoroughly understand the TGR-dependent redox network. Optimization of TGR inhibitors and identification of compounds targeting other parasite redox enzymes are good options to clinically develop relevant drugs for these neglected, but important diseases. Antioxid. Redox Signal. 19, 735-745.

Williams DL; Bonilla M; Gladyshev VN; Salinas G

2013-09-01

67

Antioxidative Effects of Garlic and Some Thiol-Containing Compounds  

Directory of Open Access Journals (Sweden)

Full Text Available The term thiol refers to compounds containing sulfur. Thiol-containing compounds are found in all body cells and are indispensable for life. Some of these include cysteine, methionine, taurine, glutathione, lipoic acid, mercaptopropionylglycine, N-acetylcysteine, and the three major organosulfur compounds of garlic oil, namely, diallylsulfide, diallyldisulfide, and diallyltrisulfide. With respect to structure-function relationship, dihydrolipoic acid is the most effective antioxidant. Dihydrolipoic acid has two sulfhydryl groups and it can undergo further oxidation reaction forming lipoic acid. The more highly reduced form a thiol-containing compound has, the more stronger antioxidative activity it exerts. The number of sulfur atoms determines, at least in part, the modulatory activity on the glutathione-related antioxidant enzymes. In this article, antioxidant and antioxidative effects of garlic and some thiol-containing compounds are reviewed.

Gulizar ATMACA

2003-01-01

68

Interfacial thiol-ene photoclick reactions for forming multilayer hydrogels.  

UK PubMed Central (United Kingdom)

Interfacial visible light-mediated thiol-ene photoclick reactions were developed for preparing step-growth hydrogels with multilayer structures. The effect of a noncleavage type photoinitiator eosin-Y on visible-light-mediated thiol-ene photopolymerization was first characterized using in situ photorheometry, gel fraction, and equilibrium swelling ratio. Next, spectrophotometric properties of eosin-Y in the presence of various relevant macromer species were evaluated using ultraviolet-visible light (UV-vis) spectrometry. It was determined that eosin-Y was able to reinitiate the thiol-ene photoclick reaction, even after light exposure. Because of its small molecular weight, most eosin-Y molecules readily leached out from the hydrogels. The diffusion of residual eosin-Y from preformed hydrogels was exploited for fabricating multilayer step-growth hydrogels. Interfacial hydrogel coating was formed via the same visible-light-mediated gelation mechanism without adding fresh initiator. The thickness of the thiol-ene gel coating could be easily controlled by adjusting visible light exposure time, eosin-Y concentration initially loaded in the core gel, or macromer concentration in the coating solution. The major benefits of this interfacial thiol-ene coating system include its simplicity and cytocompatibility. The formation of thiol-ene hydrogels and coatings neither requires nor generates any cytotoxic components. This new gelation chemistry may have great utilities in controlled release of multiple sensitive growth factors and encapsulation of multiple cell types for tissue regeneration.

Shih H; Fraser AK; Lin CC

2013-03-01

69

Interaction Profile of Diphenyl Diselenide with Pharmacologically Significant Thiols  

Directory of Open Access Journals (Sweden)

Full Text Available Diphenyl diselenide has shown interesting biological activities in various free-radical-induced damage models and can be considered as a potential candidate drug against oxidative stress. Apart from its anti-oxidant activity, this compound can oxidize various thiols. However there are no detailed studies in the literature about the thiol oxidase-like activity of this compound against biologically significant mono and di-thiols with respect to various pH conditions. Keeping in mind the scarcity of data in this area of organochalcogen chemistry, we report for the first time the kinetics of thiol oxidation by diphenyl diselenide, which was carried out in a commonly used phosphate buffer, not only at physiological pH, but also at a number of acidic values. The relative reactivities of the different thiols with diphenyl diselenide were independent of the pKa of the thiol group, such that at pH 7.4, cysteine and dithiothreitol were the most reactive, while 2,3-dimercapto-1-propanesulfonic acid and glutathione were weakly reactive and extremely low reactivity was observed with dimercaptosuccinic acid. Rate of oxidation was dependent on the pH of the incubation medium. The results obtained will help us in the design of rational strategies for the safe pharmacological use of diphenyl diselenide.

Waseem Hassan; Joao Batista Teixeira Rocha

2012-01-01

70

Surface functionalization of ZnO films by THIOL  

Science.gov (United States)

In the present study, we have investigated the surface functionalization of dodecanethiol on ZnO film surface grown at three different oxygen partial pressure using pulsed laser deposition (PLD) technique. The thiol funcitonalized ZnO films have been examined by X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), photoluminescence (PL) and vibrating sample magnetometer (VSM) measurement. XRD pattern show slight decrease in intensity of diffraction peak upon surface functionalization. The chemical bonding of ZnO with thiol has been confirmed by XPS measurement. The presence of S 2p3/2 peak centered about 163 eV suggests that proton has dissociated fromed thiol and form chemical bonding to the ZnO surface. PL measurements of thiol functionalized ZnO films show quenching of visible emission intensity relative to the unfunctionalized ZnO films. The quenching of visible emission suggests that thiol passivates surface defects via Zn-S bonding. Surface functionalization of ZnO films with thiol induces robust room temperature ferromagnetism as evidenced from VSM measurements.

Jayalakshmi, G.; Saravanan, K.; Balasubramanian, T.

2012-10-01

71

Redox Initiation of Bulk Thiol-Ene Polymerizations.  

UK PubMed Central (United Kingdom)

The unique formation-structure-property attributes and reaction behavior of the thiol-ene "click" reaction have been explored extensively for photochemically and thermally initiated reactions but have been much less explored for redox initiation. Therefore, the objective of this work is to characterize fully the impact of the initiation system, monomer structure, degree of functionalization, and inhibitor level on the redox-mediated thiol-ene polymerization rate and behavior. Moreover, this study confirms the ability of redox initiation to achieve full conversion of desired thiol-ene "click" products for small molecules in solution. For the multifunctional thiol-ene systems, polymerization rate was shown to be comparable to photo- and thermally initiated systems, but with the additional advantages of unlimited depth of cure and mild reaction conditions. Additionally, the network properties of the redox-initiated thiol-ene systems were on par with a photocured material formulated with identical monomers and radical initiating potential. Lastly, control over the polymerization rate and preceding induction period was garnered from the concentration of inhibitor included in the reaction mixture. The mechanism of action of quinone inhibition in redox-mediated thiol-ene polymerizations is shown to depend on both the presence of an aniline reducing agent and the concentration of inhibitor, with quinone concentrations in great excess of oxidizing agent concentrations actually leading to heightened polymerization rates when aniline is present.

Cole MA; Jankousky KC; Bowman CN

2013-02-01

72

Modeling of farnesyltransferase inhibition by some thiol and non-thiol peptidomimetic inhibitors using genetic neural networks and RDF approaches.  

UK PubMed Central (United Kingdom)

Inhibition of farnesyltransferase (FT) enzyme by a set of 78 thiol and non-thiol peptidomimetic inhibitors was successfully modeled by a genetic neural network (GNN) approach, using radial distribution function descriptors. A linear model was unable to successfully fit the whole data set; however, the optimum Bayesian regularized neural network model described about 87% inhibitory activity variance with a relevant predictive power measured by q2 values of leave-one-out and leave-group-out cross-validations of about 0.7. According to their activity levels, thiol and non-thiol inhibitors were well-distributed in a topological map, built with the inputs of the optimum non-linear predictor. Furthermore, descriptors in the GNN model suggested the occurrence of a strong dependence of FT inhibition on the molecular shape and size rather than on electronegativity or polarizability characteristics of the studied compounds.

González MP; Caballero J; Tundidor-Camba A; Helguera AM; Fernández M

2006-01-01

73

Cell-type specific requirements for thiol/disulfide exchange during HIV-1 entry and infection  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background The role of disulfide bond remodeling in HIV-1 infection is well described, but the process still remains incompletely characterized. At present, the data have been predominantly obtained using established cell lines and/or CXCR4-tropic laboratory-adapted virus strains. There is also ambiguity about which disulfide isomerases/ reductases play a major role in HIV-1 entry, as protein disulfide isomerase (PDI) and/or thioredoxin (Trx) have emerged as the two enzymes most often implicated in this process. Results We have extended our previous findings and those of others by focusing on CCR5-using HIV-1 strains and their natural targets - primary human macrophages and CD4+ T lymphocytes. We found that the nonspecific thiol/disulfide exchange inhibitor, 5,5'-dithiobis(2-nitrobenzoic acid) (DTNB), significantly reduced HIV-1 entry and infection in cell lines, human monocyte-derived macrophages (MDM), and also phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC). Subsequent studies were performed using specific anti-PDI or Trx monoclonal antibodies (mAb) in HIV-1 envelope pseudotyped and wild type (wt) virus infection systems. Although human donor-to-donor variability was observed as expected, Trx appeared to play a greater role than PDI in HIV-1 infection of MDM. In contrast, PDI, but not Trx, was predominantly involved in HIV-1 entry and infection of the CD4+/CCR5+ T cell line, PM-1, and PHA-stimulated primary human T lymphocytes. Intriguingly, both PDI and Trx were present on the surface of MDM, PM-1 and PHA-stimulated CD4+ T cells. However, considerably lower levels of Trx were detected on freshly isolated CD4+ lymphocytes, compared to PHA-stimulated cells. Conclusions Our findings clearly demonstrate the role of thiol/disulfide exchange in HIV-1 entry in primary T lymphocytes and MDM. They also establish a cell-type specificity regarding the involvement of particular disulfide isomerases/reductases in this process and may provide an explanation for differences among previously published studies. More importantly, from an in vivo perspective, the preferential utilization of PDI may be relevant to the HIV-1 entry and establishment of virus reservoirs in resting CD4+ cells, while the elevated levels of Trx reported in the chronic stages of HIV-1 infection may facilitate the virus entry in macrophages and help to sustain high viremia during the decline of T lymphocytes.

Stantchev Tzanko S; Paciga Mark; Lankford Carla R; Schwartzkopff Franziska; Broder Christopher C; Clouse Kathleen A

2012-01-01

74

Efficient functionalization of oxide-free silicon(111) surfaces: thiol-yne versus thiol-ene click chemistry.  

UK PubMed Central (United Kingdom)

Thiol-yne click (TYC) chemistry was utilized as a copper-free click reaction for the modification of alkyne-terminated monolayers on oxide-free Si(111) surfaces, and the results were compared with the analogous thiol-ene click (TEC) chemistry. A wide range of thiols such as 9-fluorenylmethoxy-carbonyl cysteine, thio-?-d-glucose tetraacetate, thioacetic acid, thioglycerol, thioglycolic acid, and 1H,1H,2H,2H-perfluorodecanethiol was immobilized using TYC under photochemical conditions, and all modified surfaces were characterized by static water contact angle measurements, X-ray photoelectron spectroscopy (including a simulation thereof by density functional calculations), and infrared absorption reflection spectroscopy. Surface-bound TYC proceeds with an efficiency of up to 1.5 thiols per alkyne group. This high surface coverage proceeds without oxidizing the Si surface. TYC yielded consistently higher surface coverages than TEC, due to double addition of thiols to alkyne-terminated monolayers. This also allows for the sequential and highly efficient attachment of two different thiols onto an alkyne-terminated monolayer.

Bhairamadgi NS; Gangarapu S; Caipa Campos MA; Paulusse JM; van Rijn CJ; Zuilhof H

2013-04-01

75

Isolated menthone reductase and nucleic acid molecules encoding same  

Science.gov (United States)

The present invention provides isolated menthone reductase proteins, isolated nucleic acid molecules encoding menthone reductase proteins, methods for expressing and isolating menthone reductase proteins, and transgenic plants expressing elevated levels of menthone reductase protein.

Croteau, Rodney B; Davis, Edward M; Ringer, Kerry L

2013-04-23

76

Redox-dependent increases in glutathione reductase and exercise preconditioning: role of NADPH oxidase and mitochondria.  

UK PubMed Central (United Kingdom)

AIMS: We have previously shown that exercise leads to sustainable cardioprotection through a mechanism involving improved glutathione replenishment. This study was conducted to determine if redox-dependent modifications in glutathione reductase (GR) were involved in exercise cardioprotection. Furthermore, we sought to determine if reactive oxygen species generated by NADPH oxidase and/or mitochondria during exercise were triggering events for GR modulations. METHODS AND RESULTS: Rats were exercised for 10 consecutive days, after which isolated hearts were exposed to ischaemia/reperfusion (25 min/120 min). Exercise protected against infarction and arrhythmia, and preserved coronary flow. The GR inhibitor BCNU abolished the beneficial effects. GR activity was increased following exercise in a redox-dependent manner, with no change in GR protein levels. Because fluorescent labelling of GR protein thiols showed lower amounts of reduced thiols after exercise, we sought to determine the source of intracellular reactive oxygen species that may be activating GR. Subsets of animals were exercised immediately after treatment with either NADPH-oxidase inhibitors apocynin or Vas2870, or with mitoTEMPO or Bendavia, which reduce mitochondrial reactive oxygen species levels. The cardioprotective effects of exercise were abolished if animals exercised in the presence of NADPH oxidase inhibitors, in clear contrast to the mitochondrial reagents. These changes correlated with thiol-dependent modifications of GR. CONCLUSION: Adaptive cardioprotective signalling is triggered by reactive oxygen species from NADPH oxidase, and leads to improved glutathione replenishment through redox-dependent modifications in GR.

Frasier CR; Moukdar F; Patel HD; Sloan RC; Stewart LM; Alleman RJ; La Favor JD; Brown DA

2013-04-01

77

Designed Chemical Intervention with Thiols for Prophylactic Contraception  

Science.gov (United States)

Unlike somatic cells, sperm have several-fold more available-thiols that are susceptible to redox-active agents. The present study explains the mechanism behind the instant sperm-immobilizing and trichomonacidal activities of pyrrolidinium pyrrolidine-1-carbodithioate (PPC), a novel thiol agent rationally created for prophylactic contraception by minor chemical modifications of some known thiol drugs. PPC, and its three derivatives (with potential active-site blocked by alkylation), were synthesized and evaluated against live human sperm and metronidazole-susceptible and resistant Trichomonas vaginalis, in vitro. Sperm hexokinase activity was evaluated by coupled enzyme assay. PPC irreversibly immobilized 100% human sperm in ?30 seconds and totally eliminated Trichomonas vaginalis more efficiently than nonoxynol-9 and metronidazole. It significantly inhibited (P<0.001) thiol-sensitive sperm hexokinase. However, the molecule completely lost all its biological activities once its thiol group was blocked by alkylation. PPC was subsequently formulated into a mucoadhesive vaginal film using GRaS excipients and evaluated for spermicidal and microbicidal activities (in vitro), and contraceptive efficacy in rabbits. PPC remained fully active in quick-dissolving, mucoadhesive vaginal-film formulation, and these PPC-films significantly reduced pregnancy and fertility rates in rabbits. The films released ?90% of PPC in simulated vaginal fluid (pH 4.2) at 37°C in 5 minutes, in vitro. We have thus discovered a common target (reactive thiols) on chiefly-anaerobic, redox-sensitive cells like sperm and Trichomonas, which is susceptible to designed chemical interference for prophylactic contraception. The active thiol in PPC inactivates sperm and Trichomonas via interference with crucial sulfhydryl-disulfide based reactions, e.g. hexokinase activation in human sperm. In comparison to non-specific surfactant action of OTC spermicide nonoxynol-9, the action of thiol-active PPC is apparently much more specific, potent and safe. PPC presents a proof-of-concept for prophylactic contraception via manipulation of thiols in vagina for selective targeting of sperm and Trichomonas, and qualifies as a promising lead for the development of dually protective vaginal-contraceptive.

Jain, Ashish; Verma, Vikas; Sharma, Vikas; Kushwaha, Bhavana; Lal, Nand; Kumar, Lalit; Rawat, Tara; Dwivedi, Anil K.; Maikhuri, Jagdamba P.; Sharma, Vishnu L.; Gupta, Gopal

2013-01-01

78

Thiol-based redox switches and gene regulation.  

Science.gov (United States)

Cysteine is notable among the universal, proteinogenic amino acids for its facile redox chemistry. Cysteine thiolates are readily modified by reactive oxygen species (ROS), reactive electrophilic species (RES), and reactive nitrogen species (RNS). Although thiol switches are commonly triggered by disulfide bond formation, they can also be controlled by S-thiolation, S-alkylation, or modification by RNS. Thiol-based switches are common in both prokaryotic and eukaryotic organisms and activate functions that detoxify reactive species and restore thiol homeostasis while repressing functions that would be deleterious if expressed under oxidizing conditions. Here, we provide an overview of the best-understood examples of thiol-based redox switches that affect gene expression. Intra- or intermolecular disulfide bond formation serves as a direct regulatory switch for several bacterial transcription factors (OxyR, OhrR/2-Cys, Spx, YodB, CrtJ, and CprK) and indirectly regulates others (the RsrA anti-? factor and RegB sensory histidine kinase). In eukaryotes, thiol-based switches control the yeast Yap1p transcription factor, the Nrf2/Keap1 electrophile and oxidative stress response, and the Chlamydomonas NAB1 translational repressor. Collectively, these regulators reveal a remarkable range of chemical modifications exploited by Cys residues to effect changes in gene expression. PMID:20626317

Antelmann, Haike; Helmann, John D

2010-10-28

79

Thiol-based redox switches and gene regulation.  

UK PubMed Central (United Kingdom)

Cysteine is notable among the universal, proteinogenic amino acids for its facile redox chemistry. Cysteine thiolates are readily modified by reactive oxygen species (ROS), reactive electrophilic species (RES), and reactive nitrogen species (RNS). Although thiol switches are commonly triggered by disulfide bond formation, they can also be controlled by S-thiolation, S-alkylation, or modification by RNS. Thiol-based switches are common in both prokaryotic and eukaryotic organisms and activate functions that detoxify reactive species and restore thiol homeostasis while repressing functions that would be deleterious if expressed under oxidizing conditions. Here, we provide an overview of the best-understood examples of thiol-based redox switches that affect gene expression. Intra- or intermolecular disulfide bond formation serves as a direct regulatory switch for several bacterial transcription factors (OxyR, OhrR/2-Cys, Spx, YodB, CrtJ, and CprK) and indirectly regulates others (the RsrA anti-? factor and RegB sensory histidine kinase). In eukaryotes, thiol-based switches control the yeast Yap1p transcription factor, the Nrf2/Keap1 electrophile and oxidative stress response, and the Chlamydomonas NAB1 translational repressor. Collectively, these regulators reveal a remarkable range of chemical modifications exploited by Cys residues to effect changes in gene expression.

Antelmann H; Helmann JD

2011-03-01

80

Thiol/disulfide redox states in signaling and sensing.  

Science.gov (United States)

Rapid advances in redox systems biology are creating new opportunities to understand complexities of human disease and contributions of environmental exposures. New understanding of thiol-disulfide systems have occurred during the past decade as a consequence of the discoveries that thiol and disulfide systems are maintained in kinetically controlled steady states displaced from thermodynamic equilibrium, that a widely distributed family of NADPH oxidases produces oxidants that function in cell signaling and that a family of peroxiredoxins utilize thioredoxin as a reductant to complement the well-studied glutathione antioxidant system for peroxide elimination and redox regulation. This review focuses on thiol/disulfide redox state in biologic systems and the knowledge base available to support development of integrated redox systems biology models to better understand the function and dysfunction of thiol-disulfide redox systems. In particular, central principles have emerged concerning redox compartmentalization and utility of thiol/disulfide redox measures as indicators of physiologic function. Advances in redox proteomics show that, in addition to functioning in protein active sites and cell signaling, cysteine residues also serve as redox sensors to integrate biologic functions. These advances provide a framework for translation of redox systems biology concepts to practical use in understanding and treating human disease. Biological responses to cadmium, a widespread environmental agent, are used to illustrate the utility of these advances to the understanding of complex pleiotropic toxicities. PMID:23356510

Go, Young-Mi; Jones, Dean P

2013-01-29

 
 
 
 
81

Thiol/disulfide redox states in signaling and sensing.  

UK PubMed Central (United Kingdom)

Rapid advances in redox systems biology are creating new opportunities to understand complexities of human disease and contributions of environmental exposures. New understanding of thiol-disulfide systems have occurred during the past decade as a consequence of the discoveries that thiol and disulfide systems are maintained in kinetically controlled steady states displaced from thermodynamic equilibrium, that a widely distributed family of NADPH oxidases produces oxidants that function in cell signaling and that a family of peroxiredoxins utilize thioredoxin as a reductant to complement the well-studied glutathione antioxidant system for peroxide elimination and redox regulation. This review focuses on thiol/disulfide redox state in biologic systems and the knowledge base available to support development of integrated redox systems biology models to better understand the function and dysfunction of thiol-disulfide redox systems. In particular, central principles have emerged concerning redox compartmentalization and utility of thiol/disulfide redox measures as indicators of physiologic function. Advances in redox proteomics show that, in addition to functioning in protein active sites and cell signaling, cysteine residues also serve as redox sensors to integrate biologic functions. These advances provide a framework for translation of redox systems biology concepts to practical use in understanding and treating human disease. Biological responses to cadmium, a widespread environmental agent, are used to illustrate the utility of these advances to the understanding of complex pleiotropic toxicities.

Go YM; Jones DP

2013-03-01

82

Thiol and non-thiol antioxidants effect radiation damage expressed by IEC-6 cells  

International Nuclear Information System (INIS)

Full text: The epithelial lining of the GI mucosal surface is traditionally viewed as a passive barrier serving a largely protective function. Recently, enterocytes have been seen to function as sensitive indicators of oxidative stress, with defined responses to shifts in the oxidative balance. Radiation damage, long recognized as the result of the generation of reactive oxygen species, would theoretically be modulated by the presence of radical scavenging anti-oxidants. Cultures of IEC-6 cells, a model for the gastrointestinal epithelium were found to have lower numbers of adherent cells in response either n-acetyl cysteine (NAC) or l-ascorbate in the medium. In both cases the response was dose dependent, with inhibition in response to l-ascorbate well established by 48 hours. However, in contrast to the thiol antioxidant NAC where a late recovery was observed at high dose, no statistically significant increase in adherent cell numbers were observed with high dose l-ascorbate, and adherent cell numbers actually fell off with time. Exposure to antioxidants potentiated the damage from x-ray irradiation further reducing cell numbers. While we have previously shown that this damage was not associated with mitotic delay or inhibition of proliferation, the mechanism of this response remains undefined. Non-adherent cells were found to increase with dose in the presence of antioxidants with those cells having morphology consistent with apoptotic cells including nuclear condensation, and blabbing. Annexin V cells increased in the non-adherent cell layer, but the numbers did not seem to account for the severe reduction in cell numbers observed. It has been suggested that a p53 mutation alters the response to oxidative damage in these cells via a thiol containing motif sensitive to the cellular glutathione pool resulting in an automatic signal to release from the basement membrane, however it does not explain the similarity in effects to ascorbate

2003-01-01

83

Organized thiol functional groups in mesoporous core shell colloids  

International Nuclear Information System (INIS)

The co-condensation in situ of tetraethoxysilane (TEOS) and mercaptopropyltrimethoxysilane (MPTMS) using cetyltrimethylammonium bromide (CTAB) as a template results in the synthesis of multilayered mesoporous structured SiO2 colloids with “onion-like” chemical environments. Thiol groups were anchored to an inner selected SiO2 porous layer in a bilayered core shell particle producing different chemical regions inside the colloidal layered structure. X-Ray Photoelectron Spectroscopy (XPS) shows a preferential anchoring of the –SH groups in the double layer shell system, while porosimetry and simple chemical modifications confirm that pores are accessible. We can envision the synthesis of interesting colloidal objects with defined chemical environments with highly controlled properties. - Graphical abstract: Mesoporous core shell SiO2 colloids with organized thiol groups. Highlights: ? Double shell mesoporous silica colloids templated with CTAB. ? Sequential deposition of mesoporous SiO2 layers with different chemistries. ? XPS shows the selective functionalization of mesoporous layers with thiol groups.

2012-01-01

84

Effect of ?-irradiation on thiol compounds in grapefruit  

International Nuclear Information System (INIS)

The effect of 60Co ?-irradiation on thiol compounds in grapefruit was investigated. Thiols were separated by HPLC and measured with a fluorescence detector. Reduced glutathione (GSH), cysteine (CySH), cysteinylglycine (CySGly), and a number of unknown peaks were observed in unirradiated grapefruit. GSH was the main thiol at an average concentration of 143.3 ?M. GSH content exponentially decreased with increased radiation doses, and after 100 krad only 80% of the original remained. The G value based on the result of 100 krad was 0.29. Authentic GSH in water or citrate buffer (pH 3) was converted mainly to its oxidized form (GSSG) with ?-irradiated grapefruits showed no equivalent increase, however.

1989-01-01

85

Process for the removal of thiols from hydrocarbon oils  

Energy Technology Data Exchange (ETDEWEB)

Thiol impurities are absorbed and removed from hydrocarbon oils by contacting the oil in the absence of molecular oxygen with a scavenger at a temperature in the range of about 120/sup 0/ to 400/sup 0/C. The scavenger is a composite having a copper component and an inorganic porous carrier component and having a surface area in the range 20 to 1000 square meters per gram. The contacting must be discontinued when the thiol impurity content of the effluent product exceeds about 0.3 ppM.

Gibson, K.R.; Jacobson, R.L.

1980-05-27

86

Bioreductive activation of quinones: redox properties and thiol reactivity.  

UK PubMed Central (United Kingdom)

Redox properties and thiol reactivity are central to the therapeutic and toxicological properties of quinones. The use of other physicochemical parameters to establish predictive relationships for redox properties of quinones is discussed, and attention drawn to situations where such relationships may be unreliable. The rates of reaction of semiquinone radicals with oxygen, including those of chemotherapeutic agents such as mitomycin and the anthracyclines, can be predicted with reasonable confidence from the redox properties. The reactions of quinones with thiols such as glutathione produces reduced quinones and radicals, but the reactions are complex and all the features are not well understood.

Wardman P

1990-01-01

87

Efficient curing of vinyl carbonates by thiol-ene polymerization.  

UK PubMed Central (United Kingdom)

Vinyl carbonates have recently been identified as a suitable alternative to (meth)acrylates, especially due to the low irritancy and cytotoxicity of these monomers. The drawback of some vinyl carbonates containing abstractable hydrogens arises through their moderate reactivity compared with acrylates. Within this paper, we use the thiol-ene concept to enhance the photoreactivity of vinyl carbonates to a large extent to reach the level of those of similar acrylates. Mechanical properties of the final thiol-ene polymers were determined by nanoindentation. Furthermore, low toxicity of all components was confirmed by osteoblast cell culture experiments.

Mautner A; Qin X; Kapeller B; Russmueller G; Koch T; Stampfl J; Liska R

2012-12-01

88

5-Furan-2yl[1,3,4]oxadiazole-2-thiol, 5-Furan-2yl-4H [1,2,4] triazole-3-thiol and Their Thiol-Thione Tautomerism  

Directory of Open Access Journals (Sweden)

Full Text Available 5-Furan-2-yl[1,3,4]oxadiazole-2-thiol (Ia) and 5-furan-2-yl-4H-[1,2,4]-triazole-3-thiol (Ib) were synthesized from furan-2-carboxylic acid hydrazide. Mannich basesand methyl derivatives were then prepared. The structures of the synthesized compoundswere confirmed by elemental analyses, IR and 1H-NMR spectra. Their thiol-thione tautomericequilibrium is described.

M. Koparır; A. Çetin; A. Cansız

2005-01-01

89

The binding sites on human heme oxygenase-1 for cytochrome p450 reductase and biliverdin reductase.  

Science.gov (United States)

Human heme oxygenase-1 (hHO-1) catalyzes the NADPH-cytochrome P450 reductase-dependent oxidation of heme to biliverdin, CO, and free iron. The biliverdin is subsequently reduced to bilirubin by biliverdin reductase. Earlier kinetic studies suggested that biliverdin reductase facilitates the release of biliverdin from hHO-1 (Liu, Y., and Ortiz de Montellano, P. R. (2000) J. Biol. Chem. 275, 5297-5307). We have investigated the binding of P450 reductase and biliverdin reductase to truncated, soluble hHO-1 by fluorescence resonance energy transfer and site-specific mutagenesis. P450 reductase and biliverdin reductase bind to truncated hHO-1 with Kd = 0.4 +/- 0.1 and 0.2 +/- 0.1 microm, respectively. FRET experiments indicate that biliverdin reductase and P450 reductase compete for binding to truncated hHO-1. Mutation of surface ionic residues shows that hHO-1 residues Lys18, Lys22, Lys179, Arg183, Arg198, Glu19, Glu127, and Glu190 contribute to the binding of cytochrome P450 reductase. The mutagenesis results and a computational analysis of the protein surfaces partially define the binding site for P450 reductase. An overlapping binding site including Lys18, Lys22, Lys179, Arg183, and Arg185 is similarly defined for biliverdin reductase. These results confirm the binding of biliverdin reductase to hHO-1 and define binding sites of the two reductases. PMID:12626517

Wang, Jinling; de Montellano, Paul R Ortiz

2003-03-06

90

The binding sites on human heme oxygenase-1 for cytochrome p450 reductase and biliverdin reductase.  

UK PubMed Central (United Kingdom)

Human heme oxygenase-1 (hHO-1) catalyzes the NADPH-cytochrome P450 reductase-dependent oxidation of heme to biliverdin, CO, and free iron. The biliverdin is subsequently reduced to bilirubin by biliverdin reductase. Earlier kinetic studies suggested that biliverdin reductase facilitates the release of biliverdin from hHO-1 (Liu, Y., and Ortiz de Montellano, P. R. (2000) J. Biol. Chem. 275, 5297-5307). We have investigated the binding of P450 reductase and biliverdin reductase to truncated, soluble hHO-1 by fluorescence resonance energy transfer and site-specific mutagenesis. P450 reductase and biliverdin reductase bind to truncated hHO-1 with Kd = 0.4 +/- 0.1 and 0.2 +/- 0.1 microm, respectively. FRET experiments indicate that biliverdin reductase and P450 reductase compete for binding to truncated hHO-1. Mutation of surface ionic residues shows that hHO-1 residues Lys18, Lys22, Lys179, Arg183, Arg198, Glu19, Glu127, and Glu190 contribute to the binding of cytochrome P450 reductase. The mutagenesis results and a computational analysis of the protein surfaces partially define the binding site for P450 reductase. An overlapping binding site including Lys18, Lys22, Lys179, Arg183, and Arg185 is similarly defined for biliverdin reductase. These results confirm the binding of biliverdin reductase to hHO-1 and define binding sites of the two reductases.

Wang J; de Montellano PR

2003-05-01

91

Overexpression of chloroplast NADPH-dependent thioredoxin reductase in Arabidopsis enhances leaf growth and elucidates in vivo function of reductase and thioredoxin domains  

Science.gov (United States)

Plant chloroplasts have versatile thioredoxin systems including two thioredoxin reductases and multiple types of thioredoxins. Plastid-localized NADPH-dependent thioredoxin reductase (NTRC) contains both reductase (NTRd) and thioredoxin (TRXd) domains in a single polypeptide and forms homodimers. To study the action of NTRC and NTRC domains in vivo, we have complemented the ntrc knockout line of Arabidopsis with the wild type and full-length NTRC genes, in which 2-Cys motifs either in NTRd, or in TRXd were inactivated. The ntrc line was also transformed either with the truncated NTRd or TRXd alone. Overexpression of wild-type NTRC promoted plant growth by increasing leaf size and biomass yield of the rosettes. Complementation of the ntrc line with the full-length NTRC gene containing an active reductase but an inactive TRXd, or vice versa, recovered wild-type chloroplast phenotype and, partly, rosette biomass production, indicating that the NTRC domains are capable of interacting with other chloroplast thioredoxin systems. Overexpression of truncated NTRd or TRXd in ntrc background did not restore wild-type phenotype. Modeling of the three-dimensional structure of the NTRC dimer indicates extensive interactions between the NTR domains and the TRX domains further stabilize the dimeric structure. The long linker region between the NTRd and TRXd, however, allows flexibility for the position of the TRXd in the dimer. Supplementation of the TRXd in the NTRC homodimer model by free chloroplast thioredoxins indicated that TRXf is the most likely partner to interact with NTRC. We propose that overexpression of NTRC promotes plant biomass yield both directly by stimulation of chloroplast biosynthetic and protective pathways controlled by NTRC and indirectly via free chloroplast thioredoxins. Our data indicate that overexpression of chloroplast thiol redox-regulator has a potential to increase biofuel yield in plant and algal species suitable for sustainable bioenergy production.

Toivola, Jouni; Nikkanen, Lauri; Dahlstrom, Kathe M.; Salminen, Tiina A.; Lepisto, Anna; Vignols, hb Florence; Rintamaki, Eevi

2013-01-01

92

A maize gene encoding an NADPH binding enzyme highly homologous to isoflavone reductases is activated in response to sulfur starvation.  

UK PubMed Central (United Kingdom)

we isolated a novel gene that is selectively induced both in roots and shoots in response to sulfur starvation. This gene encodes a cytosolic, monomeric protein of 33 kD that selectively binds NADPH. The predicted polypeptide is highly homologous ( > 70%) to leguminous isoflavone reductases (IFRs), but the maize protein (IRL for isoflavone reductase-like) belongs to a novel family of proteins present in a variety of plants. Anti-IRL antibodies specifically recognize IFR polypeptides, yet the maize protein is unable to use various isoflavonoids as substrates. IRL expression is correlated closely to glutathione availability: it is persistently induced in seedlings whose glutathione content is about fourfold lower than controls, and it is down-regulated rapidly when control levels of glutathione are restored. This glutathione-dependent regulation indicates that maize IRL may play a crucial role in the establishment of a thiol-independent response to oxidative stress under glutathione shortage conditions.

Petrucco S; Bolchi A; Foroni C; Percudani R; Rossi GL; Ottonello S

1996-01-01

93

A maize gene encoding an NADPH binding enzyme highly homologous to isoflavone reductases is activated in response to sulfur starvation.  

Science.gov (United States)

we isolated a novel gene that is selectively induced both in roots and shoots in response to sulfur starvation. This gene encodes a cytosolic, monomeric protein of 33 kD that selectively binds NADPH. The predicted polypeptide is highly homologous ( > 70%) to leguminous isoflavone reductases (IFRs), but the maize protein (IRL for isoflavone reductase-like) belongs to a novel family of proteins present in a variety of plants. Anti-IRL antibodies specifically recognize IFR polypeptides, yet the maize protein is unable to use various isoflavonoids as substrates. IRL expression is correlated closely to glutathione availability: it is persistently induced in seedlings whose glutathione content is about fourfold lower than controls, and it is down-regulated rapidly when control levels of glutathione are restored. This glutathione-dependent regulation indicates that maize IRL may play a crucial role in the establishment of a thiol-independent response to oxidative stress under glutathione shortage conditions. PMID:8597660

Petrucco, S; Bolchi, A; Foroni, C; Percudani, R; Rossi, G L; Ottonello, S

1996-01-01

94

Surface functionalized thiol-ene waveguides for ?uorescence biosensing in micro?uidic devices  

DEFF Research Database (Denmark)

Thiol-ene polymers possess physical, optical, and chemical characteristics thatmake them ideal substrates for the fabrication of optofluidic devices. In this work, thiol-ene polymers are used to simultaneously create microfluidic channels and optical waveguides in one simple moulding step. The reactive functional groups present at the surface of the thiol-ene polymer are subsequently used for the rapid, one step, site-specific functionalization of the waveguide with biological recognition molecules. It was found that while the bulk properties and chemical surface properties of thiol-ene materials vary considerably with variations in stoichiometric composition, their optical properties remain mostly unchanged with an average refractive index value of 1.566 ± 0.008 for thiol-ene substrates encompassing a range from 150% excess ene to 90% excess thiol. Microfluidic chips featuring thiol-ene waveguides were fabricated from 40% excess thiol thiol-ene to ensure the presence of thiol functional groups at the surface of the waveguide. Biotin alkyne was photografted at specific locations using a photomask, directly at the interface between the microfluidic channel and the thiol-ene waveguide prior to conjugation with fluorescently labeled streptavidin. Fluorescence excitation was achieved by launching light through the thiol-ene waveguide, revealing bright fluorescent patterns along the channel/waveguide interface

Feidenhans'l, Nikolaj Agentoft; Lafleur, Josiane P.

2013-01-01

95

Mercury-resistance and mercuric reductase activity in Chromobacterium, Erwinia, and Bacillus species  

Energy Technology Data Exchange (ETDEWEB)

Mercury resistant bacteria have been the most extensively studied of all the metal-tolerant bacteria. Mercury resistance is usually mediated by two distinctly different enzymes encoded by plasmids. Mercuric reductase reduces Hg/sup 2 +/ to metallic mercury (Hg/sup 0/). Organomercurial lyases have a molecular weight of 20,000 to 40,000, are composed of 1 or 2 subunits and require the presence of thiol. Plasmic-encoded Hg/sup 2 +/ resistance and mercuric reductase activity have not been detected in many species of bacteria. A Chromobacterium, Erwinia and Bacillus species isolated from environmental samples were capable of growth in the presence of 50 ..mu..M HgCl/sub 2/. Cell-free extracts of the 3 organisms exhibited mercuric reductase activity that oxidized NADPH in the presence of HgCl/sub 2/. Negligible oxidation of NADPH was observed in the absence of HgCl/sub 2/. The Chromobacterium sp. did not contain any plasmid DNA. This would suggest that Hg/sup 2 +/ resistance was carried on the chromosome in Chromobacterium. A single 3 Mdal plasmid in the Bacillus sp. was refractory to curing. The Erwinia sp. contained 3 plasmids which were also refractory to curing. The location of the resistance genes is unknown in the Bacillus and Erwinia isolates.

Trevors, J.T.

1987-06-01

96

Thiol methyltransferase activity in colonocytes and erythrocyte membranes.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

AIMS--To verify the improved thiol methyltransferase (TMT) assay and measure activity in isolated colonocytes and erythrocyte membranes of the same subjects. METHODS--High performance liquid chromatography with radioactivity detection was used to measure 14C-methylmercaptoethanol formation, the reac...

Babidge, W J; Millard, S H; Roediger, W E

97

Preparation of Novel Hydrolyzing Urethane Modified Thiol-Ene Networks  

Directory of Open Access Journals (Sweden)

Full Text Available Novel tetra-functional hydrolyzing monomers were prepared from the reaction of TEOS and select alkene-containing alcohols, ethylene glycol vinyl ether or 2-allyloxy ethanol, and combined with trimethylolpropane tris(3-mercaptopropionate) (tri-thiol) in a thiol-ene “click” polymerization reaction to produce clear, colorless thiol-ene networks using both radiation and thermal-cure techniques. These networks were characterized for various mechanical characteristics, and found to posses Tg’s (DSC), hardness, tack, and thermal stability (TGA) consistent with their molecular structures. A new ene-modified urethane oligomer was prepared based on the aliphatic polyisocyanate Desmodur® N 3600 and added to the thiol-ene hydrolyzable network series in increasing amounts, creating a phase-segregated material having two Tg’s. An increase in water absorption in the ene-modified urethane formulations leading to a simultaneous increase in the rate of hydrolysis was supported by TGA data, film hardness measurements, and an NMR study of closely related networks. This phenomenon was attributed to the additional hydrogen bonding elements and polar functionality brought to the film with the addition of the urethane segment. SEM was utilized for visual analysis of topographical changes in the film’s surface upon hydrolysis and provides support for surface-driven erosion. Coatings prepared in this study are intended for use as hydrolyzing networks for marine coatings to protect against ship fouling.

Nicole M. Mackey; Bridget S. Confait; James H. Wynne; J. Paige Buchanan

2011-01-01

98

Peptidyl diazomethyl ketones are specific inactivators of thiol proteinases  

Energy Technology Data Exchange (ETDEWEB)

Peptidyl diazomethyl ketones are specific inactivators of thiol proteinases being unreactive toward other classes of proteinases. Variation of the peptide portion of the reagents has provided affinity labels for the selective inactivation of the thiol endopeptidase cathepsin B, streptococcal proteinase, and clostripain and for the aminodipeptidase cathepsin C. Comparison of rates of inactivation of cathepsin B and streptococcal proteinase revealed a similarity in specificity, both enzymes showing a preference for a phenylalanine residue in the penultimate position of the peptide portion of the reagent. Reagents not satisfying the specificity of a particular thiol proteinase either did not inactivate or did so only very slowly. In some cases, the reagents bound to the enzyme in a manner unproductive for alkylation but productive for substrate behavior leading to destruction of the inhibitor by cleavage of the peptide portion. Thus, benzyloxycarbonyl-Phe-Gly-Phe diazomethyl ketone was rapidly cleaved by cathepsin B into benzyloxycarbonyl-Phe-Gly and phenylalanine diazomethyl ketone. Clostripain, an enzyme of trypsin-like specificity, was selectively inactivated by benzyloxycarbpnyl-Lys diazomethyl ketone at micromolar concentrations, but was inactivated extremely slowly by 2.5 x 10/sup -4/ M benzyloxycarbonyl-Phe-Ala diazomethyl ketone, a very effective inactivated by Gly-Phe diazomethyl ketone in the range of 10/sup -7/ to 10/sup -8/ M. By comparison, peptidyl diazomethyl ketones with blocked NH/sub 2/ terminals were considerably less effective. Peptidyl diazomethyl ketones are unreactive with such thiols as mercaptoethanol and glutathione.

Green, G.D.J.; Shaw, E.

1981-02-25

99

Thiol-inducible direct fluorescence monitoring of drug release.  

UK PubMed Central (United Kingdom)

A new bifunctional compound NCC, which undergoes thiol-mediated disulfide cleavage after cell entry, produces a red-shifted fluorescent emission in the cytosol and releases free active DNA alkylating agent CLB into the nucleus, and finally leads to DNA damage and cell death.

Wu J; Huang R; Wang C; Liu W; Wang J; Weng X; Tian T; Zhou X

2013-01-01

100

Oligomerization of Indole Derivatives with Incorporation of Thiols  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Abstract: Two molecules of indole derivative, e.g. indole-5-carboxylic acid, reacted with one molecule of thiol, e.g. 1,2-ethanedithiol, in the presence of trifluoroacetic acid to yield adducts such as 3-[2-(2-amino-5-carboxyphenyl)-1-(2-mercaptoethylthio)ethyl]-1Hindole-5-carboxylic acid. Parallel ...

Felikss Mutulis; Adolf Gogoll; Ilze Mutule; Sviatlana Yahorava; Aleh Yahorau; Edvards Liepinsh; Jarl E.S. Wikberg

 
 
 
 
101

Production of dithioselenides from thiols and selenium dioxide  

International Nuclear Information System (INIS)

The authors have established that the slow addition of a methanol solution of selenium dioxide to solutions of thiols in dioxane (molar ratio 2:1) leads to the formation of the corresponding thioselenides without side compounds. Under the influence of bases, light, or heat above 1500C these compounds eliminate amorphous selenium and are converted quantitatively into the known disulfides.

1986-01-01

102

Glutation. The thiol in intracellular is a good radioprotector?  

International Nuclear Information System (INIS)

[en] Since 1988 there is a changing opinion about the Glutation's role in intracellular radiation protection. We review the thiols mechanism of radiation protection with special attention to GSH; and present the new features about this topic. (Author) 125 ref

1994-01-01

103

Expression of increased immunogenicity by thiol-releasing tumor variants.  

UK PubMed Central (United Kingdom)

Even moderate variations of the extracellular cysteine concentration were previously shown to affect T cell functions in vitro despite high concentrations of cystine. We therefore analyzed the membrane transport activities of T cells for cysteine and cystine, and the role of low molecular weight thiol in T cell-mediated host responses against a T cell tumor in vivo. A series of T cell clones and tumors including the highly malignant lymphoma L5178Y ESb and its strongly immunogenic variant ESb-D was found to express extremely weak transport activity for cystine but strong transport activity for cysteine. However, not all cells showed the expected requirement for cysteine (or 2-mercaptoethanol (2-ME)) in the culture medium. One group of clones and tumors including the malignant ESb-lymphoma did not respond to changes of extracellular cystine concentrations and was strongly thiol dependent. This group released only little acid soluble thiol (cysteine) if grown in cystine-containing cultures. The other T cell lines, in contrast, were able to maintain high intracellular GSH levels and DNA synthesis activity in cystine-containing culture medium without cystein or 2-ME and released substantial amounts of thiol. This group included the immunogenic ESb-D line. Additional thiol-releasing ESb variants were obtained by culturing large numbers of L5178Y ESb tumor cells in cultures without cysteine or 2-ME. All of these ESb variants showed a significantly decreased tumorigenicity and some of them induced cytotoxic and protective host responses even against the malignant ESb parent tumor. Taken together, our experiments suggest that the host response against a tumor may be limited in certain cases by the failure of the stimulator (i.e., the tumor) cell to deliver sufficient amounts of cysteine to the responding T cells.

Lim JS; Eck HP; Gmünder H; Dröge W

1992-04-01

104

Neutron-gamma irradiation and protein thiols: development of a protein thiol evaluation micro-method and application to irradiated baboons  

International Nuclear Information System (INIS)

[en] The essential non-protein sulfhydryl compound implicated in cellular radioprotection is glutathione. Protein thiols seem to be also involved in this protection and might be scavengers for free radical injury. We developed an analytical procedure for protein thiols measurement and we applied this method in neutron-gamma irradiated baboons. Our results demonstrated the reliability and sensitivity of the procedure. They also a drastic decrease of in vivo protein thiols after irradiation. (author)

1994-01-01

105

Rapid photochemical surface patterning of proteins in thiol-ene based microfluidic devices  

DEFF Research Database (Denmark)

The suitable optical properties of thiol–ene polymers combined with the ease of modifying their surface for the attachment of recognition molecules make them ideal candidates in many biochip applications. This paper reports the rapid one-step photochemical surface patterning of biomolecules in microfluidic thiol–ene chips. This work focuses on thiol–ene substrates featuring an excess of thiol groups at their surface. The thiol–ene stoichiometric composition can be varied to precisely control the number of surface thiol groups available for surface modification up to an average surface density of 136 ! 17 SH nm"2. Biotin alkyne was patterned directly inside thiol–ene microchannels prior to conjugation with fluorescently labelled streptavidin. The surface bound conjugates were detected by evanescent waveinduced fluorescence (EWIF), demonstrating the success of the grafting procedure and its potential for biochip applications.

Lafleur, Josiane P.; Kwapiszewski, Radoslaw

2013-01-01

106

Stabilising cysteinyl thiol oxidation and nitrosation for proteomic analysis.  

UK PubMed Central (United Kingdom)

Oxidation and S-nitrosylation of cysteinyl thiols (Cys-SH) to sulfenic (Cys-SOH), sulfinic (Cys-SO2H), sulfonic acids (Cys-SO3H), disulphides and S-nitrosothiols are suggested as important post-translational modification that can activate or deactivate the function of many proteins. Non-enzymatic post-translational modifications to cysteinyl thiols have been implicated in a wide variety of physiological and pathophysiological states but have been difficult to monitor in a physiological setting because of a lack of experimental tools. The purpose of this review is to bring together the approaches that have been developed for stably trapping cysteine either in its reduced or oxidised forms for enrichment and or subsequent mass spectrometric analysis. These tools are providing insight into potential targets for post-translational modifications to cysteine modification in vivo.

Ratnayake S; Dias IH; Lattman E; Griffiths HR

2013-06-01

107

Contrasting bonding behavior of thiol molecules on carbon fullerene structures  

International Nuclear Information System (INIS)

We have performed semiempirical as well as ab initio density-functional theory (DFT) calculations at T=0 to analyze the equilibrium configurations and electronic properties of spheroidal C60 as well as of cylindrical armchair (5,5) and (8,8) fullerenes passivated with SCH3 and S(CH2)2CH3 thiols. Our structural results reveal that the lowest-energy configurations of the adsorbates strongly depend on their chain length and on the structure of the underlying substrate. In the low-coverage regime, both SCH3 and S(CH2)2CH3 molecules prefer to organize into a molecular cluster on one side of the C60 surface, providing thus a less protective organic coating for the carbon structure. However, with increasing the number of adsorbed thiols, a transition to a more uniform distribution is obtained, which actually takes place for six and eight adsorbed molecules when using S(CH2)2CH3 and SCH3 chains, respectively. In contrast, for the tubelike arrangements at the low-coverage regime, a quasi-one-dimensional zigzag organization of the adsorbates along the tubes is always preferred. The sulfur-fullerene bond is considerably strong and is at the origin of outward and lateral displacements of the carbon atoms, leading to the stabilization of three-membered rings on the surface (spheroidal structures) as well as to sizable nonuniform radial deformations (cylindrical configurations). The electronic spectrum of our thiol-passivated fullerenes shows strong variations in the energy difference between the highest occupied and lowest unoccupied molecular orbitals as a function of the number and distribution of adsorbed thiols, opening thus the possibility to manipulate the transport properties of these compounds by means of selective adsorption mechanisms.

2003-01-01

108

Association of hydrogenase and dithionite reductase activities with the nitrite reductase of Desulfovibrio desulfuricans  

Energy Technology Data Exchange (ETDEWEB)

The membrane bound respiratory nitrite reductase from Desulfovibrio desulfuricans contains six c-type heme groups and catalyzes the six electron reduction of nitrite to ammonia. The purified enzyme required an excess of reducing equivalents for reduction relative to the amount of nitrite consumed in its reoxidation. The anomaly could be accounted for in terms of the presence of low levels of dithionite reductase and hydrogenase activity in the preparation. Dithionite reductase may be an alternate activity of nitrite reductase, whereas hydrogenase was shown to be a contaminant. The contaminating hydrogenase used nitrite reductase as electron acceptor in preference to cytochrome c/sub 3/ or benzyl viologen.

Steenkamp, D.J.; Peck, H.D. Jr.

1980-05-14

109

Trichomonas vaginalis: metronidazole and other nitroimidazole drugs are reduced by the flavin enzyme thioredoxin reductase and disrupt the cellular redox system. Implications for nitroimidazole toxicity and resistance.  

UK PubMed Central (United Kingdom)

Infections with the microaerophilic parasite Trichomonas vaginalis are treated with the 5-nitroimidazole drug metronidazole, which is also in use against Entamoeba histolytica, Giardia intestinalis and microaerophilic/anaerobic bacteria. Here we report that in T. vaginalis the flavin enzyme thioredoxin reductase displays nitroreductase activity with nitroimidazoles, including metronidazole, and with the nitrofuran drug furazolidone. Reactive metabolites of metronidazole and other nitroimidazoles form covalent adducts with several proteins that are known or assumed to be associated with thioredoxin-mediated redox regulation, including thioredoxin reductase itself, ribonucleotide reductase, thioredoxin peroxidase and cytosolic malate dehydrogenase. Disulphide reducing activity of thioredoxin reductase was greatly diminished in extracts of metronidazole-treated cells and intracellular non-protein thiol levels were sharply decreased. We generated a highly metronidazole-resistant cell line that displayed only minimal thioredoxin reductase activity, not due to diminished expression of the enzyme but due to the lack of its FAD cofactor. Reduction of free flavins, readily observed in metronidazole-susceptible cells, was also absent in the resistant cells. On the other hand, iron-depleted T. vaginalis cells, expressing only minimal amounts of PFOR and hydrogenosomal malate dehydrogenase, remained fully susceptible to metronidazole. Thus, taken together, our data suggest a flavin-based mechanism of metronidazole activation and thereby challenge the current model of hydrogenosomal activation of nitroimidazole drugs.

Leitsch D; Kolarich D; Binder M; Stadlmann J; Altmann F; Duchêne M

2009-04-01

110

Organized thiol functional groups in mesoporous core shell colloids  

Energy Technology Data Exchange (ETDEWEB)

The co-condensation in situ of tetraethoxysilane (TEOS) and mercaptopropyltrimethoxysilane (MPTMS) using cetyltrimethylammonium bromide (CTAB) as a template results in the synthesis of multilayered mesoporous structured SiO{sub 2} colloids with 'onion-like' chemical environments. Thiol groups were anchored to an inner selected SiO{sub 2} porous layer in a bilayered core shell particle producing different chemical regions inside the colloidal layered structure. X-Ray Photoelectron Spectroscopy (XPS) shows a preferential anchoring of the -SH groups in the double layer shell system, while porosimetry and simple chemical modifications confirm that pores are accessible. We can envision the synthesis of interesting colloidal objects with defined chemical environments with highly controlled properties. - Graphical abstract: Mesoporous core shell SiO{sub 2} colloids with organized thiol groups. Highlights: Black-Right-Pointing-Pointer Double shell mesoporous silica colloids templated with CTAB. Black-Right-Pointing-Pointer Sequential deposition of mesoporous SiO{sub 2} layers with different chemistries. Black-Right-Pointing-Pointer XPS shows the selective functionalization of mesoporous layers with thiol groups.

Marchena, Martin H. [Gerencia Quimica, Centro Atomico Constituyentes, Comision Nacional de Energia Atomica (CNEA), Avda. Gral. Paz 1499, B1650KNA Buenos Aires (Argentina); Granada, Mara [Centro Atomico Bariloche-CNEA, 8400 San Carlos de Bariloche (Argentina); Instituto Balseiro-Centro Atomico Bariloche-CNEA, San Carlos de Bariloche 8400 (Argentina); Bordoni, Andrea V. [Gerencia Quimica, Centro Atomico Constituyentes, Comision Nacional de Energia Atomica (CNEA), Avda. Gral. Paz 1499, B1650KNA Buenos Aires (Argentina); Joselevich, Maria [Asociacion Civil Expedicion Ciencia, Cabrera 4948, C1414BGP Buenos Aires (Argentina); Troiani, Horacio [Centro Atomico Bariloche-CNEA, 8400 San Carlos de Bariloche (Argentina); Instituto Balseiro-Centro Atomico Bariloche-CNEA, San Carlos de Bariloche 8400 (Argentina); Williams, Federico J. [DQIAQyF-INQUIMAE FCEN, Universidad de Buenos Aires, Ciudad Universitaria, Pabellon II, C1428EHA Buenos Aires (Argentina); Wolosiuk, Alejandro, E-mail: wolosiuk@cnea.gov.ar [Gerencia Quimica, Centro Atomico Constituyentes, Comision Nacional de Energia Atomica (CNEA), Avda. Gral. Paz 1499, B1650KNA Buenos Aires (Argentina)

2012-03-15

111

Identification of the thiol ester lipids in apolipoprotein B  

International Nuclear Information System (INIS)

[en] Human plasma low-density lipoproteins of 1.032-1.043 g/mL density were totally delipidized. The reduced and carboxymethylated apolipoprotein B was incubated with 50 mM [14C] methylamine at pH 8.5 at 30 0C. Covalent incorporation of [14C] methylamine was observed with concomitant generation of new sulfhydryl groups, which could be blocked with [3H]- or [14C]iodoacetic acid. One type of the [14C] methylamine-modified products was separated from the protein and was found to be lipid in nature. Its R/sub f/ on thin-layer chromatography (TLC) was similar to that of the synthetic N-methyl fatty acyl amides. After purification with TLC and transesterification in 3 N methanolic HCl, methyl esters of C16 and C18 fatty acids at 1:1 ratio were identified by gas-liquid chromatography. The transesterification method was verified with the known N-methyl fatty acyl amides. These results suggest the presence of labile thiol ester linked palmitate and stearate in apolipoprotein B. Under mild alkaline conditions, the thiol ester bonds are broken by methylamine and form N-methyl fatty acyl amides and release new -SH groups. Intramolecular thiol ester bonds linked between cysteine side chains and acidic amino acid residues were also found present, which will be reported separately

1988-03-08

112

Mechanistic considerations for formaldehyde-induced bronchoconstriction involving S-nitrosoglutathione reductase.  

UK PubMed Central (United Kingdom)

Inhalation of formaldehyde vapor has long been suspected of producing airway pathophysiology such as asthma and hyperresponsivity, presumably via irritant mechanisms. Recent studies on asthma and airway biology implicate changes in nitric oxide (NO) disposition in the adverse effects of formaldehyde, principally because enzymatic reduction of the endogenous bronchodilator S-nitrosoglutathione (GSNO) is dependent upon GSNO reductase (formally designated as alcohol dehydrogenase-3, ADH3), which also serves as the primary enzyme for cellular detoxification of formaldehyde. Considering recent evidence that regulation of bronchodilators like GSNO might play a more important role in asthma than inflammation per se, formaldehyde also needs to be considered as influencing ADH3-mediated GSNO catabolism. This is due to changes in ADH3 cofactors and thiol redox state among several potential mechanisms. Data suggest that deregulation of GSNO turnover provides a plausible, enzymatically based mechanism by which formaldehyde might exacerbate asthma and induce bronchoconstriction.

Thompson CM; Grafström RC

2008-01-01

113

FRUCTOSE-6-PHOSPHATE REDUCTASE FROM SALMONELLA GALLINARUM  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Zancan, Glaci T. (Universidade do Paraná, Curitiba, Paraná, Brazil), and Metry Bacila. Fructose-6-phosphate reductase from Salmonella gallinarum. J. Bacteriol. 87:614–618. 1964.—A fructose-6-phosphate reductase present in cell-free extracts of Salmonella gallinarum was purified approximately 42 time...

Zancan, Glaci T.; Bacila, Metry

114

Aqueous and vapor phase mercury sorption by inorganic oxide materials functionalized with thiols and poly-thiols  

Energy Technology Data Exchange (ETDEWEB)

The objective of the study is the development of sorbents where the sorption sites are highly accessible for the capture of mercury from aqueous and vapor streams. Only a small fraction of the equilibrium capacity is utilized for a sorbent in applications involving short residence times (e.g., vapor phase capture of mercury from coal-fired power plant flue gases). So, dynamic capacity rather than equilibrium capacity is more relevant for these kinds of situations. Rapid sorption rates and higher dynamic capacity can be achieved by increasing the accessibility of active sites and decreasing the diffusional resistance to mass transport for the adsorbing species. This requires the use of open structured sorbent materials and attachment of functional groups on the external surface area of supports. The strong interaction of sulfur containing ligands (e.g., thiol) with mercury makes them suitable candidates for immobilization on these types of materials. In this study, inorganic oxide supports like alumina and silica are functionalized with thiol moieties like mercapto silane, cysteine and poly-cysteine for capturing mercury from aqueous and vapor phase. Aqueous phase Hg (II) sorption studies with cysteine/poly-cysteine functionalized silica showed that high dynamic capacity can be achieved by attaching active sites (thiol) on the external area of supports. Vapor phase Hg capture studies with thiol-functionalized mesoporous silica (Hg{sup 0} concentration = 3.37 mg/m{sup 3} with N{sub 2} as the carrier, gas temperature = 70 C) yielded a capacity of 143 {mu}g Hg/g for the sorbent. Although the sulfur content for the sorbent was low (0.80 wt. %) the molar ratio of Hg captured to sulfur was comparatively high (2.86 x 10{sup -3}) pointing to the high accessibility of sulfur sites. (orig.)

Makkuni, A.; Bachas, L.G.; Bhattacharyya, D. [University of Kentucky, Dept. of Chemical and Materials Engineering, Lexington, KY (United States); Varma, R.S.; Sikdar, S.K. [US EPA, Cincinnati, OH (United States)

2005-05-01

115

A fluorescent probe which allows highly specific thiol labeling at low pH  

DEFF Research Database (Denmark)

Determination of the thiol-disulfide status in biological systems is challenging as redox pools are easily perturbed during sample preparation. This is particularly pertinent under neutral to mildly alkaline conditions typically required for alkylation of thiols. Here we describe the synthesis and properties of a thiol-specific reagent, fluorescent cyclic activated disulfide (FCAD), which includes the fluorescein moiety as fluorophore and utilizes a variation of thiol-disulfide exchange chemistry. The leaving-group character of FCAD makes it reactive at pH 3, allowing modification at low pH, limiting thiol-disulfide exchange. Different applications are demonstrated including picomolar thiol detection, determination of redox potentials, and in-gel detection of labeled proteins.

Nielsen, Jonas W.; Jensen, Kristine Steen

2012-01-01

116

Neutron-gamma irradiation and protein thiols: development of a protein thiol evaluation micro-method and application to irradiated baboons; Irradiation neutron-gamma et groupements thiols proteiques: developpement d`une micromethode d`evaluation des thiols proteiques et application au babouin irradie  

Energy Technology Data Exchange (ETDEWEB)

The essential non-protein sulfhydryl compound implicated in cellular radioprotection is glutathione. Protein thiols seem to be also involved in this protection and might be scavengers for free radical injury. We developed an analytical procedure for protein thiols measurement and we applied this method in neutron-gamma irradiated baboons. Our results demonstrated the reliability and sensitivity of the procedure. They also a drastic decrease of in vivo protein thiols after irradiation. (author). 5 refs.

Chancerelle, Y.; Lafond, J.L.; Della-Maura, L.; Faure, P.; Mathieu, J.; Costa, P.; Mestries, J.C.; Kergonou, J.F.

1994-12-31

117

Fabrication and bonding of thiol-ene-based microfluidic devices : Technical Note  

DEFF Research Database (Denmark)

In this work, the bonding strength of microchips fabricated by thiol-ene free-radical polymerization was characterized in detail by varying the monomeric thiol/allyl composition from the stoichiometric ratio (1:1) up to 100% excess of thiol (2:1) or allyl (1:2) functional groups. Four different thiol-ene to thiol-ene bonding combinations were tested by bonding: (i) two stoichiometric layers, (ii) two layers bearing complementary excess of thiols and allyls, (iii) two layers both bearing excess of thiols, or (iv) two layers both bearing excess of allyls. The results showed that the stiffness of the cross-linked polymer plays the most crucial role regarding the bonding strength. The most rigid polymer layers were obtained by using the stoichiometric composition or an excess of allyls, and thus, the bonding combinations (i) and (iv) withstood the highest pressures (up to the cut-off value of 6 bar). On the other hand, excess of thiol monomers yielded more elastic polymer layers and thus decreased the pressure tolerance for bonding combinations (ii) and (iii). By using monomers with more thiol groups (e.g. tetrathiol versus trithiol), a higher cross-linking ratio, and thus, greater stiffness was obtained. Surface characterization by infrared spectroscopy confirmed that the changes in the monomeric thiol/allyl composition were also reflected in the surface chemistry. The flexibility of being able to bond different types of thiol-enes together allows for tuning of the surface chemistry to yield the desired properties for each application. Here, a capillary electrophoresis separation is performed to demonstrate the attractive properties of stoichiometric thiol-ene microchips.

Sikanen, Tiina M; Lafleur, Josiane P.

2013-01-01

118

Combined bead polymerization and Cinchona organocatalyst immobilization by thiol–ene addition  

Directory of Open Access Journals (Sweden)

Full Text Available In this work, we report an unusually concise immobilization of Cinchona organocatalysts using thiol–ene chemistry, in which catalyst immobilization and bead polymerization is combined in a single step. A solution of azo initiator, polyfunctional thiol, polyfunctional alkene and an unmodified Cinchona-derived organocatalyst in a solvent is suspended in water and copolymerized on heating by thiol–ene additions. The resultant spherical and gel-type polymer beads have been evaluated as organocatalysts in catalytic asymmetric transformations.

Kim A. Fredriksen; Tor E. Kristensen; Tore Hansen

2012-01-01

119

Appel-reagent-mediated transformation of glycosyl hemiacetal derivatives into thioglycosides and glycosyl thiols.  

Science.gov (United States)

A series of glycosyl hemiacetal derivatives have been transformed into thioglycosides and glycosyl thiols in a one-pot two-step reaction sequence mediated by Appel reagent (carbon tetrabromide and triphenylphosphine). 1,2-trans-Thioglycosides and ?-glycosyl thiol derivatives were stereoselectively formed by the reaction of the in situ generated glycosyl bromides with thiols and sodium carbonotrithioate. The reaction conditions are reasonably simple and yields were very good. PMID:23766814

Ghosh, Tamashree; Santra, Abhishek; Misra, Anup Kumar

2013-05-22

120

Appel-reagent-mediated transformation of glycosyl hemiacetal derivatives into thioglycosides and glycosyl thiols.  

UK PubMed Central (United Kingdom)

A series of glycosyl hemiacetal derivatives have been transformed into thioglycosides and glycosyl thiols in a one-pot two-step reaction sequence mediated by Appel reagent (carbon tetrabromide and triphenylphosphine). 1,2-trans-Thioglycosides and ?-glycosyl thiol derivatives were stereoselectively formed by the reaction of the in situ generated glycosyl bromides with thiols and sodium carbonotrithioate. The reaction conditions are reasonably simple and yields were very good.

Ghosh T; Santra A; Misra AK

2013-01-01

 
 
 
 
121

Alkyl hydroperoxide reductase repair by Helicobacter pylori methionine sulfoxide reductase.  

UK PubMed Central (United Kingdom)

Protein exposure to oxidants such as HOCl leads to formation of methionine sulfoxide (MetSO) residues, which can be repaired by methionine sulfoxide reductase (Msr). An H. pylori msr strain was more sensitive to HOCl-mediated killing than the parent. Because of its abundance in H. pylori and its high methionine content, alkyl hydroperoxide reductase (AhpC) was hypothesized to be prone to methionine oxidation. AhpC was expressed as a recombinant protein in E.coli. AhpC activity was abolished by HOCl, while all six methionine residues of the enzyme were fully to partially oxidized. Upon incubation with a Msr repair mixture, AhpC activity was restored to non-oxidized levels and the MetSO residues were repaired to methionine, albeit to different degrees. The two most oxidized and then Msr-repaired methionine residues in AhpC, Met101 and Met133, were substituted with isoleucine residues by site-directed mutagenesis, either individually or together. Escherichia coli cells expressing variant versions were more sensitive to t-butyl hydroperoxide than cells expressing native protein, and purified AhpC variant proteins had 5-39% of native enzyme activity. Variant proteins were still able to oligomerize like the native version and CD spectra of variant proteins revealed no significant change in AhpC conformation, indicating the loss of activity in these variants was not related to major structural alterations. Our results suggest Met101 and Met133 residues are both important for AhpC catalytic activity and their integrity relies on a functional Msr.

Benoit SL; Bayyareddy K; Mahawar M; Sharp JS; Maier RJ

2013-10-01

122

Recent progress in luminescent and colorimetric chemosensors for detection of thiols.  

UK PubMed Central (United Kingdom)

In the past few decades, the development of optical probes for thiols has attracted great attention because of the biological importance of the thiol-containing molecules such as cysteine (Cys), homocysteine (Hcy), and glutathione (GSH). This tutorial review focuses on various thiol detection methods based on luminescent or colorimetric spectrophotometry published during the period 2010-2012. The discussion covers a diversity of sensing mechanisms such as Michael addition, cyclization with aldehydes, conjugate addition-cyclization, cleavage of sulfonamide and sulfonate esters, thiol-halogen nucleophilic substitution, disulfide exchange, native chemical ligation (NCL), metal complex-displace coordination, and nanomaterial-related and DNA-based chemosensors.

Jung HS; Chen X; Kim JS; Yoon J

2013-07-01

123

Recent progress in luminescent and colorimetric chemosensors for detection of thiols.  

Science.gov (United States)

In the past few decades, the development of optical probes for thiols has attracted great attention because of the biological importance of the thiol-containing molecules such as cysteine (Cys), homocysteine (Hcy), and glutathione (GSH). This tutorial review focuses on various thiol detection methods based on luminescent or colorimetric spectrophotometry published during the period 2010-2012. The discussion covers a diversity of sensing mechanisms such as Michael addition, cyclization with aldehydes, conjugate addition-cyclization, cleavage of sulfonamide and sulfonate esters, thiol-halogen nucleophilic substitution, disulfide exchange, native chemical ligation (NCL), metal complex-displace coordination, and nanomaterial-related and DNA-based chemosensors. PMID:23689799

Jung, Hyo Sung; Chen, Xiaoqiang; Kim, Jong Seung; Yoon, Juyoung

2013-07-21

124

Synthesis of Oligonucleotides Carrying Thiol Groups Using a Simple Reagent Derived from Threoninol  

Directory of Open Access Journals (Sweden)

Full Text Available Oligonucleotides carrying thiol groups are useful intermediates for a remarkable number of applications involving nucleic acids. In this study, DNA oligonucleotides carrying tert-butylsulfanyl (t-BuS) protected thiol groups have been prepared. A building block derived from threoninol has been developed to introduce a thiol group at any predetemined position of an oligonucleotide. The resulting thiolated oligonucleotides have been used for the preparation of oligonucleotide conjugates and for the functionalization of gold nanoparticles using the reactivity of the thiol groups.

Sonia Pérez-Rentero; Santiago Grijalvo; Rubén Ferreira; Ramon Eritja

2012-01-01

125

Crystal Growth of Thiol-Stabilized Gold Nanoparticles by Heat-Induced Coalescence  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract A monolayer of dodecanethiol-stabilized gold nanoparticles changed into two-dimensional and three-dimensional self-organized structures by annealing at 323 K. Subsequent crystal growth of gold nanoparticles occurred. Thiol molecules, although chemisorbed, form relatively unstable bonds with the gold surface; a few thiols desorbed from the surface and oxidized to disulfides at 323 K, because the interaction energy between thiol macromolecules is larger than that between a thiol and a nanoparticle. The gold nanoparticles approached each other and grew into large single or twinned crystals because of the van der Waals attraction and the heat generated by the exothermic formation of disulfides.

Moon SookYoung; Tanaka Shun-ichiro; Sekino Tohru

2010-01-01

126

The thiol pool in human plasma: the central contribution of albumin to redox processes.  

UK PubMed Central (United Kingdom)

The plasma compartment has particular features regarding the nature and concentration of low and high molecular weight thiols and oxidized derivatives. Plasma is relatively poor in thiol-based antioxidants; thiols are in lower concentrations than in cells and mostly oxidized. The different thiol-disulfide pairs are not in equilibrium and the steady state concentrations of total thiols as well as reduced versus oxidized ratios are maintained by kinetic barriers, including the rates of reactions and transport processes. The single thiol of human serum albumin (HSA-SH) is the most abundant plasma thiol. It is an important target for oxidants and electrophiles due to its reactivity with a wide variety of species and its relatively high concentration. A relatively stable sulfenic acid (HSA-SOH) can be formed in albumin exposed to oxidants. Plasma increases in mixed disulfides (HSA-SSR) or in sulfinic (HSA-SO2H) and sulfonic acids (HSA-SO3H) are associated with different pathologies and may constitute biomarkers of the antioxidant role of the albumin thiol. In this work we provide a critical review of the plasma thiol pool with a focus on human serum albumin.

Turell L; Radi R; Alvarez B

2013-06-01

127

Thiol-chromene click chemistry: a coumarin-based derivative and its use as regenerable thiol probe and in bioimaging applications.  

UK PubMed Central (United Kingdom)

The synthesis and characterization of a coumarin-chromene (8, 9-dihydro-2H-cyclopenta[b]pyrano[2,3-f]chromene-2,10(7aH)-dione) (1) derivative and its use for thiol chemosensing in water was reported. Experimental details showed 1 acts as a probe for the detection of thiols including cysteine (Cys), homocysteine (Hcy) and glutathione (GSH), whereas amino acids which do not contain thiols induced no changes in UV-vis spectra and fluorescence emission properties of 1. A possible detection mechanism is a nucleophilic attack of thiols to the ?,?-unsaturated ketone in 1 that resulted in a fluorescent coumarin derivative. Further studies showed that 1-thiol derivatives can be applied to the design of regenerative chemodosimeters for Cu(2+), Hg(2+) and Cd(2+) in water based on M(n+)-promoted desulfurization and recovery of 1. Furthermore, the optical properties of the probe and its Cys-addition product were theoretically studied. The ability of probe 1 to detect thiols in living cells (HepG2 cells) via an enhancement of the fluorescence was proved. Moreover, the applicability of 1 for the direct determination of biorelevant thiols in a complex matrix such as human plasma was also demonstrated.

Yang Y; Huo F; Yin C; Zheng A; Chao J; Li Y; Nie Z; Martínez-Máñez R; Liu D

2013-09-01

128

Tetrathionate reductase of Salmonella thyphimurium: a molybdenum containing enzyme  

International Nuclear Information System (INIS)

Use of radioactive molybdenum demonstrates that the tetrathionate reductase of Salmonella typhimurium is a molydenum containing enzyme. It is proposed that this enzyme shares with other molybdo-proteins, such as nitrate reductase, a common molybdenum containing cofactor the defect of which leads to the loss of the tetrathionate reductase and nitrate reductase activities

1986-04-29

129

Tetrathionate reductase of Salmonella thyphimurium: a molybdenum containing enzyme  

Energy Technology Data Exchange (ETDEWEB)

Use of radioactive molybdenum demonstrates that the tetrathionate reductase of Salmonella typhimurium is a molydenum containing enzyme. It is proposed that this enzyme shares with other molybdo-proteins, such as nitrate reductase, a common molybdenum containing cofactor the defect of which leads to the loss of the tetrathionate reductase and nitrate reductase activities.

Hinojosa-Leon, M.; Dubourdieu, M.; Sanchez-Crispin, J.A.; Chippaux, M.

1986-04-29

130

Transsulfuration pathway thiols and methylated arginines: the Hunter Community Study.  

UK PubMed Central (United Kingdom)

BACKGROUND: Serum homocysteine, when studied singly, has been reported to be positively associated both with the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine [ADMA, via inhibition of dimethylarginine dimethylaminohydrolase (DDAH) activity] and with symmetric dimethylarginine (SDMA). We investigated combined associations between transsulfuration pathway thiols, including homocysteine, and serum ADMA and SDMA concentrations at population level. METHODS: Data on clinical and demographic characteristics, medication exposure, C-reactive protein, serum ADMA and SDMA (LC-MS/MS), and thiols (homocysteine, cysteine, taurine, glutamylcysteine, total glutathione, and cysteinylglycine; capillary electrophoresis) were collected from a sample of the Hunter Community Study on human ageing [n = 498, median age (IQR) = 64 (60-70) years]. RESULTS: REGRESSION ANALYSIS SHOWED THAT: a) age (P = 0.001), gender (P = 0.03), lower estimated glomerular filtration rate (eGFR, P = 0.08), body mass index (P = 0.008), treatment with beta-blockers (P = 0.03), homocysteine (P = 0.02), and glutamylcysteine (P = 0.003) were independently associated with higher ADMA concentrations; and b) age (P = 0.001), absence of diabetes (P = 0.001), lower body mass index (P = 0.01), lower eGFR (P<0.001), cysteine (P = 0.007), and glutamylcysteine (P < 0.001) were independently associated with higher SDMA concentrations. No significant associations were observed between methylated arginines and either glutathione or taurine concentrations. CONCLUSIONS: After adjusting for clinical, demographic, biochemical, and pharmacological confounders the combined assessment of transsulfuration pathway thiols shows that glutamylcysteine has the strongest and positive independent associations with ADMA and SDMA. Whether this reflects a direct effect of glutamylcysteine on DDAH activity (for ADMA) and/or cationic amino acid transport requires further investigations.

Mangoni AA; Zinellu A; Carru C; Attia JR; McEvoy M

2013-01-01

131

Transsulfuration Pathway Thiols and Methylated Arginines: The Hunter Community Study  

Science.gov (United States)

Background Serum homocysteine, when studied singly, has been reported to be positively associated both with the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine [ADMA, via inhibition of dimethylarginine dimethylaminohydrolase (DDAH) activity] and with symmetric dimethylarginine (SDMA). We investigated combined associations between transsulfuration pathway thiols, including homocysteine, and serum ADMA and SDMA concentrations at population level. Methods Data on clinical and demographic characteristics, medication exposure, C-reactive protein, serum ADMA and SDMA (LC-MS/MS), and thiols (homocysteine, cysteine, taurine, glutamylcysteine, total glutathione, and cysteinylglycine; capillary electrophoresis) were collected from a sample of the Hunter Community Study on human ageing [n?=?498, median age (IQR)?=?64 (60–70) years]. Results Regression analysis showed that: a) age (P?=?0.001), gender (P?=?0.03), lower estimated glomerular filtration rate (eGFR, P?=?0.08), body mass index (P?=?0.008), treatment with beta-blockers (P?=?0.03), homocysteine (P?=?0.02), and glutamylcysteine (P?=?0.003) were independently associated with higher ADMA concentrations; and b) age (P?=?0.001), absence of diabetes (P?=?0.001), lower body mass index (P?=?0.01), lower eGFR (P<0.001), cysteine (P?=?0.007), and glutamylcysteine (P<0.001) were independently associated with higher SDMA concentrations. No significant associations were observed between methylated arginines and either glutathione or taurine concentrations. Conclusions After adjusting for clinical, demographic, biochemical, and pharmacological confounders the combined assessment of transsulfuration pathway thiols shows that glutamylcysteine has the strongest and positive independent associations with ADMA and SDMA. Whether this reflects a direct effect of glutamylcysteine on DDAH activity (for ADMA) and/or cationic amino acid transport requires further investigations.

Mangoni, Arduino A.; Zinellu, Angelo; Carru, Ciriaco; Attia, John R.; McEvoy, Mark

2013-01-01

132

Reactivity assessment of chalcones by a kinetic thiol assay.  

UK PubMed Central (United Kingdom)

The electrophilic nature of chalcones (1,3-diphenylprop-2-en-1-ones) and many other ?,?-unsaturated carbonyl compounds is crucial for their biological activity, which is often based on thiol-mediated regulation processes. To better predict their biological activity a simple screening assay for the assessment of the second-order rate constants (k(2)) in thia-Michael additions was developed. Hence, a clear structure-activity relationship of 16 differentially decorated hydroxy-alkoxychalcones upon addition of cysteamine could be established. Moreover, amongst other naturally occurring ?,?-unsaturated carbonyl compounds k(2) values for curcumin and cinnamaldehyde were gained while cinnamic acids or esters gave no or very slow reactions.

Amslinger S; Al-Rifai N; Winter K; Wörmann K; Scholz R; Baumeister P; Wild M

2013-01-01

133

Functional Conducting Polymers via Thiol-ene Chemistry  

Directory of Open Access Journals (Sweden)

Full Text Available We demonstrate here that thiol-ene chemistry can be used to provide side-chain functionalized monomers based on 3,4-propylenedioxythiophene (ProDOT) containing ionic, neutral, hydrophobic, and hydrophilic side chains. All reactions gave high yields and purification could generally be accomplished through precipitation. These monomers were polymerized either chemically or electro-chemically to give soluble materials or conductive films, respectively. This strategy provides for facile tuning of the solubility, film surface chemistry, and film morphology of this class of conducting polymers.

Kathleen E. Feldman; David C. Martin

2012-01-01

134

CDNA for human methylenetetrahydrofolate reductase  

UK PubMed Central (United Kingdom)

PCT No. PCT/CA95/00314 Sec. 371 Date Feb. 12, 1997 Sec. 102(e) Date Feb. 12, 1997 PCT Filed May 25, 1995 PCT Pub. No. WO95/33054 PCT Pub. Date Dec. 7, 1995The present invention relates to a cDNA probe for human methylenetetrahydrofolate reductase (MTHFR), and its uses. The probe of the present invention may be used for the identification of sequence abnormalities in patients with severe or mild MTHFR deficiency, including cardiovascular patients and patients with neurologic symptoms. A human MTHFR protein which hybridizes to the probe of the present invention may be used for therapy of MTHFR-deficiency patients by biochemical or pharmacological approaches.

ROZEN RIMA; GOYETTE PHILIPPE

135

Fluorescence detection of glutathione reductase activity based on deoxyribonucleic acid-templated silver nanoclusters.  

Science.gov (United States)

Fluorescent silver nanoclusters stabilized by DNA (DNA-AgNCs) exhibit distinct response rates to thiol and disulfide. Glutathione reductase can catalyze the reduction of the oxidized glutathione (GSSG) quickly to reduced glutathione (GSH) in the presence of ?-nicotinamide adenine dinucleotide 2'-phosphate reduced tetrasodium salt hydrate (NADPH). Consequently, DNA-AgNCs can serve as a new fluorescent platform for assaying the glutathione reductase (GR) activity. This newly proposed assay has a high sensitivity and a good selectivity toward GR. The GR activity can be detected in the range of 0.2-2.0 mU mL(-1) with a minimum detectable concentration of 0.2 mU mL(-1). Pepsin, lysozyme, trypsin, avidin, thrombin, myoglobin, and BSA have little effect on the fluorescence intensity of DNA-AgNCs. The GR activity assay is successfully used to monitor the inhibition of GR activity by a typical inhibitor 1,3-bis(2-chloroethyl)-1-nitrosourea. PMID:23790299

Zhu, Shuyun; Zhao, Xian-en; Zhang, Wei; Liu, Zhongyuan; Qi, Wenjing; Anjum, Saima; Xu, Guobao

2013-05-13

136

Fluorescence detection of glutathione reductase activity based on deoxyribonucleic acid-templated silver nanoclusters.  

UK PubMed Central (United Kingdom)

Fluorescent silver nanoclusters stabilized by DNA (DNA-AgNCs) exhibit distinct response rates to thiol and disulfide. Glutathione reductase can catalyze the reduction of the oxidized glutathione (GSSG) quickly to reduced glutathione (GSH) in the presence of ?-nicotinamide adenine dinucleotide 2'-phosphate reduced tetrasodium salt hydrate (NADPH). Consequently, DNA-AgNCs can serve as a new fluorescent platform for assaying the glutathione reductase (GR) activity. This newly proposed assay has a high sensitivity and a good selectivity toward GR. The GR activity can be detected in the range of 0.2-2.0 mU mL(-1) with a minimum detectable concentration of 0.2 mU mL(-1). Pepsin, lysozyme, trypsin, avidin, thrombin, myoglobin, and BSA have little effect on the fluorescence intensity of DNA-AgNCs. The GR activity assay is successfully used to monitor the inhibition of GR activity by a typical inhibitor 1,3-bis(2-chloroethyl)-1-nitrosourea.

Zhu S; Zhao XE; Zhang W; Liu Z; Qi W; Anjum S; Xu G

2013-07-01

137

Kinetics and mechanisms of thiol-disulfide exchange covering direct substitution and thiol oxidation-mediated pathways.  

UK PubMed Central (United Kingdom)

SIGNIFICANCE: Disulfides are important building blocks in the secondary and tertiary structures of proteins, serving as inter- and intra-subunit cross links. Disulfides are also the major products of thiol oxidation, a process that has primary roles in defense mechanisms against oxidative stress and in redox regulation of cell signaling. Although disulfides are relatively stable, their reduction, isomerisation, and interconversion as well as their production reactions are catalyzed by delicate enzyme machineries, providing a dynamic system in biology. Redox homeostasis, a thermodynamic parameter that determines which reactions can occur in cellular compartments, is also balanced by the thiol-disulfide pool. However, it is the kinetic properties of the reactions that best represent cell dynamics, because the partitioning of the possible reactions depends on kinetic parameters. CRITICAL ISSUES: This review is focused on the kinetics and mechanisms of thiol-disulfide substitution and redox reactions. It summarizes the challenges and advances that are associated with kinetic investigations in small molecular and enzymatic systems from a rigorous chemical perspective using biological examples. The most important parameters that influence reaction rates are discussed in detail. RECENT ADVANCES AND FUTURE DIRECTIONS: Kinetic studies of proteins are more challenging than small molecules, and quite often investigators are forced to sacrifice the rigor of the experimental approach to obtain the important kinetic and mechanistic information. However, recent technological advances allow a more comprehensive analysis of enzymatic systems via using the systematic kinetics apparatus that was developed for small molecule reactions, which is expected to provide further insight into the cell's machinery.

Nagy P

2013-05-01

138

Thiol-specific phosphorescent imaging in living cells with an azobis(2,2'-bipyridine)-bridged dinuclear iridium(III) complex.  

Science.gov (United States)

Based on an azo-thiol redox reaction, a non-emissive dinuclear iridium(III) complex showed off-on phosphorescent response toward thiols and was developed to image thiol levels in living cells. PMID:23381732

Li, Guanying; Chen, Yu; Wu, Jingheng; Ji, Liangnian; Chao, Hui

2013-03-11

139

Regulation of human methylenetetrahydrofolate reductase by phosphorylation  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Methylenetetrahydrofolate reductase (MTHFR) catalyzes the reduction of methylenetetrahydrofolate to methyltetrahydrofolate, the methyl donor for the conversion of homocysteine to methionine. Regulation of MTHFR activity is crucial for maintaining cellular concentrations of methionine and S-adenosylm...

Yamada, Kazuhiro; Strahler, John R.; Andrews, Philip C.; Matthews, Rowena G.

140

Purification and characterization of a pineapple crown leaf thiol protease.  

UK PubMed Central (United Kingdom)

A thiol protease was isolated and purified from the crown leaf of pineapple, Ananas comosus (L.) Merr. cv. Queen, by an immunoaffinity procedure. After the purification to electrophoretic homogeneity, the enzyme was characterized with respect to some of its physico-chemical and kinetic properties. The molecular weight of the protease (22.4-22.9 kDa), Km (97 microM) and kcat (8.8 s(-1)) for its esterolytic cleavage of the synthetic protease substrate N(alpha)-CBZ-L-lysine p-nitrophenyl ester, the concentration of its thiol activator L-cysteine required for half maximal activation A0.5 (9.9 microM), optimum pH (6.5) for its proteolytic action on azocasein, T(1/2) (60 degrees C) for inactivation by heating the enzyme (35.5 microg protein/mL) in citrate buffer pH 6.0 for 15 min, and SH-group content (0.98 mol/mol enzyme) were determined. Most of these physicochemical and kinetic properties were found to be similar to those of the already well-characterized stem bromelain (EC 3.4.22.32). Thus, the immunoaffinity purified crown leaf protease appeared to be closely related to stem bromelain.

Singh LR; Devi TP; Devi SK

2004-02-01

 
 
 
 
141

Liquid phase oxidation of alkyl sulfides and thiols by hydroperoxides  

Energy Technology Data Exchange (ETDEWEB)

In general, jet fuels deteriorate in quality with time. One of the significant undesirable changes is the formation of insoluble material which can plug nozzles and filters and coat heat exchanger surfaces. Deposit formation in fuels is triggered by autoxidation reactions and is closely associated with hydroperoxide concentration. If the available oxygen is low and the temperature raised, the hydroperoxide concentration will be limited by free radical decomposition. This regimen (low oxygen and increasing temperature) is similar to the environment found in an aircraft fuel system. The composition of deposits affords clues to the molecular species involved in deposit formation and the mechanism of formation. Hetero-atoms (oxygen, nitrogen and sulfur) and ash have been found to comprise up to 40% of such deposits. The sulfur content of these deposits has been found to vary from 1 to 9%. Sulfur (0.4% max. allowed) is the most abundant hetero-atom present in jet fuels. This paper is concerned with the reaction between a primary autoxidation product, a hydroperoxide, and organo sulfides and thiols. Specifically, the authors examine the tert-butyl hydroperoxide oxidation of hexyl sulfide and dodecyl thiol in deaerated benzene at 120/sup 0/C. The reactions were studied for time periods from 15 min to 180 min. Additionally, the authors have developed reaction conditions and an analytical method of high reproducibility which may be applicable to the study of other hydroperoxide oxidative processes.

Mushrush, G.W.; Hardy, D.R.; Eaton, H.G.; Hazlett, R.N.

1986-04-01

142

Thioredoxin glutathione reductase as a novel drug target: evidence from Schistosoma japonicum.  

UK PubMed Central (United Kingdom)

BACKGROUND: Schistosomiasis remains a major public health concern affecting billions of people around the world. Currently, praziquantel is the only drug of choice for treatment of human schistosomiasis. The emergence of drug resistance to praziquantel in schistosomes makes the development of novel drugs an urgent task. Thioredoxin glutathione reductase (TGR) enzymes in Schistosoma mansoni and some other platyhelminths have been identified as alternative targets. The present study was designed to confirm the existense and the potential value of TGR as a target for development of novel antischistosomal agents in Schistosoma japonicum, a platyhelminth endemic in Asia. METHODS AND FINDINGS: After cloning the S. japonicum TGR (SjTGR) gene, the recombinant SjTGR selenoprotein was purified and characterized in enzymatic assays as a multifunctional enzyme with thioredoxin reductase (TrxR), glutathione reductase (GR) and glutaredoxin (Grx) activities. Immunological and bioinformatic analyses confirmed that instead of having separate TrxR and GR proteins in mammalian, S. japonicum only encodes TGR, which performs the functions of both enzymes and plays a critical role in maintaining the redox balance in this parasite. These results were in good agreement with previous findings in Schistosoma mansoni and some other platyhelminths. Auranofin, a known inhibitor against TGR, caused fatal toxicity in S. japonicum adult worms in vitro and reduced worm and egg burdens in S. japonicum infected mice. CONCLUSIONS: Collectively, our study confirms that a multifunctional enzyme SjTGR selenoprotein, instead of separate TrxR and GR enzymes, exists in S. japonicum. Furthermore, TGR may be a potential target for development of novel agents against schistosomes. This assumption is strengthened by our demonstration that the SjTGR is an essential enzyme for maintaining the thiol-disulfide redox homeostasis of S. japonicum.

Song L; Li J; Xie S; Qian C; Wang J; Zhang W; Yin X; Hua Z; Yu C

2012-01-01

143

Indicator approach to develop a chemosensor for the colorimetric sensing of thiol-containing water and its application for the thiol detection in plasma.  

UK PubMed Central (United Kingdom)

A strategy for the determination of the presence of thiol-containing amino acids was successfully established by simply assembling copper chloride and xylenol orange (3,3'-bis[N,N-bis(carboxymethyl)aminomethyl]-o-cresolsulfonephthalein trisodium salt; XO) in a 1 : 1 molar ratio in quasi-physiological water solution (pH 6.0). The copper(II)-XO ensemble was highly selective for thiol species such as cysteine, homocysteine, and glutathione without interference from other amino acids and could quantitatively detect thiol in the range from 10 to 200 ?M with a linear relationship having an average molar absorbance constant of 6530 L mol(-1) cm(-1) in pure water. The whole recognition process for thiol gave rise to a rapid visual color change from purple-red to yellow which can be observed simultaneously with the naked-eye.

Huo FJ; Yang YT; Su J; Sun YQ; Yin CX; Yan XX

2011-05-01

144

The role of thiols in cellular response to radiation and drugs  

Energy Technology Data Exchange (ETDEWEB)

Cellular nonprotein thiols (NPSH) consist of glutathione (GSH) and other low molecular weight species such as cysteine, cysteamine, and coenzyme A. GSH is usually less than the total cellular NPSH, and with thiol reactive agents, such as diethyl maleate (DEM), its rate of depletion is in part dependent upon the cellular capacity for its resynthesis. If resynthesis is blocked by buthionine-S,R-sulfoximine(BSO), the NPSH, including GSH, is depleted more rapidly, Cellular thiol depletion by diamide, N-ethylmaleimide, and BSO may render oxygenated cells more sensitive to radiation. These cells may or may not show a reduction in the oxygen enhancement ratio (OER). Human A549 lung carcinoma cells depleted of their NPSH either by prolonged culture or by BSO treatment do not show a reduced OER but do show increased aerobic responses to radiation. Some nitroheterocyclic radiosensitizing drugs also deplete cellular thiols under aerobic conditions. Such reactivity may be the reason that they show anomalous radiation sensitization (i.e., better than predicted on the basis of electron affinity). Other nitrocompounds, such as misonidazole, are activated under hypoxic conditions to radical intermediates. When cellular thiols are depleted peroxide is formed. Under hypoxic conditions thiols are depleted because metabolically reduced intermediates react with GSH instead of oxygen. Thiol depletion, under hypoxic conditions, may be the reason that misonidazole and other nitrocompounds show an extra enhancement ratio with hypoxic cells. Thiol depletion by DEM or BSO alters the radiation response of hypoxic cells to misonidazole.

Biaglow, J.E.; Varnes, M.E.; Clark, E.P.; Epp, E.R.

1983-09-01

145

Manganese-catalyzed cross-coupling of thiols with aryl iodides.  

UK PubMed Central (United Kingdom)

A manganese-catalyzed cross-coupling reaction of thiols with aryl iodides, furnishing aryl thioethers in good to excellent yields has been reported; the system shows good functional group tolerance and enables the sterically demanding aryl iodides to couple with thiols.

Liu TJ; Yi CL; Chan CC; Lee CF

2013-05-01

146

Determination of acidity and nucleophilicity in thiols by reaction with monobromobimane and fluorescence detection.  

UK PubMed Central (United Kingdom)

A method based on the differential reactivity of thiol and thiolate with monobromobimane (mBBr) has been developed to measure nucleophilicity and acidity of protein and low-molecular-weight thiols. Nucleophilicity of the thiolate is measured as the pH-independent second-order rate constant of its reaction with mBBr. The ionization constants of the thiols are obtained through the pH dependence of either second-order rate constant or initial rate of reaction. For readily available thiols, the apparent second-order rate constant is measured at different pHs and then plotted and fitted to an appropriate pH function describing the observed number of ionization equilibria. For less available thiols, such as protein thiols, the initial rate of reaction is determined in a wide range of pHs and fitted to the appropriate pH function. The method presented here shows excellent sensitivity, allowing the use of nanomolar concentrations of reagents. The method is suitable for scaling and high-throughput screening. Example determinations of nucleophilicity and pK(a) are presented for captopril and cysteine as low-molecular-weight thiols and for human peroxiredoxin 5 and Trypanosoma brucei monothiol glutaredoxin 1 as protein thiols.

Sardi F; Manta B; Portillo-Ledesma S; Knoops B; Comini MA; Ferrer-Sueta G

2013-04-01

147

Combined bead polymerization and Cinchona organocatalyst immobilization by thiol–ene addition  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In this work, we report an unusually concise immobilization of Cinchona organocatalysts using thiol–ene chemistry, in which catalyst immobilization and bead polymerization is combined in a single step. A solution of azo initiator, polyfunctional thiol, polyfunctional alkene and an unmodified Cinchon...

Fredriksen, Kim A; Kristensen, Tor E; Hansen, Tore

148

Synthesis and Biological Evaluation of Novel Benzothiazole-2-thiol Derivatives as Potential Anticancer Agents  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A series of novel benzothiazole-2-thiol derivatives were synthesized and their structures determined by 1H-NMR, 13C-NMR and HRMS (ESI). The effects of all compounds on a panel of different types of human cancer cell lines were investigated. Among them, pyridinyl-2-amine linked benzothiazole-2-thiol ...

Xuan-Hong Shi; Zhao Wang; Yong Xia; Ting-Hong Ye; Mei Deng; You-Zhi Xu; Yu-Quan Wei; Luo-Ting Yu

149

Multiple aldehyde reductases of human brain.  

UK PubMed Central (United Kingdom)

Human brain contains four forms of aldehyde reducing enzymes. One major activity, designated AR3, has properties indicating its identity with the NADPH-dependent aldehyde reductase, EC 1.1.1.2. The other major form of human brain enzyme, AR1, which is also NADPH-dependent, reduces both aldehyde and ketone-containing substrates, including vitamin K3 (menadione) and daunorubicin, a cancer chemotherapeutic agent. This enzyme is very sensitive to inhibition by the flavonoids quercitrin and quercetine, and may be analogous to a daunorubicin reductase previously described in liver of other species. One minor form of human brain aldehyde reductase, AR2, demonstrates substrate specificity and inhibitor sensitivity which suggest its similarity to aldose reductases found in lens and other tissues of many species. This enzyme, which can also use NADH as cofactor to some extent, is the most active in reducing the aldehyde derivatives of the biogenic amines. The fourth human brain enzyme ("SSA reductase") differs from the other forms in its ability to use NADH as well as or better than NADPH as cofactor, and in its molecular weight, which is nearly twice that of the other forms. It is quite specific for succinic semialdehyde (SSA) as substrate, and was found to be significantly inhibited only by quercetine and quercitrin. AR3 can also reduce SSA, and both enzymes may contribute to the production of gamma-hydroxybutyric acid in vivo. These results indicate that the human brain aldehyde reductases can play relatively specific physiologic roles.

Hoffman PL; Wermuth B; von Wartburg JP

1980-01-01

150

Multiple aldehyde reductases of human brain.  

Science.gov (United States)

Human brain contains four forms of aldehyde reducing enzymes. One major activity, designated AR3, has properties indicating its identity with the NADPH-dependent aldehyde reductase, EC 1.1.1.2. The other major form of human brain enzyme, AR1, which is also NADPH-dependent, reduces both aldehyde and ketone-containing substrates, including vitamin K3 (menadione) and daunorubicin, a cancer chemotherapeutic agent. This enzyme is very sensitive to inhibition by the flavonoids quercitrin and quercetine, and may be analogous to a daunorubicin reductase previously described in liver of other species. One minor form of human brain aldehyde reductase, AR2, demonstrates substrate specificity and inhibitor sensitivity which suggest its similarity to aldose reductases found in lens and other tissues of many species. This enzyme, which can also use NADH as cofactor to some extent, is the most active in reducing the aldehyde derivatives of the biogenic amines. The fourth human brain enzyme ("SSA reductase") differs from the other forms in its ability to use NADH as well as or better than NADPH as cofactor, and in its molecular weight, which is nearly twice that of the other forms. It is quite specific for succinic semialdehyde (SSA) as substrate, and was found to be significantly inhibited only by quercetine and quercitrin. AR3 can also reduce SSA, and both enzymes may contribute to the production of gamma-hydroxybutyric acid in vivo. These results indicate that the human brain aldehyde reductases can play relatively specific physiologic roles. PMID:7424738

Hoffman, P L; Wermuth, B; von Wartburg, J P

1980-01-01

151

Effect of thiol compounds on bleomycin-induced DNA and chromosome damage in human cells.  

UK PubMed Central (United Kingdom)

Non-protein thiols are considered radioprotectors, preventing DNA damage by ionizing radiation. As bleomycin (BLM) is a radiomimetic agent it was proposed that thiols may prevent DNA damage produced by this antibiotic. However, results obtained with thiols and BLM-combined treatments in living cells are contradictory. The goal of this work was to assess the influence of five non-protein thiols of different electrical charge and chemical composition, on the DNA damage, DNA repair, chromosomal aberrations and cell killing induced by BLM. We found that, at the chromosomal level and cell killing, Glutathione, ?-Mercaptoethanol and cysteine showed a protective effect, while ditiothreitol and cysteamine increased them, whereas at the DNA level all thiols potentiated the DNA damage induced by BLM, most probably due to a reactivation of the BLM complex.

Mira A; Gimenez EM; Bolzán AD; Bianchi MS; López-Larraza DM

2013-01-01

152

Thiol-independent activity of a cholesterol-binding enterohemolysin produced by enteropathogenic Escherichia coli  

Directory of Open Access Journals (Sweden)

Full Text Available Enterohemolysin produced by Escherichia coli associated with infant diarrhea showed characteristics similar to those of thiol-activated hemolysins produced by Gram-positive bacteria, including inactivation by cholesterol, lytic activity towards eukaryotic cells and thermoinstability. However, enterohemolysin activity was not inactivated by oxidation or by SH group-blocking agents (1 mM HgCl2, 1 mM iodoacetic acid) and the hemolysin (100 µg/ml) was not lethal to mice, in contrast to the lethality of the thiol-activated hemolysin family to animals. Earlier reports showed that intravenous injection of partially purified streptolysin O preparations (0.2 µg) was rapidly lethal to mice. These results suggest that E. coli enterohemolysin is not a thiol-activated hemolysin, despite its ability to bind cholesterol, probably due to the absence of free thiol-group(s) that characterize the active form of the thiol-activated hemolysin molecule.

Figueirêdo P.M.S.; Catani C.F.; Yano T.

2003-01-01

153

Model studies for the thiol-mediated methyl transfer to corrinoids.  

Science.gov (United States)

The thiol-dependent methylation of heptamethyl cob(II)yrinate 8r with methyl iodide and methyl tosylate was explored under a variety of conditions. The interaction of the heptamethyl cob(II)yrinate with a variety of thiols was monitored prior to the addition of the methylating agent, and the formation of the Co(I) complex was only apparent in the reaction with hexane thiol. Nevertheless, thiol-mediated methylation of the Co(II) complex 8r takes place with methyl iodide under most conditions. The Co-methylation with methyl tosylate showed a different reactivity, was inhibited by pyridine or N-methylimidazole, and was strongly dependent on the the acidity of the thiol used. Mechanistic aspects are discussed. PMID:17581656

Wedemeyer-Exl, Christina; Darbre, Tamis; Keese, Reinhart

2007-05-25

154

Model studies for the thiol-mediated methyl transfer to corrinoids.  

UK PubMed Central (United Kingdom)

The thiol-dependent methylation of heptamethyl cob(II)yrinate 8r with methyl iodide and methyl tosylate was explored under a variety of conditions. The interaction of the heptamethyl cob(II)yrinate with a variety of thiols was monitored prior to the addition of the methylating agent, and the formation of the Co(I) complex was only apparent in the reaction with hexane thiol. Nevertheless, thiol-mediated methylation of the Co(II) complex 8r takes place with methyl iodide under most conditions. The Co-methylation with methyl tosylate showed a different reactivity, was inhibited by pyridine or N-methylimidazole, and was strongly dependent on the the acidity of the thiol used. Mechanistic aspects are discussed.

Wedemeyer-Exl C; Darbre T; Keese R

2007-07-01

155

A novel and rapid apoptosis assay based on thiol redox status.  

UK PubMed Central (United Kingdom)

We present here a novel probe, VitaBright-48, for the evaluation of the cellular level of reduced thiols. Using different cell lines and apoptogenic agents we show that a decrease in the level of reduced thiols correlates with well-known apoptotic markers such as phosphatidylserine translocation and caspase activity. The cell population to be investigated is added to the nonfluorescent stain VitaBright-48, which immediately permeates the cell membrane and reacts with intracellular thiols, forming a fluorescent compound. Quantification of the cell fluorescence directly after staining (without washing) can then be used to determine the population's cellular thiol level at the single cell level. Based on the results presented here, we suggest that measurement of changes in the level of free thiols should be added to the list of phenotypes which may be investigated in order to detect apoptosis.

Skindersoe ME; Rohde M; Kjaerulff S

2012-05-01

156

Towards thiol functionalization of vanadium pentoxide nanotubes using gold nanoparticles  

International Nuclear Information System (INIS)

Template-directed synthesis is a promising route to realize vanadate-based 1-D nanostructures, an example of which is the formation of vanadium pentoxide nanotubes and associated nanostructures. In this work, we report the interchange of long-chained alkyl amines with alkyl thiols. This reaction was followed using gold nanoparticles prepared by the Chemical Liquid Deposition (CLD) method with an average diameter of ?0.9nm and a stability of ?85 days. V2O5 nanotubes (VOx-NTs) with lengths of ?2?m and internal hollow diameters of 20-100nm were synthesized and functionalized in a Au-acetone colloid with a nominal concentration of ?4x10-3mol dm-3. The interchange reaction with dodecylamine is found only to occur in polar solvents and incorporation of the gold nanoparticles is not observed in the presence of n-decane.

2007-04-12

157

Reactivity assessment of chalcones by a kinetic thiol assay.  

Science.gov (United States)

The electrophilic nature of chalcones (1,3-diphenylprop-2-en-1-ones) and many other ?,?-unsaturated carbonyl compounds is crucial for their biological activity, which is often based on thiol-mediated regulation processes. To better predict their biological activity a simple screening assay for the assessment of the second-order rate constants (k(2)) in thia-Michael additions was developed. Hence, a clear structure-activity relationship of 16 differentially decorated hydroxy-alkoxychalcones upon addition of cysteamine could be established. Moreover, amongst other naturally occurring ?,?-unsaturated carbonyl compounds k(2) values for curcumin and cinnamaldehyde were gained while cinnamic acids or esters gave no or very slow reactions. PMID:23224077

Amslinger, Sabine; Al-Rifai, Nafisah; Winter, Katrin; Wörmann, Kilian; Scholz, Rebekka; Baumeister, Paul; Wild, Martin

2012-12-07

158

Mercury Binding Sites in Thiol-Functionalized Mesostructured Silica  

International Nuclear Information System (INIS)

Thiol-functionalized mesostructured silica with anhydrous compositions of (SiO2)1-x(LSiO1.5)x, where L is a mercaptopropyl group and x is the fraction of functionalized framework silicon centers, are effective trapping agents for the removal of mercuric(II) ions from water. In the present work, we investigate the mercury-binding mechanism for representative thiol-functionalized mesostructures by atomic pair distribution function (PDF) analysis of synchrotron X-ray powder diffraction data and by Raman spectroscopy. The mesostructures with wormhole framework structures and compositions corresponding to x = 0.30 and 0.50 were prepared by direct assembly methods in the presence of a structure-directing amine porogen. PDF analyses of five mercury-loaded compositions with Hg/S ratios of 0.50-1.30 provided evidence for the bridging of thiolate sulfur atoms to two metal ion centers and the formation of chain structures on the pore surfaces. We find no evidence for Hg-O bonds and can rule out oxygen coordination of the mercury at greater than the 10% level. The relative intensities of the PDF peaks corresponding to Hg-S and Hg-Hg atomic pairs indicate that the mercury centers cluster on the functionalized surfaces by virtue of thiolate bridging, regardless of the overall mercury loading. However, the Raman results indicate that the complexation of mercury centers by thiolate depends on the mercury loading. At low mercury loadings (Hg/S (le) 0.5), the dominant species is an electrically neutral complex in which mercury most likely is tetrahedrally coordinated to bridging thiolate ligands, as in Hg(SBut)2. At higher loadings (Hg/S 1.0-1.3), mercury complex cations predominate, as evidenced by the presence of charge-balancing anions (nitrate) on the surface. This cationic form of bound mercury is assigned a linear coordination to two bridging thiolate ligands.

2005-06-15

159

Advanced electron paramagnetic resonance on the catalytic iron-sulfur cluster bound to the CCG domain of heterodisulfide reductase and succinate: quinone reductase.  

UK PubMed Central (United Kingdom)

Heterodisulfide reductase (Hdr) is a key enzyme in the energy metabolism of methanogenic archaea. The enzyme catalyzes the reversible reduction of the heterodisulfide (CoM-S-S-CoB) to the thiol coenzymes M (CoM-SH) and B (CoB-SH). Cleavage of CoM-S-S-CoB at an unusual FeS cluster reveals unique substrate chemistry. The cluster is fixed by cysteines of two cysteine-rich CCG domain sequence motifs (CX31-39CCX35-36CXXC) of subunit HdrB of the Methanothermobacter marburgensis HdrABC complex. We report on Q-band (34 GHz) (57)Fe electron-nuclear double resonance (ENDOR) spectroscopic measurements on the oxidized form of the cluster found in HdrABC and in two other CCG-domain-containing proteins, recombinant HdrB of Hdr from M. marburgensis and recombinant SdhE of succinate: quinone reductase from Sulfolobus solfataricus P2. The spectra at 34 GHz show clearly improved resolution arising from the absence of proton resonances and polarization effects. Systematic spectral simulations of 34 GHz data combined with previous 9 GHz data allowed the unambiguous assignment of four (57)Fe hyperfine couplings to the cluster in all three proteins. (13)C Mims ENDOR spectra of labelled CoM-SH were consistent with the attachment of the substrate to the cluster in HdrABC, whereas in the other two proteins no substrate is present. (57)Fe resonances in all three systems revealed unusually large (57)Fe ENDOR hyperfine splitting as compared to known systems. The results infer that the cluster's unique magnetic properties arise from the CCG binding motif.

Fielding AJ; Parey K; Ermler U; Scheller S; Jaun B; Bennati M

2013-09-01

160

Identification of novel aroma-active thiols in pan-roasted white sesame seeds.  

Science.gov (United States)

Screening for aroma-active compounds in an aroma distillate obtained from freshly pan-roasted sesame seeds by aroma extract dilution analysis revealed 32 odorants in the FD factor range of 2-2048, 29 of which could be identified. The highest FD factors were found for the coffee-like smelling 2-furfurylthiol, the caramel-like smelling 4-hydroxy-2,5-dimethyl-3(2H)-furanone, the coffee-like smelling 2-thenylthiol (thiophen-2-yl-methylthiol), and the clove-like smelling 2-methoxy-4-vinylphenol. In addition, 9 odor-active thiols with sulfurous, meaty, and/or catty, black-currant-like odors were identified for the first time in roasted sesame seeds. Among them, 2-methyl-1-propene-1-thiol, (Z)-3-methyl-1-butene-1-thiol, (E)-3-methyl-1-butene-1-thiol, (Z)-2-methyl-1-butene-1-thiol, (E)-2-methyl-1-butene-1-thiol, and 4-mercapto-3-hexanone were previously unknown as food constituents. Their structures were confirmed by comparing their mass spectra and retention indices as well as their sensory properties with those of synthesized reference compounds. The relatively unstable 1-alkene-1-thiols represent a new class of food odorants and are suggested as the key contributors to the characteristic, but quickly vanishing, aroma of freshly ground roasted sesame seeds. PMID:20491509

Tamura, Hitoshi; Fujita, Akira; Steinhaus, Martin; Takahisa, Eisuke; Watanabe, Hiroyuki; Schieberle, Peter

2010-06-23

 
 
 
 
161

Identification of novel aroma-active thiols in pan-roasted white sesame seeds.  

UK PubMed Central (United Kingdom)

Screening for aroma-active compounds in an aroma distillate obtained from freshly pan-roasted sesame seeds by aroma extract dilution analysis revealed 32 odorants in the FD factor range of 2-2048, 29 of which could be identified. The highest FD factors were found for the coffee-like smelling 2-furfurylthiol, the caramel-like smelling 4-hydroxy-2,5-dimethyl-3(2H)-furanone, the coffee-like smelling 2-thenylthiol (thiophen-2-yl-methylthiol), and the clove-like smelling 2-methoxy-4-vinylphenol. In addition, 9 odor-active thiols with sulfurous, meaty, and/or catty, black-currant-like odors were identified for the first time in roasted sesame seeds. Among them, 2-methyl-1-propene-1-thiol, (Z)-3-methyl-1-butene-1-thiol, (E)-3-methyl-1-butene-1-thiol, (Z)-2-methyl-1-butene-1-thiol, (E)-2-methyl-1-butene-1-thiol, and 4-mercapto-3-hexanone were previously unknown as food constituents. Their structures were confirmed by comparing their mass spectra and retention indices as well as their sensory properties with those of synthesized reference compounds. The relatively unstable 1-alkene-1-thiols represent a new class of food odorants and are suggested as the key contributors to the characteristic, but quickly vanishing, aroma of freshly ground roasted sesame seeds.

Tamura H; Fujita A; Steinhaus M; Takahisa E; Watanabe H; Schieberle P

2010-06-01

162

Species variations in biliary excretion of glutathione-related thiols and methylmercury.  

Science.gov (United States)

The biliary excretion of methylmercury is thought to be related to the biliary excretion of nonprotein thiols in rats. Species differences in biliary excretion of glutathione (GSH) and related thiols are unknown; therefore, the relationship between the biliary excretion of GSH-related thiols and methylmercury in five species was studied. The biliary excretion rate of GSH-related thiols and disulfides was 369, 192, 94, 50, and 19 nmol/min/kg for mice, rats, hamsters, guinea pigs, and rabbits, respectively. The main thiol in mouse, hamster, and rat bile was GSH, whereas guinea pig and rabbit bile contained mainly cysteinylglycine (Cys-Gly). The larger percentage of Cys-Gly in guinea pig and rabbit bile was correlated with their greater hepatic gamma-glutamyltranspeptidase (GGT) activity than that observed in the other species. The biliary excretion rate (nmol/min/kg) of methylmercury was approximately 0.8 in mice, rats, and hamsters compared to significantly lower rates in guinea pigs and rabbits (0.15 and 0.03, respectively). It is concluded that the species-specific composition of GSH-related thiols and disulfides in bile is related to species variations in hepatic GGT activity, and that the species variation in biliary excretion of GSH-related thiols does not entirely account for the species variation in methylmercury excretion, indicating other factors are also apparently involved in determining the rate of biliary excretion of methylmercury. PMID:2897139

Stein, A F; Gregus, Z; Klaassen, C D

1988-05-01

163

Conductive probe AFM study of Pt-thiol and Au-thiol contacts in metal-molecule-metal systems.  

UK PubMed Central (United Kingdom)

The charge transport mechanism between 1,8-octanedithiol (ODT, C(8)H(16)S(2)H(2)) and platinum and gold electrodes is studied by breaking bonds between single ODT molecules and atomic metal junctions using conductive probe atomic force microscopy. Histograms of conductance values show peaks that are obscured by background processes that differ from the metal-molecule-metal conduction path of interest. We introduce a new method to reduce greatly such backgrounds by dividing by a 1-octanethiol (OMT, C(8)H(17)SH) reference histogram, without data selection. The method reveals three series of conductance values for both platinum and gold contacts, which we associate with geometrically different configurations between thiol and metal atoms. The ordering of conductance values, Pt-ODT-Pt > Pt-ODT-Au> Au-ODT-Au, is consistent with a relative dependence on both the number of electron channels and the density of states.

Kim CM; Bechhoefer J

2013-01-01

164

Respiratory arsenate reductase as a bidirectional enzyme  

Science.gov (United States)

The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe–S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism.

Richey, C.; Chovanec, P.; Hoeft, S. E.; Oremland, R. S.; Basu, P.; Stolz, J. F.

2009-01-01

165

Respiratory arsenate reductase as a bidirectional enzyme.  

Science.gov (United States)

The haloalkaliphilic bacterium Alkalilimnicola ehrlichii is capable of anaerobic chemolithoautotrophic growth by coupling the oxidation of arsenite (As(III)) to the reduction of nitrate and carbon dioxide. Analysis of its complete genome indicates that it lacks a conventional arsenite oxidase (Aox), but instead possesses two operons that each encode a putative respiratory arsenate reductase (Arr). Here we show that one homolog is expressed under chemolithoautotrophic conditions and exhibits both arsenite oxidase and arsenate reductase activity. We also demonstrate that Arr from two arsenate respiring bacteria, Alkaliphilus oremlandii and Shewanella sp. strain ANA-3, is also biochemically reversible. Thus Arr can function as a reductase or oxidase. Its physiological role in a specific organism, however, may depend on the electron potentials of the molybdenum center and [Fe-S] clusters, additional subunits, or constitution of the electron transfer chain. This versatility further underscores the ubiquity and antiquity of microbial arsenic metabolism. PMID:19285953

Richey, Christine; Chovanec, Peter; Hoeft, Shelley E; Oremland, Ronald S; Basu, Partha; Stolz, John F

2009-03-13

166

Primary structure of chicken liver dihydrofolate reductase.  

UK PubMed Central (United Kingdom)

The complete covalent structure of dihydrofolate reductase from chicken liver is described. The S-carboxymethylated protein was subjected to cleavage by cyanogen bromide which produced five fragments. Fragment CB2 contained an internal homoserine residue which was not cleaved by cyanogen bromide. Sequences and ordering of the cyanogen bromide fragments were established by means of automated sequencer analyses of the fragments and from smaller peptides generated by proteolysis with trypsin and staphylococcal protease. The covalent structure of the single polypeptide chain comprises 189 residues of molecular weight 21,651. The chicken liver enzyme is homologous to that from L1210 cells and shows regions of homology to dihydrofolate reductases from Streptococcus faecium, Escherichia coli, and Lactobacillus casei. These homologous regions in the chicken liver enzyme are primarily related to conserved amino acid residues implicated in the binding of NADPH and methotrexate by bacterial dihydrofolate reductases.

Kumar AA; Blankenship DT; Kaufman BT; Freisheim JH

1980-02-01

167

Evaluation of nitrate reductase activity in Rhizobium japonicum  

Energy Technology Data Exchange (ETDEWEB)

Nitrate reductase activity was evaluated by four approaches, using four strains of Rhizobium japonicum and 11 chlorate-resistant mutants of the four strains. It was concluded that in vitro assays with bacteria or bacteroids provide the most simple and reliable assessment of the presence or absence of nitrate reductase. Nitrite reductase activity with methyl viologen and dithionite was found, but the enzyme activity does not confound the assay of nitrate reductase. 18 references

Streeter, J.G.; DeVine, P.J.

1983-08-01

168

Nitrate reductase of green algae is located in the pyrenoid.  

UK PubMed Central (United Kingdom)

Antibodies against nitrate reductase from Monoraphidium braunii have been used to determine the antigenic relationships of nitrate reductases from different green algae. Nitrate reductases from Chlamydomonas reinhardii, Chlorella fusca, Dunaliella salina, and Scenedesmus obliquus, were inhibited by, and cross-reacted with, antibodies raised against the enzyme from Monoraphidium braunii.These antibodies were also used to determine, by immunoelectron microscopy, the intracellular location of nitrate reductase in the aforementioned green algae. In all cases, the enzyme was specifically located in the pyrenoid.

Lopez-Ruiz A; Verbelen JP; Roldan JM; Diez J

1985-12-01

169

Metmyoglobin reductase activity in bovine muscles.  

UK PubMed Central (United Kingdom)

Cow muscles (Longissimus dorsi, Tensor fasciae latae, Psoas major, Diaphragma medialis), with different colour stabilities, were used to measure 'Metmyoglobin reductase activity' in different conditions. The effects of pH and temperature on in vitro metmyoglobin reduction were analysed. The highest metmyoglobin reductase activity was localized in microsomes and in more or less intact mitochondria by means of differential centrifugations. The most unstable muscles, from the colour point of view, presented the highest reducing activities and no differences were noted between activities measured in aerobic or anaerobic conditions.

Echevarne C; Renerre M; Labas R

1990-01-01

170

Ebsulfur is a benzisothiazolone cytocidal inhibitor targeting the trypanothione reductase of Trypanosoma brucei.  

UK PubMed Central (United Kingdom)

Trypanosoma brucei is the etiological agent of African trypanosomiasis. These parasites possess a unique thiol redox system that includes trypanothione (T(SH)2) and trypanothione reductase (TryR) instead of the thioredoxin and glutaredoxin systems of mammalian hosts required for DNA synthesis and defense against oxidative stress. Here we show that benzisothiazolone ebsulfur (EbS), a sulfur analogue of ebselen, is a potent inhibitor of T. brucei growth with favourable selective index over mammalian cells. EbS inhibited the TryR activity and decreased non-protein thiol levels in cultured parasites. The inhibition of TryR by EbS was irreversible, and NADPH-dependent. EbS formed a complex with TryR and caused oxidation and inactivation of the enzyme. EbS was more toxic for T. brucei than for T. cruzi, probably due to lower levels of TryR and T(SH)2 in T. brucei. Furthermore, inhibition of TryR produced high intracellular ROS. Hydrogen peroxide, known to be constitutively high in T. brucei, enhanced the EbS inhibition of TryR. The elevation of ROS production in parasites caused by EbS induced a programmed cell death. Soluble EbS analogues were synthesized and cured T. b. brucei-infection in mice when inoculated with nifurtimox. Altogether, EbS and EbS analogues disrupt the trypanothione system impeding the defense against oxidative stress. Thus EbS is a promising lead for development of drugs against African trypanosomiasis.

Lu J; Vodnala SK; Gustavsson AL; Gustafsson TN; Sjoberg B; Johansson HA; Kumar S; Tjernberg A; Engman L; Rottenberg ME; Holmgren A

2013-07-01

171

Ebsulfur Is a Benzisothiazolone Cytocidal Inhibitor Targeting the Trypanothione Reductase of Trypanosoma brucei.  

UK PubMed Central (United Kingdom)

Trypanosoma brucei is the causing agent of African trypanosomiasis. These parasites possess a unique thiol redox system required for DNA synthesis and defense against oxidative stress. It includes trypanothione and trypanothione reductase (TryR) instead of the thioredoxin and glutaredoxin systems of mammalian hosts. Here, we show that the benzisothiazolone compound ebsulfur (EbS), a sulfur analogue of ebselen, is a potent inhibitor of T. brucei growth with a favorable selectivity index over mammalian cells. EbS inhibited the TryR activity and decreased non-protein thiol levels in cultured parasites. The inhibition of TryR by EbS was irreversible and NADPH-dependent. EbS formed a complex with TryR and caused oxidation and inactivation of the enzyme. EbS was more toxic for T. brucei than for Trypanosoma cruzi, probably due to lower levels of TryR and trypanothione in T. brucei. Furthermore, inhibition of TryR produced high intracellular reactive oxygen species. Hydrogen peroxide, known to be constitutively high in T. brucei, enhanced the EbS inhibition of TryR. The elevation of reactive oxygen species production in parasites caused by EbS induced a programmed cell death. Soluble EbS analogues were synthesized and cured T. brucei brucei infection in mice when used together with nifurtimox. Altogether, EbS and EbS analogues disrupt the trypanothione system, hampering the defense against oxidative stress. Thus, EbS is a promising lead for development of drugs against African trypanosomiasis.

Lu J; Vodnala SK; Gustavsson AL; Gustafsson TN; Sjöberg B; Johansson HA; Kumar S; Tjernberg A; Engman L; Rottenberg ME; Holmgren A

2013-09-01

172

Ebsulfur Is a Benzisothiazolone Cytocidal Inhibitor Targeting the Trypanothione Reductase of Trypanosoma brucei.  

Science.gov (United States)

Trypanosoma brucei is the causing agent of African trypanosomiasis. These parasites possess a unique thiol redox system required for DNA synthesis and defense against oxidative stress. It includes trypanothione and trypanothione reductase (TryR) instead of the thioredoxin and glutaredoxin systems of mammalian hosts. Here, we show that the benzisothiazolone compound ebsulfur (EbS), a sulfur analogue of ebselen, is a potent inhibitor of T. brucei growth with a favorable selectivity index over mammalian cells. EbS inhibited the TryR activity and decreased non-protein thiol levels in cultured parasites. The inhibition of TryR by EbS was irreversible and NADPH-dependent. EbS formed a complex with TryR and caused oxidation and inactivation of the enzyme. EbS was more toxic for T. brucei than for Trypanosoma cruzi, probably due to lower levels of TryR and trypanothione in T. brucei. Furthermore, inhibition of TryR produced high intracellular reactive oxygen species. Hydrogen peroxide, known to be constitutively high in T. brucei, enhanced the EbS inhibition of TryR. The elevation of reactive oxygen species production in parasites caused by EbS induced a programmed cell death. Soluble EbS analogues were synthesized and cured T. brucei brucei infection in mice when used together with nifurtimox. Altogether, EbS and EbS analogues disrupt the trypanothione system, hampering the defense against oxidative stress. Thus, EbS is a promising lead for development of drugs against African trypanosomiasis. PMID:23900839

Lu, Jun; Vodnala, Suman K; Gustavsson, Anna-Lena; Gustafsson, Tomas N; Sjöberg, Birger; Johansson, Henrik A; Kumar, Sangit; Tjernberg, Agneta; Engman, Lars; Rottenberg, Martin E; Holmgren, Arne

2013-07-29

173

Enhancing effect of a cysteinyl thiol on the antioxidant activity of flavonoids and identification of the antioxidative thiol adducts of myricetin.  

UK PubMed Central (United Kingdom)

The enhancing effect of a cysteinyl thiol N-benzoylcysteine methyl ester on the antioxidant activity of several flavonoids was investigated in a lipid oxidation system. Obvious enhancement was apparent for catechin, myricetin, quercetin, and taxifolin, the activity for myricetin being the most potent among them. An HPLC analysis of the products from the antioxidation reaction of myricetin in the presence of the thiol was carried out and the structures of the products were determined to clarify the enhancing effect chemically. The obtained data indicated that two thiol adducts on the B ring, and probably C-ring adducts, which were produced in the antioxidation process, exerted an enhancing effect on the antioxidant activity of myricetin.

Masuda T; Miura Y; Inai M; Masuda A

2013-08-01

174

Thiol-Ene Based Polymer Waveguides Fabricated By Uv-Assisted Soft Lithography For Optofluidic Applications  

DEFF Research Database (Denmark)

In this paper, a thiol-ene based polymer waveguide, defined by UV-assisted soft lithography, is designed, fabricated and characterized. Waveguides are formed by filling microfluidic channels with a high refractive index liquid mixture of ‘thiol’ and ‘ene’ monomers (e.g., trimethylolpropane tris(3-mercaptopropionate) = ‘thiol’, and 1,3,5-triallyl-1,3,5-triazine-2,4,6(1H,3H,5H)-trione = ‘ene’), which can be cured by UV exposure into a solid polymer. The waveguides demonstrated good confinement of light, and a propagation loss of 0.5 dB/cm was obtained. To our best knowledge, this is the first report to employ thiol-ene based polymers as waveguide core materials for potential optofluidic applications.

Zhuang, Guisheng; Jensen, Thomas Glasdam

2011-01-01

175

AFM-assisted fabrication of thiol SAM pattern with alternating quantified surface potential  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Thiol self-assembled monolayers (SAMs) are widely used in many nano- and bio-technology applications. We report a new approach to create and characterize a thiol SAMs micropattern with alternating charges on a flat gold-coated substrate using atomic force microscopy (AFM) and Kelvin probe force microscopy (KPFM). We produced SAMs-patterns made of alternating positively charged, negatively charged, and hydrophobic-terminated thiols by an automated AFM-assisted manipulation, or nanografting. We show that these thiol patterns possess only small topographical differences as revealed by AFM, and distinguished differences in surface potential (20-50 mV), revealed by KPFM. The pattern can be helpful in the development of biosensor technologies, specifically for selective binding of biomolecules based on charge and hydrophobicity, and serve as a model for creating surfaces with quantified alternating surface potential distribution.

Moores Bradley; Simons Janet; Xu Song; Leonenko Zoya

2011-01-01

176

Monolayers of thiol-terminated dendrimers on the surface of planar and colloidal gold  

Energy Technology Data Exchange (ETDEWEB)

The synthesis of fourth-generation poly(amido-amine)(PAMAM) dendrimers having terminal groups partially or fully functionalized with thiol groups is described. These thiolated dendrimers form stable monolayers on planar Au substrates. X-ray photoelectron spectroscopy studies reveal that most of the thiol groups on dendrimers having 20% of their terminal primary amine groups functionalized can directly interact with the Au furnace. This suggests that the dendrimer molecules are flexible and arrange themselves so as to maximize the number of Au/thiol interactions. Reduction of tetrachloroauric acid in the presence of thiol-modified dendrimers results in formation of water-soluble, dendrimer-stabilized nanoparticles. The particles are 1.5--2.1 nm in diameter depending on the Au/dendrimer ratio used. Stable nanoparticles can be obtained at Au/dendrimer ratios as large as 120:1.

Chechik, V.; Crooks, R.M.

1999-09-14

177

New functional materials for heavy metal sorption: "Supramolecular" attachment of thiols to mesoporous silica substrates  

Science.gov (United States)

A new class of sorbent material, which exhibits exceptional metal capture from contaminated natural water, features aromatic thiol ligands reversibly bound to functionalized mesoporous silica through non-covalent interactions and have the potential of being regenerable.

Carter, Timothy G.; Yantasee, Wassana; Sangvanich, Thanapon; Fryxell, Glen E.; Johnson, Darren W.; Addleman, R. Shane

2008-01-01

178

Influence of thiols and oxygen on the survival of ?-irradiated plasmid DNA and cells  

Science.gov (United States)

Some of the recent progress made in the understanding of the quantitative aspects of the oxygen effect in radiation biology by several groups is summarized. Examples are: the importance of unrepairable damage for the quantitative description of the oxygen effect; proof that protein thiols hardly contribute to protection in cells in the absence of oxygen; the proposal that protection by thiols in concentration ranges where al DNA radicals react with oxygen is due to the formation of hydroperoxides which can be repaired enzymatically by glutathione peroxydase; the finding that unscavengeable damage in plasmid DNA is mainly due to spur-induced clustered damages, but that the precursors of the scavengeable and the unscavengeable damage are comparably well repaired by thiols; the result that E. coli repair wild type strains are better protected by addition of thiols than strains with deficiencies in enzymatic repair capacities.

Schulte-Frohlinde, D.; Ludwig, D. C.; Rettberg, P.

1994-10-01

179

Influence of thiols and oxygen on the survival of gamma-irradiated plasmid DNA and cells.  

Science.gov (United States)

Some of the recent progress made in the understanding of the quantitative aspects of the oxygen effect in radiation biology by several groups is summarized. Examples are: the importance of unrepairable damage for the quantitative description of the oxygen effect; proof that protein thiols hardly contribute to protection in cells in the absence of oxygen; the proposal that protection by thiols in concentration ranges where all DNA radicals react with oxygen is due to the formation of hydroperoxides which can be repaired enzymatically by glutathione peroxydase; the finding that unscavengeable damage in plasmid DNA is mainly due to spur-induced clustered damages, but that the precursors of the scavengeable and the unscavengeable damage are comparably well repaired by thiols; the result that E. coli repair wild type strains are better protected by addition of thiols than strains with deficiencies in enzymatic repair capacities. PMID:11539962

Schulte-Frohlinde, D; Ludwig, D C; Rettberg, P

1994-10-01

180

Studies of thiol additions of silane coupling agents by vibrational spectroscopy.  

UK PubMed Central (United Kingdom)

Raman and infrared spectroscopies have been used to determine the addition reaction of mercaptopropyltrimethoxysilane (MPTMS) to allyltrimethoxysilane (ATMS) and 7-octenyltrichlorosilane based on the vibrational intensity variation of thiol and vinyl groups in the reaction mixtures. Due to the distinct and moderate intensity of Raman bands observed in the present experiment, the identification with Raman spectroscopic method is more sensitive than that with FTIR spectroscopy. In the presence of UV radiation, thiol addition reaction has been observed in the direct mixing samples of silanes. Hybrid sol-gels prepared with the use of MPTMS and ATMS as precursors in both acidic and basic conditions have revealed the progression of thiol addition under the UV radiation exposure. UV radiation is similarly effective to induce the thiol addition in the sol-gel coated aluminum tiles. Without UV radiation, the use of free radical initiator in the sol-gel samples might also help to induce the addition reaction.

Li YS; Vecchio NE

2007-08-01

 
 
 
 
181

Maleimide-functionalised organoruthenium anticancer agents and their binding to thiol-containing biomolecules.  

UK PubMed Central (United Kingdom)

Ru(II)(arene) anticancer compounds with maleimide functionality were prepared to allow selective interaction with thiol-containing biomolecules and thereby enforcing the selective delivery of the compounds to the tumour.

Hanif M; Nazarov AA; Legin A; Groessl M; Arion VB; Jakupec MA; Tsybin YO; Dyson PJ; Keppler BK; Hartinger CG

2012-02-01

182

Content of endogenous thiols and radioresistance of gemmating cells of Saccharomyces ellipsoideus and Saccharomyces cerevisiale yeasts  

International Nuclear Information System (INIS)

It has been shown that gemmating cells of ''wild type'' yeasts are more radioresistant and contain more endogenous thiols, than resting cells. Gemmating cells of Saccharomyces cerevisial yeasts, carrying the mutation rad 51, as to radioresistance and content of SH groups do not differ from resting cells. The results obtained testify to a connec-- tion between increased radioresistance of the yeast gemmating cells and increased content of endogenous thiols in them

1983-01-01

183

Testing of thiol-terminated oligonucleotides as potential DNA-cleaving reagents  

Energy Technology Data Exchange (ETDEWEB)

In this study the authors have conducted experiments to determine if oligonucleotides containing terminal thiol groups can be used, in the presence of Cu+2, to cut single- and double-stranded DNA molecules adjacent to the oligonucleotide binding site. The results of these experiments demonstrate that these modified oligonucleotides cleave DNA in the presence of Cu+2, but the reaction could not be controlled sufficiently to attain site-selective cleavage or minimize degradation of the thiol terminated oligonucleotide.

Balhorn, R.; Mazrimas, J. [Lawrence Livermore National Lab., CA (United States). Biology and Biotechnology Research Program

1996-06-01

184

Evolving role of ribonucleoside reductase inhibitors in hematologic malignancies.  

UK PubMed Central (United Kingdom)

Ribonucleotide reductases catalyze the de novo biosynthesis of deoxyribonucleosides for DNA synthesis. Increased ribonucleotide reductases activity has been associated with malignant transformation and tumor cell growth. The ribonucleotide reductases inhibitors may bind with the R1 subunit of the enzyme (Class 1) or the nonheme iron (Class 2). This review focuses on the therapeutic use of ribonucleotide reductases inhibitors in hematologic malignancies. Hydroxyurea, fludarabine and cladribine have established roles in the management of hematologic malignancies, while other ribonucleotide reductases inhibitors, such as gemcitabine, tezacitabine and heterocyclic carboxaldehyde thiosemicarbazones (e.g., triapine) are being evaluated in clinical trials.

Tsimberidou AM; Alvarado Y; Giles FJ

2002-08-01

185

Evolving role of ribonucleoside reductase inhibitors in hematologic malignancies.  

Science.gov (United States)

Ribonucleotide reductases catalyze the de novo biosynthesis of deoxyribonucleosides for DNA synthesis. Increased ribonucleotide reductases activity has been associated with malignant transformation and tumor cell growth. The ribonucleotide reductases inhibitors may bind with the R1 subunit of the enzyme (Class 1) or the nonheme iron (Class 2). This review focuses on the therapeutic use of ribonucleotide reductases inhibitors in hematologic malignancies. Hydroxyurea, fludarabine and cladribine have established roles in the management of hematologic malignancies, while other ribonucleotide reductases inhibitors, such as gemcitabine, tezacitabine and heterocyclic carboxaldehyde thiosemicarbazones (e.g., triapine) are being evaluated in clinical trials. PMID:12647987

Tsimberidou, Apostolia-Maria; Alvarado, Yesid; Giles, Francis J

2002-08-01

186

Facile functionalization of PDMS elastomer surfaces using thiol-ene click chemistry.  

UK PubMed Central (United Kingdom)

A variety of methods have been developed for polydimethylsiloxane (PDMS) elastomer surface functionalization, particularly for the improvement of hydrophilicity. However, in addition to difficulties in avoiding undesired physical changes to the modified surface, including surface cracking, "hydrophobic recovery" frequently leads hydrophilically modified surfaces to completely return over time to their hydrophobic nature, with accompanying loss of accessible functional groups. Thiol-ene chemistry provides a mild and robust technology for synthetic elaboration. We demonstrate the introduction of thiol groups onto the PDMS surface via base-catalyzed equilibration of MTS ((MeO)3Si(CH2)3SH). Thiols in the product elastomer were shown to be located primarily at the air interface using EDX, XPS and fluorescence labeling initially, and after extended periods of time: total thiol concentrations at the surface and in the bulk were established by complementary chemical titrations with DTDP (4,4'-dithiodipyridine) and iodine titrations in different solvents. The surface density of thiols was readily controlled by reaction conditions: the rate of hydrophobic recovery, which led to incomplete loss of accessible functional groups, was determined. Thiol-ene click chemistry was then used to introduce a variety of hydrophilic moieties onto the surface including a silicone surfactant and maleic anhydride, respectively. In the latter case, molecular functionalization with both small (fluorescent labels) and polymeric nucleophiles (poly(ethylene glycol), chitosan) could be subsequently induced by simple ring-opening nucleophilic attack leading to permanently functional surfaces.

Zhang J; Chen Y; Brook MA

2013-09-01

187

Integration of the thiol redox status with cytokine response to physical training in professional basketball players.  

Science.gov (United States)

The present study was designed to evaluate the plasma markers of reactive oxygen species (ROS) activity and cytokines, and their relationship with thiol redox status of basketball players during training. Sixteen professional players of the Polish Basketball Extraleague participated in the study. The study was performed during the preparatory period and the play-off round. Markers of ROS activity (lipid peroxidation TBARS, protein carbonylation PC) and reduced glutathione (GSH) demonstrated regularity over time, i.e. TBARS, PC and GSH were elevated at the beginning and decreased at the end of training periods. Oxidized glutathione (GSSG) was not affected by exercise training. Thiol redox status (GSH(total)-2GSSG/GSSG) correlated with TBARS and PC in both training periods. The level of interleukin-6 (IL-6) was increased and positively correlated with thiol redox (r=0.423) in the preparatory period, whereas tumor necrosis factor alpha (TNFalpha) was increased and inversely correlated with thiol redox (r= 0.509) in the play-off round. The present study showed significant shifts in markers of ROS activity, thiol redox status and inflammatory mediators (IL-6, TNFalpha) following professional sport training as well as correlation between changes in thiol redox and cytokine response. PMID:19537921

Zembron-Lacny, A; Slowinska-Lisowska, M; Ziemba, A

2009-06-19

188

Integration of the thiol redox status with cytokine response to physical training in professional basketball players.  

UK PubMed Central (United Kingdom)

The present study was designed to evaluate the plasma markers of reactive oxygen species (ROS) activity and cytokines, and their relationship with thiol redox status of basketball players during training. Sixteen professional players of the Polish Basketball Extraleague participated in the study. The study was performed during the preparatory period and the play-off round. Markers of ROS activity (lipid peroxidation TBARS, protein carbonylation PC) and reduced glutathione (GSH) demonstrated regularity over time, i.e. TBARS, PC and GSH were elevated at the beginning and decreased at the end of training periods. Oxidized glutathione (GSSG) was not affected by exercise training. Thiol redox status (GSH(total)-2GSSG/GSSG) correlated with TBARS and PC in both training periods. The level of interleukin-6 (IL-6) was increased and positively correlated with thiol redox (r=0.423) in the preparatory period, whereas tumor necrosis factor alpha (TNFalpha) was increased and inversely correlated with thiol redox (r= 0.509) in the play-off round. The present study showed significant shifts in markers of ROS activity, thiol redox status and inflammatory mediators (IL-6, TNFalpha) following professional sport training as well as correlation between changes in thiol redox and cytokine response.

Zembron-Lacny A; Slowinska-Lisowska M; Ziemba A

2010-01-01

189

Thiols as mechanistic probes for catalysis by the free radical enzyme galactose oxidase.  

UK PubMed Central (United Kingdom)

Galactose oxidase, a mononuclear copper enzyme, oxidizes primary alcohols to aldehydes using molecular oxygen. A unique type of cross-link between tyrosine 272, an active-site copper ligand, and cysteine 228 provides a modified tyrosine radical site believed to act as a one-electron redox center. Substrate analogs incorporating a primary thiol group in place of the primary alcohol group in normal substrates (RCH2OH) have been studied as active-site mechanistic probes. Thiol sulfur coordinates to the active-site copper, leading to enzyme inactivation in a time- and concentration-dependent manner. The mechanism of inactivation involves redox chemistry related to the active-site redox centers, though inactivation does not proceed through the rate-determining hydrogen atom abstraction step that occurs in alcohol oxidation. Thiols are therefore classified as active-site-directed redox inactivators. The thiol analog of galactose, 6-Thio-Me-Gal, is also turned over by the enzyme, albeit at a much reduced rate, indicating that the energetics of turnover is changed significantly. Thiols constitute a particularly good model of the ground state enzyme-substrate complex. The Michaelis complex for thiol substrate analogs is stabilized at least 200-fold compared to the analogous alcohol substrates, whereas the transition state of H atom abstraction is destabilized, presumably due to a slight increase in distances of reacting atoms and weakening of hydrogen-bonding interactions due to the larger atomic radius of sulfur compared to that of oxygen.

Wachter RM; Branchaud BP

1996-11-01

190

Thiols as mechanistic probes for catalysis by the free radical enzyme galactose oxidase.  

Science.gov (United States)

Galactose oxidase, a mononuclear copper enzyme, oxidizes primary alcohols to aldehydes using molecular oxygen. A unique type of cross-link between tyrosine 272, an active-site copper ligand, and cysteine 228 provides a modified tyrosine radical site believed to act as a one-electron redox center. Substrate analogs incorporating a primary thiol group in place of the primary alcohol group in normal substrates (RCH2OH) have been studied as active-site mechanistic probes. Thiol sulfur coordinates to the active-site copper, leading to enzyme inactivation in a time- and concentration-dependent manner. The mechanism of inactivation involves redox chemistry related to the active-site redox centers, though inactivation does not proceed through the rate-determining hydrogen atom abstraction step that occurs in alcohol oxidation. Thiols are therefore classified as active-site-directed redox inactivators. The thiol analog of galactose, 6-Thio-Me-Gal, is also turned over by the enzyme, albeit at a much reduced rate, indicating that the energetics of turnover is changed significantly. Thiols constitute a particularly good model of the ground state enzyme-substrate complex. The Michaelis complex for thiol substrate analogs is stabilized at least 200-fold compared to the analogous alcohol substrates, whereas the transition state of H atom abstraction is destabilized, presumably due to a slight increase in distances of reacting atoms and weakening of hydrogen-bonding interactions due to the larger atomic radius of sulfur compared to that of oxygen. PMID:8916929

Wachter, R M; Branchaud, B P

1996-11-12

191

The role of the thiol group in protein modification with methylglyoxal  

Directory of Open Access Journals (Sweden)

Full Text Available Methylglyoxal is a highly reactive ?-oxoaldehyde with elevated production in hyperglycemia. It reacts with nucleophilic Lys and Arg side-chains and N-terminal amino groups causing protein modification. In the present study, the importance of the reaction of the Cys thiol group with methylglyoxal in protein modification, the competitiveness of this reaction with those of amino and guanidine groups, the time course of these reactions and their role and contribution to protein cross-linking were investigated. Human and bovine serum albumins were used as model systems. It was found that despite the very low levels of thiol groups on the surface of the examined protein molecules (approx. 80 times lower than those of amino and guanidino groups), a very high percentage of it reacts (25–85 %). The amount of reacted thiol groups and the rate of the reaction, the time for the reaction to reach equilibrium, the formation of a stable product and the contribution of thiol groups to protein cross-linking depend on the methylglyoxal concentration. The product formed in the reaction of thiol and an insufficient quantity of methylglyoxal (compared to the concentrations of the groups accessible for modification) participates to a significant extent (4 %) to protein cross-linking. Metformin applied in equimolar concentration with methylglyoxal prevents its reaction with amino and guanidino groups but, however, not with thiol groups.

JELENA M. A?IMOVI?; BOJANA D. STANIMIROVI?; LJUBA M. MANDI?

2009-01-01

192

Thiol-disulfide exchange in signaling: disulfide bonds as a switch.  

UK PubMed Central (United Kingdom)

The major function of disulfide bonds is not only the stabilization of protein structures. Over the last 30 years, a change in perspective took place driven by groundbreaking experiments, which promoted disulfide bonds to central players in essential thiol-disulfide exchange reactions involved in signal transduction, thiol protection, and redox homeostasis regulation. This new view stimulated redox research and led to the discovery of novel redox pathways, redox enzymes, and new low-molecular-weight thiols. These redox-sensitive molecules operate along diverse pathways via a dynamic thiol-disulfide mechanism in which disulfide bonds are reversibly formed and reduced, thereby switching the molecules between different conformational and functional states. It is now clear that disulfide bonds play a pivotal role in cellular reduction and oxidation processes. However, in spite of the fundamental cell biological and medical importance of the thiol-disulfide exchange switches, we are only beginning to understand their principles of specificity, their mechanism of action, and their role in signal transduction. Our further progress in understanding the thiol-disulfide switches will strongly depend on the chemical tools and on the technological advances that will be made in the development of new methodologies.

Messens J; Collet JF

2013-05-01

193

Submerged nanocontact printing (SnCP) of thiols.  

UK PubMed Central (United Kingdom)

Biological patterned surfaces having sub-micron scale resolution are of great importance in many fields of life science and biomedicine. Different techniques have been proposed for surface patterning at the nanoscale. However, most of them present some limitations regarding the patterned area size or are time-consuming. Micro/nanocontact printing is the most representative soft lithography-based technique for surface patterning at the nanoscale. Unfortunately, conventional micro/nanocontact printing also suffers from problems such as diffusion and stamp collapsing that limit pattern resolution. To overcome these problems, a simple way of patterning thiols under liquid media using submerged nanocontact printing (SnCP) over large areas (approximately cm2) achieving nanosize resolution is presented. The technique is also low cost and any special equipment neither laboratory conditions are required. Nanostructured poly(dimethyl siloxane) stamps are replicated from commercially available digital video disks. SnCP is used to stamp patterns of 200 nm 1-octadecanethiol lines in liquid media, avoiding ink diffusion and stamp collapsing, over large areas on gold substrates compared with conventional procedures. Atomic force microscopy measurements reveal that the patterns have been successfully transferred with high fidelity. This is an easy, direct, effective and low cost methodology for molecule patterning immobilization which is of interest in those areas that require nanoscale structures over large areas, such as tissue engineering or biosensor applications.

Caballero D; Samitier J; Errachid A

2009-11-01

194

Tip-enhanced Raman spectroscopic imaging of patterned thiol monolayers  

Directory of Open Access Journals (Sweden)

Full Text Available Full spectroscopic imaging by means of tip-enhanced Raman spectroscopy (TERS) was used to measure the distribution of two isomeric thiols (2-mercaptopyridine (2-PySH) and 4-mercaptopyridine (4-PySH)) in a self-assembled monolayer (SAM) on a gold surface. From a patterned sample created by microcontact printing, an image with full spectral information in every pixel was acquired. The spectroscopic data is in good agreement with the expected molecular distribution on the sample surface due to the microcontact printing process. Using specific marker bands at 1000 cm?1 for 2-PySH and 1100 cm?1 for 4-PySH, both isomers could be localized on the surface and semi-quantitative information was deduced from the band intensities. Even though nanometer size resolution information was not required, the large signal enhancement of TERS was employed here to detect a monolayer coverage of weakly scattering analytes that were not detectable with normal Raman spectroscopy, emphasizing the usefulness of TERS.

Johannes Stadler; Thomas Schmid; Lothar Opilik; Phillip Kuhn; Petra S. Dittrich; Renato Zenobi

2011-01-01

195

Processing and targeting of the thiol protease aleurain: Progress report  

Energy Technology Data Exchange (ETDEWEB)

This study addresses the processing and targeting of the thiol protease aleurain in monocots. A probe derived from the aleurain cDNA specific for the 5'-most 400 bp (a region encoding the first 140 amino acids of the preprotein hybridized to at least 3 separate elements in the barley genome; only one represented the aleurain gene. In contrast, a probe specific for the remaining 2/23 of the cDNA (representing the protease domain) hybridized to only a single copy sequence. To know if this pattern pertained in other, closely related, monocots, we probed Southern blots of genomic DNA from maize, rye, oats, sorghum, and pearl millet with each probe. In each instance except for maize DNA, the 5' domain probe hybridizes to several fragments in addition to those identified by the protease domain probe. Presumable the darkest hybridization in each represents the fragment carrying the sequences homologous to barley aleurain. The fragments from a given restriction enzyme identified by the protease domain probe in sorghum, millet, and maize, were indistinguishable in size indicating that the gene sequences, as well as flanking DNA, are so well conserved among the group that the location of the hexanucleotide sequences have not diverged. (3 refs., 3 figs.)

Rogers, J.C.

1988-01-01

196

Human erythrocyte thiol methyltransferase: radiochemical microassay and biochemical properties  

International Nuclear Information System (INIS)

A radiochemical microassay for the measurement of thiol methyltransferase (TMT) activity in human red blood cell (RBC) membranes has been developed. Both 2-mercaptoethanol and dithiothreitol were used as substrates for the enzyme. The pH optimum of the reaction was approximately 9.0 when glycine-NaOH was used as a buffer. The apparent Michaelis-Menten (Ksub(M)) value for the methyl donor for the reaction, S-adenosyl-L-methionine, was 43 ?mol/l. Human RBC TMT activity was neither activated nor inhibited by Ca2+, Mg2+, or tropolone, but the enzyme was inhibited by SKF 525A and by reagents that react with sulfhydryl grcups. The mean TMT activity in blood from 289 randomly selected adult white subjects was 10.93 +- 3.22 units per mg protein (mean +- S.D.). The activity was the same in samples from men and women. The results of experiments in which TMT activity was measured in mixtures of RBC membranes with relatively ''low'' and relatively ''high'' activities provided no evidence that individual variations in the enzyme activity were due to variations in endogenous TMT activators or inhibitors. (Auth.)

1979-09-15

197

Oligomerization of Indole Derivatives with Incorporation of Thiols  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract: Two molecules of indole derivative, e.g. indole-5-carboxylic acid, reacted with one molecule of thiol, e.g. 1,2-ethanedithiol, in the presence of trifluoroacetic acid to yield adducts such as 3-[2-(2-amino-5-carboxyphenyl)-1-(2-mercaptoethylthio)ethyl]-1Hindole-5-carboxylic acid. Parallel formation of dimers, such as 2,3-dihydro-1H,1'H-2,3'-biindole-5,5'-dicarboxylic acid and trimers, such as 3,3'-[2-(2-amino-5-carboxyphenyl) ethane-1,1-diyl]bis(1H-indole-5-carboxylic acid) of the indole derivatives was also observed. Reaction of a mixture of indole and indole-5-carboxylic acid with 2-phenylethanethiol proceeded in a regioselective way, affording 3-[2-(2-aminophenyl)-1-(phenethylthio)ethyl]-1H-indole-5-carboxylic acid. An additional product of this reaction was 3-[2-(2-aminophenyl)-1-(phenethylthio)ethyl]-2,3-dihydro-1H,1'H-2,3'-biindole-5'-carboxylic acid, which upon standing in DMSO-d6 solution gave 3-[2-(2-aminophenyl)-1-(phenethylthio)ethyl]-1H,1'H-2,3'-biindole-5'-carboxylic acid. Structures of all compounds were elucidated by NMR, and a mechanism for their formation was suggested.

Felikss Mutulis; Adolf Gogoll; Ilze Mutule; Sviatlana Yahorava; Aleh Yahorau; Edvards Liepinsh; Jarl E.S. Wikberg

2008-01-01

198

Thiol-ene/acrylate substrates for softening intracortical electrodes.  

UK PubMed Central (United Kingdom)

Neural interfaces have traditionally been fabricated on rigid and planar substrates, including silicon and engineering thermoplastics. However, the neural tissue with which these devices interact is both 3D and highly compliant. The mechanical mismatch at the biotic-abiotic interface is expected to contribute to the tissue response that limits chronic signal recording and stimulation. In this work, novel ternary thiol-ene/acrylate polymer networks are used to create softening substrates for neural recording electrodes. Thermomechanical properties of the substrates are studied through differential scanning calorimetry and dynamic mechanical analysis both before and after exposure physiological conditions. This substrate system softens from more than 1 GPa to 18 MPa on exposure to physiological conditions: reaching body temperature and taking up less than 3% fluid. The impedance of 177 µm(2) gold electrodes electroplated with platinum black fabricated on these substrates is measured to be 206 k? at 1 kHz. Specifically, intracortical electrodes are fabricated, implanted, and used to record driven neural activity. This work describes the first substrate system that can use the full capabilities of photolithography, respond to physiological conditions by softening markedly after insertion, and record driven neural activity for 4 weeks. © 2013 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2013.

Ware T; Simon D; Liu C; Musa T; Vasudevan S; Sloan A; Keefer EW; Rennaker RL 2nd; Voit W

2013-05-01

199

(Processing and targeting of the thiol protease aleurain)  

Energy Technology Data Exchange (ETDEWEB)

Our goal for work during the past two years under this Grant was to characterize the barley thiol protease, aleurain, to determine if it is secreted or retained intracellularly in aleurone cells, and to begin to elucidate structural features that might control targeting of the protein to its final destination. We have shown that aleurain is synthesized as a proenzyme with two N-linked oligosaccharide chains, one high mannose-type and one complex-type. Aleurain undergoes processing to mature form by removal of an Nterminal prosegment, and is retained intracellularly; it cannot be detected among proteins secreted from aleurone cells. Treatment of aleurone cells with tunicamycin to prevent glycosylation of aleurain does not prevent processing of the unglycosylated form. The N-terminal portion of aleurain's prosegment is homologous to the comparable region in two yeast vacuolar proteases, where that region is known to contain the signal necessary for targeting the proteases to the vacuole. 18 refs., 7 figs.

Rogers, J.C.

1990-01-01

200

Processing and targeting of the thiol protease, aleurain  

Energy Technology Data Exchange (ETDEWEB)

We have identified a cDNA clone from barley aleurone mRNA that encodes a protein with unusual homologies: the C-terminal portion, about 270 amino acids, is 65% identical to the mammalian thiol protease, cathepsin H. This degree of sequence conservation indicates that the enzyme must have some specific function in both plants and mammals that cannot tolerate further divergence. The N-terminal 1/3 of the protein, about 140 amino acids, has no detectable homologies to other known protein sequences; its function is unknown. In aleurone tissue, the mRNA level is increased by gibberellic acid and decreased by abscisic acid, but is expressed apparently constitutively at high levels in leaf and root tissues. The amino acid sequence and cathepsin H homology suggest that the protein will be both secreted into the endoplasmic reticulum and glycosylated. Using our cDNA clone in a bacterial expression system, we have made a fusion protein containing the protease domain of aleurain, and have used it to raise specific antisera in rabbits. These antibodies identify a 32 kd protein in extracts of aleurone layers that is induced with GA treatment but not secreted; a similarly sized protein is specifically identified in extracts of leaf tissue. Experiments are underway to characterize the pattern of expression in different tissues, to identify the subcellular locations of the protein, to characterize processing of the precursor to the 32 kd mature form, and to purify the enzyme from barley. 2 figs.

Rogers, J.C.

1989-01-01

 
 
 
 
201

Control of dihydrofolate reductase messenger ribonucleic acid production  

Energy Technology Data Exchange (ETDEWEB)

The authors used methotrexate-resistant mouse cells in which dihydrofolate reductase levels are approximately 500 times normal to study the effect of growth stimulation on dihydrofolate reductase gene expression. As a result of growth stimulation, the relative rate of dihydrofolate reductase protein synthesis increased threefold, reaching a maximum between 25 and 30 h after stimulation. The relative rate of dihydrofolate reductase messenger ribonucleic acid production (i.e., the appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm) increased threefold after growth stimulation and was accompanied by a corresponding increase in the relative steady-state level of dihydrofolate reductase ribonucleic acid in the nucleus. However, the increase in the nuclear level of dihydrofolate reductase ribonucleic acid was not accompanied by a significant increase in the relative rate of transcription of the dihydrofolate reductase genes. These data indicated that the relative rate of appearance of dihydrofolate reductase messenger ribonucleic acid in the cytoplasm depends on the relative stability of the dihydrofolate reductase ribonucleic acid sequences in the nucleus and is not dependent on the relative rate of transcription of the dihydrofolate reductase genes.

Leys, E.J.; Kellems, R.E.

1981-11-01

202

Facile Synthesis of Thiol-Functionalized Long-Chain Highly Branched ROMP Polymers and Surface-Decorated with Gold Nanoparticles.  

UK PubMed Central (United Kingdom)

The synthesis of thiol-functionalized long-chain highly branched polymers (LCHBPs) has been accomplished in combination of ring-opening metathesis polymerization (ROMP) and thiol-Michael addition click reaction. A monotelechelic polymer with a terminal acrylate and many pendent thiol groups is first prepared through adding an internal cis-olefin terminating agent to the reaction mixture immediately after the completion of the living ROMP, and then utilized as an ABn -type macromonomer in subsequent thiol-ene reaction between acrylate and thiol, yielding LCHBPs as the reaction time prolonged. Au nanoparticles are then covalently conjugated onto the surface of thiol-functionalized LCHBP to fabricate novel hybrid nanostructures, which is shown as one interesting application of such functionalized metathesis polymers. This facile approach can be extended toward the fabrication of novel nanomaterials with sophisticated structures and tunable multifunctionalities.

Ding L; Qiu J; Zhu Z

2013-09-01

203

Substituent Effects on the Reactivity of Benzoquinone Derivatives with Thiols.  

UK PubMed Central (United Kingdom)

Benzoquinone (BQ) is an extremely potent electrophilic contact allergen that haptenates endogenous proteins through Michael addition (MA). It is also hypothesized that BQ may haptenate proteins via free radical formation. The objective of this study was to assess the inductive effects (activating and deactivating) of substituents on BQ reactivity and the mechanistic pathway of covalent binding to a nucleophilic thiol. The BQ binding of Cys34 on human serum albumin was studied, and for reactivity studies, nitrobenzenethiol (NBT) was used as a surrogate for protein binding of the BQ and benzoquinone derivatives (BQD). Stopped flow techniques were used to determine pseudofirst order rate constants (k) of methyl-, t-butyl-, and chlorine-substituted BQD reactions with NBT, whereas electron pair resonance (EPR) studies were performed to investigate the presence of the free radical mediated binding mechanism of BQD. Characterization of adducts was performed using mass spectrometry and nuclear magnetic resonance spectroscopy (NMR). The rate constant values demonstrated the chlorine-substituted (activated) BQD to be more reactive toward NBT than the methyl and t-butyl-substituted (deactivated) BQD, and this correlated with the respective EPR intensities. The EPR signal, however, was quenched in the presence of NBT suggesting MA as the dominant reaction pathway. MS and NMR results confirmed adduct formation to be a result of MA onto the BQ ring with vinylic substitution also occurring for chlorine-substituted derivatives. The binding positions on BQ and NBT/BQ(D) stoichiometric ratios were affected by whether the inductive effects of the substituents on the ring were positive or negative. The reactivity of BQ and BQD is discussed in terms of the potential relationship to potential allergenic potency.

Mbiya W; Chipinda I; Siegel PD; Mhike M; Simoyi RH

2012-12-01

204

Infection free titanium alloys by stabile thiol based nanocoating.  

Science.gov (United States)

As biomedical materials, titanium and titanium alloys (Ti-6Al-4V) are superior to many materials in terms of mechanical properties and biocompatibility. However, they are still not sufficient for prolonged clinical use because the biocompatibility of these materials must be improved. In this study, the prevention of the attachment of test microorganism on the Ti alloy surfaces by thiol (-SH) and hydroxyl (-OH) functional group containing monomer in plasma based electron beam generator was reported in order to prepare anti-fouling surfaces. The precursor, 11-mercaptoundecanoic acid is used as plasma source to create nano-film with 30-60 nm approximately. The surface chemistry and topology of uncoated and coated samples are characterized by Fourier Transform Infrared Spectroscopy (FTIR) and Atomic Force Microscopy (AFM). Static contact angle measurements are performed to state the change of surface hydrophilicity. All coated samples are tested in-vitro environment with Staphylococcus epidermidis that is chosen as the test bacteria strain in view of its significance for the pathogenesis of medical-device-related infections. This test is repeated after certain period of times and samples are waited in dynamic fluid media in order to investigate the stability of nano-coating. Plasma polymerized 11-mercaptoundecanoic acid film (PP MUA) with 42 +/- 4 nm is found alternative, stabile and simple method to create bacterial anti-fouling surfaces. The static contact angle of the coated surface is 34 +/- 80 whereas the uncoated surface is 57 +/- 50. For the coated surface, the presence of C-OH and C==O groups in infrared spectra defining the PP MUA is achieved by the plasma polymerization. The attachment of the model microorganism on the biomaterial surface prepared by PP MUA is reduced 85.3% if compared to unmodified control surface. PMID:20355467

Cökeliler, Dilek; Gökta?, Hilal; Tosun, Pinar Deniz; Mutlu, Selma

2010-04-01

205

Infection free titanium alloys by stabile thiol based nanocoating.  

UK PubMed Central (United Kingdom)

As biomedical materials, titanium and titanium alloys (Ti-6Al-4V) are superior to many materials in terms of mechanical properties and biocompatibility. However, they are still not sufficient for prolonged clinical use because the biocompatibility of these materials must be improved. In this study, the prevention of the attachment of test microorganism on the Ti alloy surfaces by thiol (-SH) and hydroxyl (-OH) functional group containing monomer in plasma based electron beam generator was reported in order to prepare anti-fouling surfaces. The precursor, 11-mercaptoundecanoic acid is used as plasma source to create nano-film with 30-60 nm approximately. The surface chemistry and topology of uncoated and coated samples are characterized by Fourier Transform Infrared Spectroscopy (FTIR) and Atomic Force Microscopy (AFM). Static contact angle measurements are performed to state the change of surface hydrophilicity. All coated samples are tested in-vitro environment with Staphylococcus epidermidis that is chosen as the test bacteria strain in view of its significance for the pathogenesis of medical-device-related infections. This test is repeated after certain period of times and samples are waited in dynamic fluid media in order to investigate the stability of nano-coating. Plasma polymerized 11-mercaptoundecanoic acid film (PP MUA) with 42 +/- 4 nm is found alternative, stabile and simple method to create bacterial anti-fouling surfaces. The static contact angle of the coated surface is 34 +/- 80 whereas the uncoated surface is 57 +/- 50. For the coated surface, the presence of C-OH and C==O groups in infrared spectra defining the PP MUA is achieved by the plasma polymerization. The attachment of the model microorganism on the biomaterial surface prepared by PP MUA is reduced 85.3% if compared to unmodified control surface.

Cökeliler D; Gökta? H; Tosun PD; Mutlu S

2010-04-01

206

Electrochemistry behavior of endogenous thiols on fluorine doped tin oxide electrodes  

Energy Technology Data Exchange (ETDEWEB)

Highlights: > The first time that fluorine doped tin oxide electrodes are used for the electrooxidation of endogenous thiols. > Low potentials of electrooxidation were obtained for the different thiols. > The electrochemical behavior of thiols depends on the pH and the ionic electroactive species, the electrooxidation proceeds for a process of adsorption of electroactive species on FTO and high values the heterogeneous electron tranfer rate constant of the reaction were obtained. - Abstract: In this work the electrochemical behavior of different thiols on fluorine doped tin oxide (FTO) electrodes is reported. To this end, the mechanism of electrochemical oxidation of glutathione (GSH), cysteine (Cys), homocysteine (HCys) and acetyl-cysteine (ACys) at different pH was investigated. FTO showed electroactivity for the oxidation of the first three thiols at pH between 2.0 and 4.0, but under these conditions no acetyl-cysteine oxidation was observed on FTO. Voltammetric studies of the electro-oxidation of GSH, Cys and HCys showed peaks at about 0.35, 0.29, and 0.28 V at optimum pH 2.4, 2.8 and 3.4, respectively. In addition, this study demonstrated that GSH, Cys and HCys oxidation occurs when the zwitterion is the electro-active species that interact by adsorption on FTO electrodes. The overall reaction involves 4e{sup -}/4H{sup +} and 2e{sup -}/2H{sup +}, respectively, for HCys and for GSH and Cys and high heterogeneous electron transfer rate constants. Besides, the use of FTO for the determination of different thiols was evaluated. Experimental square wave voltammetry shows a linear current vs. concentrations response between 0.1 and 1.0 mM was found for HCys and GSH, indicating that these FTO electrodes are promising candidates for the efficient electrochemical determination of these endogenous thiols.

Rojas, Luciana; Molero, Leonard; Tapia, Ricardo A.; Rio, Rodrigo del; Valle, M. Angelica del; Antilen, Monica [Departamento de Quimica Inorganica, Facultad de Quimica, Pontificia Universidad Catolica de Chile, Av Vicuna Mackenna 4860, Casilla 306, Correo 22, Macul, Santiago (Chile); Armijo, Francisco, E-mail: jarmijom@uc.cl [Departamento de Quimica Inorganica, Facultad de Quimica, Pontificia Universidad Catolica de Chile, Av Vicuna Mackenna 4860, Casilla 306, Correo 22, Macul, Santiago (Chile)

2011-10-01

207

Electrochemistry behavior of endogenous thiols on fluorine doped tin oxide electrodes  

International Nuclear Information System (INIS)

[en] Highlights: ? The first time that fluorine doped tin oxide electrodes are used for the electrooxidation of endogenous thiols. ? Low potentials of electrooxidation were obtained for the different thiols. ? The electrochemical behavior of thiols depends on the pH and the ionic electroactive species, the electrooxidation proceeds for a process of adsorption of electroactive species on FTO and high values the heterogeneous electron tranfer rate constant of the reaction were obtained. - Abstract: In this work the electrochemical behavior of different thiols on fluorine doped tin oxide (FTO) electrodes is reported. To this end, the mechanism of electrochemical oxidation of glutathione (GSH), cysteine (Cys), homocysteine (HCys) and acetyl-cysteine (ACys) at different pH was investigated. FTO showed electroactivity for the oxidation of the first three thiols at pH between 2.0 and 4.0, but under these conditions no acetyl-cysteine oxidation was observed on FTO. Voltammetric studies of the electro-oxidation of GSH, Cys and HCys showed peaks at about 0.35, 0.29, and 0.28 V at optimum pH 2.4, 2.8 and 3.4, respectively. In addition, this study demonstrated that GSH, Cys and HCys oxidation occurs when the zwitterion is the electro-active species that interact by adsorption on FTO electrodes. The overall reaction involves 4e-/4H+ and 2e-/2H+, respectively, for HCys and for GSH and Cys and high heterogeneous electron transfer rate constants. Besides, the use of FTO for the determination of different thiols was evaluated. Experimental square wave voltammetry shows a linear current vs. concentrations response between 0.1 and 1.0 mM was found for HCys and GSH, indicating that these FTO electrodes are promising candidates for the efficient electrochemical determination of these endogenous thiols.

2011-10-01

208

Thiol involvement in the inhibition of DNA repair by metals in mammalian cells  

International Nuclear Information System (INIS)

We have previously demonstrated that a number of metal salts have the capacity to inhibit the DNA repair process in human cells. We investigated repair of X-ray damage in metal-treated HeLa cells under normal conditions and conditions in which cellular thiols had been depleted by treatment with buthionine sulfoximine (BSO) and diethyl maleate (DEM). The combination reduced cellular TNPT by 92%, and cells so depleted became sensitized to X-ray-induced killing and exhibited retarded sealing of X-ray-induced DNA single-strand breaks. Thiol depletion also sensitized cells to the cytotoxicity of certain but not all metals tested. The sensitivity to copper was increased over 6000-fold, and significant enhancement of killing was also seen in cells treated with arsenic, lead, and mercury. Smaller effects were observed with cadmium and nickel, and sensitivity to manganese, magnesium, cobalt or zinc was not substantially altered. Enhanced sensitivity to X-ray killing was found in cells treated with nickel, cadmium, zinc, arsenic, and copper under conditions in which thiols were not limiting. In thiol-depleted cells, sensitivity was not further increased in the case of nickel and arsenic but at least additively affected for copper, mercury and zinc. X-Ray-induced single-strand break repair was retarded by treatment of cells with mercury, nickel, zinc, arsenic, and copper in thiol-normal cells. In thiol-depleted cells, repair inhibition by zinc, arsenic, and copper was nearly complete, while little additional effect on repair was seen following mercury and nickel treatment. An examination of the effects of brief metal treatment on cellular TNPT revealed that copper strongly decreased thiol levels whereas the other metals tested either had no effect on TNPT or reduced TNPT levels to no less than 48% under the conditions employed.

1989-01-01

209

Lapachol inhibition of vitamin K epoxide reductase and vitamin K quinone reductase.  

UK PubMed Central (United Kingdom)

Lapachol [2-hydroxy-3-(3-methyl-2-butenyl)-1,4-naphthoquinone] has been shown to be a potent inhibitor of both vitamin K epoxide reductase and the dithiothreitol-dependent vitamin K quinone reductase of rat liver microsomes in vitro. These observations explain the anticoagulant activity of lapachol previously observed in both rats and humans. Lapachol inhibition of the vitamin K epoxide and quinone reductases resembled coumarin anticoagulant inhibition, and was observed in normal strain but not in warfarin-resistant strain rat liver microsomes. This similarity of action suggests that the lactone functionality of the coumarins is not critical for their activity. The initial-velocity steady-state inhibition patterns for lapachol inhibition of the solubilized vitamin K epoxide reductase were consistent with tight binding of lapachol to the oxidized form of the enzyme, and somewhat lower affinity for the reduced form. It is proposed that lapachol assumes a 4-enol tautomeric structure similar to that of the 4-hydroxy coumarins. These structures are analogs of the postulated hydroxyvitamin K enolate intermediate bound to the oxidized form of the enzyme in the chemical reaction mechanism of vitamin K epoxide reductase, thus explaining their high affinity.

Preusch PC; Suttie JW

1984-11-01

210

Mechanism of action of clostridial glycine reductase: Isolation and characterization of a covalent acetyl enzyme intermediate  

International Nuclear Information System (INIS)

[en] Clostridial glycine reductase consists of proteins A, B, and C and catalyzes the reaction glycine + Pi + 2e- ? acetyl phosphate + NH4+. Evidence was previously obtained that is consistent with the involvement of an acyl enzyme intermediate in this reaction. The authors now demonstrate that protein C catalyzes exchange of [32P]Pi into acetyl phosphate, providing additional support for an acetyl enzyme intermediate on protein C. Furthermore, they have isolated acetyl protein C and shown that it is qualitatively, catalytically competent. Acetyl protein C can be obtained through the forward reaction from protein C and Se-(carboxymethyl)selenocysteine-protein A, which is generated by the reaction of glycine with proteins A and B. Acetyl protein C can also be generated through the reverse reaction by the addition of acetyl phosphate to protein C. Both procedures lead to the same acetyl enzyme. The acetyl enzyme reacts with Pi to give acetyl phosphate. When [14C]acetyl protein C is denaturated with TCA and redissolved with urea, radioactivity remained associated with the protein. Treatment with KBH4 removes all the radioactivity associated with protein C, resulting in the formation of [14C]ethanol. They conclude that a thiol group on protein C is acetylated. Proteins A and C together catalyze the exchange of tritium atoms from [3H]H2O into acetyl phosphate. This exchange reaction supports the proposal that an enol of the acetyl enzyme is an intermediate in the reaction sequence

1991-04-23

211

The sulfur shift: an activation mechanism for periplasmic nitrate reductase and formate dehydrogenase.  

UK PubMed Central (United Kingdom)

A structural rearrangement known as sulfur shift occurs in some Mo-containing enzymes of the DMSO reductase family. This mechanism is characterized by the displacement of a coordinating cysteine thiol (or SeCys in Fdh) from the first to the second shell of the Mo-coordination sphere metal. The hexa-coordinated Mo ion found in the as-isolated state cannot bind directly any exogenous ligand (substrate or inhibitors), while the penta-coordinated ion, attained upon sulfur shift, has a free binding site for direct coordination of the substrate. This rearrangement provides an efficient mechanism to keep a constant coordination number throughout an entire catalytic pathway. This mechanism is very similar to the carboxylate shift observed in Zn-dependent enzymes, and it has been recently detected by experimental means. In the present paper, we calculated the geometries and energies involved in the sulfur-shift mechanism using QM-methods (M06/(6-311++G(3df,2pd),SDD)//B3LYP/(6-31G(d),SDD)). The results indicated that the sulfur-shift mechanism provides an efficient way to enable the metal ion for substrate coordination.

Cerqueira NM; Fernandes PA; Gonzalez PJ; Moura JJ; Ramos MJ

2013-10-01

212

Investigation of reactions postulated to occur during inhibition of ribonucleotide reductases by 2'-azido-2'-deoxynucleotides.  

UK PubMed Central (United Kingdom)

Model 3'-azido-3'-deoxynucleosides with thiol or vicinal dithiol substituents at C2' or C5' were synthesized to study reactions postulated to occur during inhibition of ribonucleotide reductases by 2'-azido-2'-deoxynucleotides. Esterification of 5'-(tert-butyldiphenylsilyl)-3'-azido-3'-deoxyadenosine and 3'-azido-3'-deoxythymidine (AZT) with 2,3-S-isopropylidene-2,3-dimercaptopropanoic acid or N-Boc-S-trityl-L-cysteine and deprotection gave 3'-azido-3'-deoxy-2'-O-(2,3-dimercaptopropanoyl or cysteinyl)adenosine and the 3'-azido-3'-deoxy-5'-O-(2,3-dimercaptopropanoyl or cysteinyl)thymidine analogs. Density functional calculations predicted that intramolecular reactions between generated thiyl radicals and an azido group on such model compounds would be exothermic by 33.6-41.2 kcal/mol and have low energy barriers of 10.4-13.5 kcal/mol. Reduction of the azido group occurred to give 3'-amino-3'-deoxythymidine, which was postulated to occur with thiyl radicals generated by treatment of 3'-azido-3'-deoxy-5'-O-(2,3-dimercaptopropanoyl)thymidine with 2,2'-azobis-(2-methyl-2-propionamidine) dihydrochloride. Gamma radiolysis of N(2)O-saturated aqueous solutions of AZT and cysteine produced 3'-amino-3'-deoxythymidine and thymine most likely by both radical and ionic processes.

Dang TP; Sobczak AJ; Mebel AM; Chatgilialoglu C; Wnuk SF

2012-07-01

213

Investigation of reactions postulated to occur during inhibition of ribonucleotide reductases by 2'-azido-2'-deoxynucleotides.  

Science.gov (United States)

Model 3'-azido-3'-deoxynucleosides with thiol or vicinal dithiol substituents at C2' or C5' were synthesized to study reactions postulated to occur during inhibition of ribonucleotide reductases by 2'-azido-2'-deoxynucleotides. Esterification of 5'-(tert-butyldiphenylsilyl)-3'-azido-3'-deoxyadenosine and 3'-azido-3'-deoxythymidine (AZT) with 2,3-S-isopropylidene-2,3-dimercaptopropanoic acid or N-Boc-S-trityl-L-cysteine and deprotection gave 3'-azido-3'-deoxy-2'-O-(2,3-dimercaptopropanoyl or cysteinyl)adenosine and the 3'-azido-3'-deoxy-5'-O-(2,3-dimercaptopropanoyl or cysteinyl)thymidine analogs. Density functional calculations predicted that intramolecular reactions between generated thiyl radicals and an azido group on such model compounds would be exothermic by 33.6-41.2 kcal/mol and have low energy barriers of 10.4-13.5 kcal/mol. Reduction of the azido group occurred to give 3'-amino-3'-deoxythymidine, which was postulated to occur with thiyl radicals generated by treatment of 3'-azido-3'-deoxy-5'-O-(2,3-dimercaptopropanoyl)thymidine with 2,2'-azobis-(2-methyl-2-propionamidine) dihydrochloride. Gamma radiolysis of N(2)O-saturated aqueous solutions of AZT and cysteine produced 3'-amino-3'-deoxythymidine and thymine most likely by both radical and ionic processes. PMID:22711937

Dang, Thao P; Sobczak, Adam J; Mebel, Alexander M; Chatgilialoglu, Chryssostomos; Wnuk, Stanislaw F

2012-04-21

214

Nebulized and oral thiol derivatives for pulmonary disease in cystic fibrosis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Cystic fibrosis is an inherited condition resulting in thickened, sticky respiratory secretions. Respiratory failure, due to recurrent pulmonary infection and inflammation, is the most common cause of mortality. Muco-active therapies (e.g. dornase alfa and nebulized hypertonic saline) may decrease sputum viscosity, increase airway clearance of sputum, reduce infection and inflammation and improve lung function. Thiol derivatives, either oral or nebulized, have shown benefit in other respiratory diseases. Their mode of action is likely to differ according to the route of administration. There are several thiol derivatives, and it is unclear which of these may be beneficial in cystic fibrosis. OBJECTIVES: To evaluate the efficacy and safety of nebulized and oral thiol derivatives in people with cystic fibrosis. SEARCH METHODS: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register, comprising references identified from comprehensive electronic database searches, hand searches of relevant journals, abstract books and conference proceedings.Most recent search: 13 June 2013.We also conducted a PubMed search on 26 February 2013 for relevant published articles. SELECTION CRITERIA: Randomized and quasi-randomized controlled trials comparing nebulized or oral thiol derivatives to placebo or another thiol derivative in people with cystic fibrosis. DATA COLLECTION AND ANALYSIS: The authors independently assessed trials for inclusion, analysed risk of bias and extracted data. MAIN RESULTS: Searches identified 23 trials; nine trials (255 participants) are included, of these seven trials are more than 10 years old. Three trials of nebulized thiol derivatives were identified (one compared 20% N-acetylcysteine to 2% N-acetylcysteine; another compared sodium-2-mercaptoethane sulphonate to 7% hypertonic saline; and another compared glutathione to 4% hypertonic saline). Although generally well-tolerated with no significant adverse effects, there was no evidence of significant clinical benefit in our primary outcomes in participants receiving these treatments.Six trials of oral thiol derivatives were identified. Three trials compared N-acetylcysteine to placebo; one compared N-acetylcysteine, ambroxol and placebo; one compared carbocysteine to ambroxol; and one compared low and high-dose N-acetylcysteine. Oral thiol derivatives were generally well-tolerated with no significant adverse effects, however there was no evidence of significant clinical benefit in our primary outcomes in participants receiving these treatments. AUTHORS' CONCLUSIONS: We found no evidence to recommend the use of either nebulized or oral thiol derivatives in people with cystic fibrosis. There are very few good quality trials investigating the effect of these medications in cystic fibrosis, and further research is required to investigate the potential role of these medications in improving the outcomes of people with cystic fibrosis.

Tam J; Nash EF; Ratjen F; Tullis E; Stephenson A

2013-01-01

215

Nebulized and oral thiol derivatives for pulmonary disease in cystic fibrosis.  

UK PubMed Central (United Kingdom)

BACKGROUND: Cystic fibrosis is an inherited condition resulting in thickened, sticky respiratory secretions. Respiratory failure, due to recurrent pulmonary infection and inflammation, is the most common cause of mortality. Muco-active therapies (e.g. dornase alfa and nebulized hypertonic saline) may decrease sputum viscosity, increase airway clearance of sputum, reduce infection and inflammation and improve lung function. Thiol derivatives, either oral or nebulized, have shown benefit in other respiratory diseases. Their mode of action is likely to differ according to the route of administration. There are several thiol derivatives, and it is unclear which of these may be beneficial in cystic fibrosis. OBJECTIVES: To evaluate the efficacy and safety of nebulized and oral thiol derivatives in people with cystic fibrosis. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group Trials Register, comprising references identified from comprehensive electronic database searches, hand searches of relevant journals, abstract books and conference proceedings.Most recent search: November 2008. SELECTION CRITERIA: Randomized and quasi-randomized controlled trials comparing nebulized or oral thiol derivatives to placebo or another thiol derivative in people with cystic fibrosis. DATA COLLECTION AND ANALYSIS: The authors independently assessed trials for inclusion, analysed methodological quality and extracted data. MAIN RESULTS: Searches identified 18 trials; eight (seven older than 10 years) (234 participants) are included. Three trials of nebulized thiol derivatives were identified (one compared 20% n-acetylcysteine to 2% n-acetylcysteine; another compared sodium-2-mercaptoethane sulphonate to 7% hypertonic saline; and another compared glutathione to 4% hypertonic saline). Although generally well-tolerated with no significant adverse effects, there was no evidence of significant clinical benefit in our primary outcomes in participants receiving these treatments.Five studies of oral thiol derivatives were identified. Three studies compared n-acetylcysteine to placebo; one compared n-acetylcysteine, ambroxol and placebo; and one compared carbocysteine to ambroxol. Oral thiol derivatives were generally well-tolerated with no significant adverse effects, however there was no evidence of significant clinical benefit in our primary outcomes in participants receiving these treatments. AUTHORS' CONCLUSIONS: We found no evidence to recommend the use of either nebulized or oral thiol derivatives in people with cystic fibrosis. There are very few good quality trials investigating the effect of these medications in cystic fibrosis, and further research is required to investigate the potential role of these medications in improving the outcomes of people with cystic fibrosis.

Nash EF; Stephenson A; Ratjen F; Tullis E

2009-01-01

216

General anesthesia and methylenetetrahydrofolate reductase deficiency.  

UK PubMed Central (United Kingdom)

Methylenetetrahydrofolate reductase (MTHFR) deficiency is an autosomal recessive disorder with a spectrum of manifestations including neurological symptoms, premature arteriosclerosis, and venous and arterial thrombosis. Most patients are heterozygous for multiple MTHFR substitutions; small minorities are homozygous for mutations at this locus. Among these mutations, the C677T polymorphism is the most deleterious. Nitrous oxide use in anesthesia leads to significant increases in plasma homocysteine. We present a patient undergoing urgent surgery with a preoperative diagnosis of homozygous MTHFR deficiency.

Shay H; Frumento RJ; Bastien A

2007-01-01

217

cDNA FOR HUMAN METHYLENETETRAHYDROFOLATE REDUCTASE  

UK PubMed Central (United Kingdom)

The present invention relates to a cDNA probe for human methylenetetrahydrofolate reductase (MTHFR), and its uses. The probe of the present invention may be used for the identification of sequence abnormalities in patients with severe or mild MTHFR deficiency, including cardiovascular patients and patients with neurologic symptoms. A human MTHFR protein which hybridizes to the probe of the present invention may be used for therapy of MTHFR-deficiency patients by biochemical or pharmacological approaches.

ROZEN Rima; GOYETTE Philippe

218

General anesthesia and methylenetetrahydrofolate reductase deficiency.  

Science.gov (United States)

Methylenetetrahydrofolate reductase (MTHFR) deficiency is an autosomal recessive disorder with a spectrum of manifestations including neurological symptoms, premature arteriosclerosis, and venous and arterial thrombosis. Most patients are heterozygous for multiple MTHFR substitutions; small minorities are homozygous for mutations at this locus. Among these mutations, the C677T polymorphism is the most deleterious. Nitrous oxide use in anesthesia leads to significant increases in plasma homocysteine. We present a patient undergoing urgent surgery with a preoperative diagnosis of homozygous MTHFR deficiency. PMID:18008117

Shay, Hamilton; Frumento, Robert J; Bastien, Alexandra

2007-11-01

219

cDNA for human methylenetetrahydrofolate reductase  

UK PubMed Central (United Kingdom)

The present invention relates to a cDNA probe for human methylenetetrahydrofolate reductase (MTHFR), and its uses, The probe of the present invention may be used for the identification of sequence abnormalities in patients with severe or mild MTHFR deficiency, including cardiovascular patients and patients with neurologic symptoms, A human MTHFR protein which hybridizes to the probe of the present invention may be used for therapy of MTHFR-deficiency patients by biochemical or pharmacological approaches.

ROZEN RIMA; GOYETTE PHILIPPE

220

Improving the reliability of human serum albumin-thiol group determination.  

UK PubMed Central (United Kingdom)

The thiol (Cys34) content of human serum albumin (HSA-SH) decreases during oxidative and carbonyl stress and, therefore, could represent a useful parameter in clinical practice. Nevertheless, the reliability of HSA-thiol determination with Ellman's method depends on the purity of isolated HSA. Determination of total serum thiols (mmol/L) and HSA-SH content (mmol -SH/mmol HSA) after HSA isolation from diabetic patient and control sera by a two-step precipitation with ammonium sulfate (AS), as well as HSA-SH contribution (%) to total serum thiols, was assessed. Purity and yield of isolated HSA were monitored spectrophotometrically and by native polyacrylamide gel electrophoresis. Precipitation of HSA from serum via a two-step method with AS produced HSA with 91.9±3.6% purity and 69.7±4.4% yield, allowing for precise (relative standard deviation of 3.2%) and reliable (comparing with total serum thiols) measurement of HSA-SH content with DTNB [5,5'-dithiobis-(2-nitrobenzoic acid)]. The content of the HSA-SH group in patients with type 2 diabetes was significantly (P<0.05) lower compared with that of the healthy cohort (0.483±0.067 vs. 0.561±0.054 mmol -SH/mmol HSA). Because the proposed method of HSA isolation is simple, time-efficient, and technically less demanding, and it also enables reliable determination of HSA-SH content, it is suitable for clinical practice.

Jovanovi? VB; Penezi?-Romanjuk AZ; Pavi?evi? ID; A?imovi? JM; Mandi? LM

2013-08-01

 
 
 
 
221

Influence of thiol stress on oxidative phosphorylation and generation of ROS in Streptomyces coelicolor  

Directory of Open Access Journals (Sweden)

Full Text Available Thiols play very important role in the intracellular redox homeostasis. Imbalance in the redox status leads to changes in the intracellular metabolism including respiration. Thiol stress, a reductive type of stress can also cause redox imbalance. When Gram-positive bacterium Strep- tomyces coelicolor was exposed to thiol stress, catalaseA was induced. Induction of catalaseA is the consequence of elevation of ROS (reactive oxygen species). The two major sources of reactive oxygen species are Fenton reaction and slippage of electrons from electron transport chain during respiration. Hence, the effect of thiol stress was checked on the rate of oxidative phosphorylation in S. coelicolor. We found correlation in the increase of oxidative phosphorylation rate and the generation of ROS, subsequently leading to induction of catalase. It was observed that thiol stress does not affect the functionality of the individual complexes of the ETC, but still there was an increase in the overall respiration, which may lead to generation of more ROS leading to induction of catalase.

Hemendra J. Vekaria; Ratna Prabha Chivukula

2010-01-01

222

Quantification of protein thiols using ThioGlo 1 fluorescent derivatives and HPLC separation.  

UK PubMed Central (United Kingdom)

A method for quantification of total soluble protein-derived thiols in beer was developed based on the formation of fluorescent adducts with the maleimide compound ThioGlo 1. The problem of interference from fluorescent adducts of sulfite and ThioGlo 1 was solved by HPLC separation of the adducts followed by fluorescence detection. Using standard addition of GSH, a detection limit of 0.028 ?M thiols was achieved. The application and validation of the method was demonstrated for beers with different color intensities, and the application range is in principle for any biological system containing thiols. However, the quantification of cysteine was complicated by a lower fluorescence response of its ThioGlo 1 adducts. Based on the studies of the responses of a series of cysteine-derived thiols and (1)H NMR studies of the structures of ThioGlo 1 adducts with GSH and cysteine, it was concluded that thiols with a neighboring free amino group yield ThioGlo 1 adducts with a reduced fluorescence intensity.

Hoff S; Larsen FH; Andersen ML; Lund MN

2013-04-01

223

Oxidation of the albumin thiol to sulfenic acid and its implications in the intravascular compartment  

Scientific Electronic Library Online (English)

Full Text Available Abstract in english Human serum albumin (HSA) is the most abundant protein in the intravascular compartment. It possesses a single thiol, Cys34, which constitutes ~80% of the total thiols in plasma. This thiol is able to scavenge plasma oxidants. A central intermediate in this potential antioxidant activity of human serum albumin is sulfenic acid (HSA-SOH). Work from our laboratories has demonstrated the formation of a relatively stable sulfenic acid in albumin through complementary spectrop (more) hotometric and mass spectrometric approaches. Recently, we have been able to obtain quantitative data that allowed us to measure the rate constants of sulfenic acid reactions with molecules of analytical and biological interest. Kinetic considerations led us to conclude that the most likely fate for sulfenic acid formed in the plasma environment is the reaction with low molecular weight thiols to form mixed disulfides, a reversible modification that is actually observed in ~25% of circulating albumin. Another possible fate for sulfenic acid is further oxidation to sulfinic and sulfonic acids. These irreversible modifications are also detected in the circulation. Oxidized forms of albumin are increased in different pathophysiological conditions and sulfenic acid lies in a mechanistic junction, relating oxidizing species to final thiol oxidation products.

Turell, L.; Carballal, S.; Botti, H.; Radi, R.; Alvarez, B.

2009-04-01

224

The electrophilic addition of thiols to olefins: A theoretical and experimental study  

Science.gov (United States)

Density functional theory with the B3LYP hybrid functional and 6-31G* basis set was used to study the geometric and electronic structure of H2C = CHR (R = H, CH3, C2H5, C3H7, C4H9, and C5H11) olefins, their carbocations formed in the addition of the proton to the olefins, R'-S-H aliphatic thiols (R' = H, CH3, C2H5, and C3H7), the products of the addition of thiols to carbocations, and the final products of the addition of thiols to olefins. The proton affinity of the olefins and the products of the addition of thiols to olefins was calculated. The conclusion was drawn that the limiting stage in the nonradical addition of thiols to olefins catalyzed by acids was proton transfer from the protonated reaction product to the olefin. The theoretical results were compared with the experimental data on the electrophilic addition of polymercaptan to heptene-1.

Borisov, Yu. A.; Dyusengaliev, A. K.; Dyusengaliev, K. I.; Serikov, T. P.

2008-12-01

225

A central role for thiols in plant tolerance to abiotic stress.  

UK PubMed Central (United Kingdom)

Abiotic stress poses major problems to agriculture and increasing efforts are being made to understand plant stress response and tolerance mechanisms and to develop new tools that underpin successful agriculture. However, the molecular mechanisms of plant stress tolerance are not fully understood, and the data available is incomplete and sometimes contradictory. Here, we review the significance of protein and non-protein thiol compounds in relation to plant tolerance of abiotic stress. First, the roles of the amino acids cysteine and methionine, are discussed, followed by an extensive discussion of the low-molecular-weight tripeptide, thiol glutathione, which plays a central part in plant stress response and oxidative signalling and of glutathione-related enzymes, including those involved in the biosynthesis of non-protein thiol compounds. Special attention is given to the glutathione redox state, to phytochelatins and to the role of glutathione in the regulation of the cell cycle. The protein thiol section focuses on glutaredoxins and thioredoxins, proteins with oxidoreductase activity, which are involved in protein glutathionylation. The review concludes with a brief overview of and future perspectives for the involvement of plant thiols in abiotic stress tolerance.

Zagorchev L; Seal CE; Kranner I; Odjakova M

2013-01-01

226

A Central Role for Thiols in Plant Tolerance to Abiotic Stress  

Science.gov (United States)

Abiotic stress poses major problems to agriculture and increasing efforts are being made to understand plant stress response and tolerance mechanisms and to develop new tools that underpin successful agriculture. However, the molecular mechanisms of plant stress tolerance are not fully understood, and the data available is incomplete and sometimes contradictory. Here, we review the significance of protein and non-protein thiol compounds in relation to plant tolerance of abiotic stress. First, the roles of the amino acids cysteine and methionine, are discussed, followed by an extensive discussion of the low-molecular-weight tripeptide, thiol glutathione, which plays a central part in plant stress response and oxidative signalling and of glutathione-related enzymes, including those involved in the biosynthesis of non-protein thiol compounds. Special attention is given to the glutathione redox state, to phytochelatins and to the role of glutathione in the regulation of the cell cycle. The protein thiol section focuses on glutaredoxins and thioredoxins, proteins with oxidoreductase activity, which are involved in protein glutathionylation. The review concludes with a brief overview of and future perspectives for the involvement of plant thiols in abiotic stress tolerance.

Zagorchev, Lyuben; Seal, Charlotte E.; Kranner, Ilse; Odjakova, Mariela

2013-01-01

227

Superoxide dismutase and media dependence of far-UV radiation resistance in thiol-treated cells  

International Nuclear Information System (INIS)

Pretreatment of wild-type Escherichia coli K12 cells with dithiothreitol (DTT) induces far-UV radiation resistance after the thiol is removed (Claycamp 1988). The present study shows that a 1 h treatment of cells with DTT in minimal medium followed by a 0.5 h incubation in buffer (370C) results in a dose reduction factor (DRF) calculated at F37 of 1.81. When the thio pretreatment was in rich medium, sensitization occurs with DRF = 0.729. This could be reversed to protection by inhibiting extracellular thiol oxidation in rich medium with the chelator, DETAPAC, such that thiol oxidation rate was equivalent to that of DTT in minimal medium. Both thiol-induced resistance and sensitization produced changes predominantly in the shoulders of survival curves. For either protection or sensitization, at least one form of endogenous superoxide dismutase (SOD) was required. These results suggest that different targets are involved in thiol-induced UV protection and sensitization: DNA and extracellular targets (e.g. the membrane), respectively. (author).

1990-01-01

228

Evaluation of thiol broth for the culture of Salmonella typhi and other bacteria from blood.  

Science.gov (United States)

Thiol broth is known to neutralize various antimicrobial agents. Positivity of growth of various species of bacteria from blood in thiol broth was reported as similar to that in tryptic soy broth (TSB). As blood cultures are often used for the diagnosis of typhoid fever, and as patients may receive antimicrobial therapy before blood culture, the positivity and rapidity of growth of Salmonella typhi in thiol broth were compared to those in TSB. Routine blood culture samples from Yonsei Medical Center patients were inoculated in 50-ml amounts of TSB and thiol broth. The media were prepared from dehydrated products and did not contain CO2, but TSB contained 0.025% sodium polyanethol sulfonate (SPS). Growth of S. paratyphi-A, Klebsiella pneumoniae, Enterobacter sp., Serratia marcescens and alpha-hemolytic Streptococcus were similar in both media. However, greater positivity and shorter incubation time for macroscopic detection were noted in TSB with S. typhi, Escherichia coli and Staphylococcus aureus. It is concluded that thiol broth is inferior to TSB plus SPS for the culture of S. typhi from blood. PMID:2145703

Chong, Y; Kang, M S; Lee, S Y

1990-06-01

229

Evaluation of thiol broth for the culture of Salmonella typhi and other bacteria from blood.  

UK PubMed Central (United Kingdom)

Thiol broth is known to neutralize various antimicrobial agents. Positivity of growth of various species of bacteria from blood in thiol broth was reported as similar to that in tryptic soy broth (TSB). As blood cultures are often used for the diagnosis of typhoid fever, and as patients may receive antimicrobial therapy before blood culture, the positivity and rapidity of growth of Salmonella typhi in thiol broth were compared to those in TSB. Routine blood culture samples from Yonsei Medical Center patients were inoculated in 50-ml amounts of TSB and thiol broth. The media were prepared from dehydrated products and did not contain CO2, but TSB contained 0.025% sodium polyanethol sulfonate (SPS). Growth of S. paratyphi-A, Klebsiella pneumoniae, Enterobacter sp., Serratia marcescens and alpha-hemolytic Streptococcus were similar in both media. However, greater positivity and shorter incubation time for macroscopic detection were noted in TSB with S. typhi, Escherichia coli and Staphylococcus aureus. It is concluded that thiol broth is inferior to TSB plus SPS for the culture of S. typhi from blood.

Chong Y; Kang MS; Lee SY

1990-06-01

230

Comparative proteomic analysis of thiol proteins in the liver after oxidative stress induced by diethylnitrosamine.  

UK PubMed Central (United Kingdom)

Conversion of protein -SH groups to disulfides is an early event during protein oxidation, which has prompted great interest in the study of thiol proteins. Chemical carcinogenesis is strongly associated with the formation of reactive oxygen species (ROS). The goal of this study was to detect thiol proteins that are sensitive to ROS generated during diethylnitrosamine (DEN) metabolism in the rat liver. DEN has been widely used to induce experimental hepatocellular carcinoma. We used modified redox-differential gel electrophoresis (redox-DIGE method) and mass spectrometry MALDI TOF/TOF to identify differential oxidation protein profiles associated with carcinogen exposure. Our analysis revealed a time-dependent increase in the number of oxidized thiol proteins after carcinogen treatment; some of these proteins have antioxidant activity, including thioredoxin, peroxirredoxin 2, peroxiredoxin 6 and glutathione S-transferase alpha-3. According to functional classifications, the identified proteins in our study included chaperones, oxidoreductases, activity isomerases, hydrolases and other protein-binding partners. This study demonstrates that oxidative stress generated by DEN tends to increase gradually through DEN metabolism, causes time-dependent necrosis in the liver and has an oxidative effect on thiol proteins, thereby increasing the number of oxidized thiol proteins. Furthermore, these events occurred during the hepatocarcinogenesis initiation period.

Aparicio-Bautista DI; Pérez-Carreón JI; Gutiérrez-Nájera N; Reyes-Grajeda JP; Arellanes-Robledo J; Vásquez-Garzón VR; Jiménez-García MN; Villa-Treviño S

2013-08-01

231

Oxidation of the albumin thiol to sulfenic acid and its implications in the intravascular compartment  

Directory of Open Access Journals (Sweden)

Full Text Available Human serum albumin (HSA) is the most abundant protein in the intravascular compartment. It possesses a single thiol, Cys34, which constitutes ~80% of the total thiols in plasma. This thiol is able to scavenge plasma oxidants. A central intermediate in this potential antioxidant activity of human serum albumin is sulfenic acid (HSA-SOH). Work from our laboratories has demonstrated the formation of a relatively stable sulfenic acid in albumin through complementary spectrophotometric and mass spectrometric approaches. Recently, we have been able to obtain quantitative data that allowed us to measure the rate constants of sulfenic acid reactions with molecules of analytical and biological interest. Kinetic considerations led us to conclude that the most likely fate for sulfenic acid formed in the plasma environment is the reaction with low molecular weight thiols to form mixed disulfides, a reversible modification that is actually observed in ~25% of circulating albumin. Another possible fate for sulfenic acid is further oxidation to sulfinic and sulfonic acids. These irreversible modifications are also detected in the circulation. Oxidized forms of albumin are increased in different pathophysiological conditions and sulfenic acid lies in a mechanistic junction, relating oxidizing species to final thiol oxidation products.

L. Turell; S. Carballal; H. Botti; R. Radi; B. Alvarez

2009-01-01

232

Miniaturized capillary electrophoresis system with a carbon nanotube microelectrode for rapid separation and detection of thiols.  

Science.gov (United States)

Multi-walled carbon nanotube (CNT) was mixed with epoxy to fabricate microdisc electrode used as a detector for a specially designed miniaturized capillary electrophoresis (CE)-amperometric detection system for the separation and detection of several bioactive thiols. The end-channel CNT amperometric detector offers favourable signal-to-noise characteristics at a relatively low potential (0.8V) for detecting thiol compounds. Factors influencing the separation and detection processes were examined and optimized. Four thiols (homocysteine, cysteine, glutathione, and N-acetylcysteine) have been separated within 130s at a separation voltage of 2000V using a 20mM phosphate running buffer (pH 7.8). Highly linear response is obtained for homocysteine, cysteine, glutathione, and N-acetylcysteine over the range of 5-50muM with detection limits of 0.75, 0.8, 2.9, and 3.3muM, respectively. Good stability and reproducibility (R.S.D. < 5%) are obtained reflecting the minimal adsorption of thiols at the CNT electrode surface. The new microchip protocol should find a wide range of bioanalytical applications involving assays of thiol compounds. PMID:18969705

Chen, Gang; Zhang, Luyan; Wang, Joseph

2004-11-15

233

Miniaturized capillary electrophoresis system with a carbon nanotube microelectrode for rapid separation and detection of thiols.  

UK PubMed Central (United Kingdom)

Multi-walled carbon nanotube (CNT) was mixed with epoxy to fabricate microdisc electrode used as a detector for a specially designed miniaturized capillary electrophoresis (CE)-amperometric detection system for the separation and detection of several bioactive thiols. The end-channel CNT amperometric detector offers favourable signal-to-noise characteristics at a relatively low potential (0.8V) for detecting thiol compounds. Factors influencing the separation and detection processes were examined and optimized. Four thiols (homocysteine, cysteine, glutathione, and N-acetylcysteine) have been separated within 130s at a separation voltage of 2000V using a 20mM phosphate running buffer (pH 7.8). Highly linear response is obtained for homocysteine, cysteine, glutathione, and N-acetylcysteine over the range of 5-50muM with detection limits of 0.75, 0.8, 2.9, and 3.3muM, respectively. Good stability and reproducibility (R.S.D. < 5%) are obtained reflecting the minimal adsorption of thiols at the CNT electrode surface. The new microchip protocol should find a wide range of bioanalytical applications involving assays of thiol compounds.

Chen G; Zhang L; Wang J

2004-11-01

234

Characterization of erythrose reductases from filamentous fungi.  

UK PubMed Central (United Kingdom)

Proteins with putative erythrose reductase activity have been identified in the filamentous fungi Trichoderma reesei, Aspergillus niger, and Fusarium graminearum by in silico analysis. The proteins found in T. reesei and A. niger had earlier been characterized as glycerol dehydrogenase and aldehyde reductase, respectively. Corresponding genes from all three fungi were cloned, heterologously expressed in Escherichia coli, and purified. Subsequently, they were used to establish optimal enzyme assay conditions. All three enzymes strictly require NADPH as cofactor, whereas with NADH no activity could be observed. The enzymatic characterization of the three enzymes using ten substrates revealed high substrate specificity and activity with D-erythrose and D-threose. The enzymes from T. reesei and A. niger herein showed comparable activities, whereas the one from F. graminearum reached only about a tenth of it for all tested substrates. In order to proof in vivo the proposed enzyme function, we overexpressed the erythrose reductase-encoding gene in T. reesei. An increased production of erythritol by the recombinant strain compared to the parental strain could be detected.

Jovanovi? B; Mach RL; Mach-Aigner AR

2013-01-01

235

Characterization of erythrose reductases from filamentous fungi.  

Science.gov (United States)

Proteins with putative erythrose reductase activity have been identified in the filamentous fungi Trichoderma reesei, Aspergillus niger, and Fusarium graminearum by in silico analysis. The proteins found in T. reesei and A. niger had earlier been characterized as glycerol dehydrogenase and aldehyde reductase, respectively. Corresponding genes from all three fungi were cloned, heterologously expressed in Escherichia coli, and purified. Subsequently, they were used to establish optimal enzyme assay conditions. All three enzymes strictly require NADPH as cofactor, whereas with NADH no activity could be observed. The enzymatic characterization of the three enzymes using ten substrates revealed high substrate specificity and activity with D-erythrose and D-threose. The enzymes from T. reesei and A. niger herein showed comparable activities, whereas the one from F. graminearum reached only about a tenth of it for all tested substrates. In order to proof in vivo the proposed enzyme function, we overexpressed the erythrose reductase-encoding gene in T. reesei. An increased production of erythritol by the recombinant strain compared to the parental strain could be detected. PMID:23924507

Jovanovi?, Birgit; Mach, Robert L; Mach-Aigner, Astrid R

2013-08-08

236

Nitrate Reductase from Monoraphidium braunii: Immunocytochemical Localization and Immunological Characterization.  

Science.gov (United States)

Homogeneous nitrate reductase (EC 1.6.6.2) from Monoraphidium braunii was obtained by means of affinity chromatography in blue-Sepharose and gel filtration. After electrophoresis in polyacrylamide, gel slices containing pure nitrate reductase were disrupted and injected into previously unimmunized rabbits. The antiserum produced after several weeks was found to inhibit the different activities of nitrate reductase to a similar degree. Monospecificity of the antiserum was demonstrated by Ouchterlony double diffusion and crossed immunoelectrophoresis. The antibodies were purified by immunoabsorption to Sepharose-bound nitrate reductase.The intracellular location of nitrate reductase in green algae was examined by applying an immunocytochemical method to M. braunii cells. Ultrathin frozen sections were first treated with immunopurified anti-nitrate reductase monospecific antibodies, followed by incubation with colloidal gold-labeled goat antirabbit immunoglobulin G as a marker. The enzyme was specifically located in the pyrenoid region of the chloroplast. PMID:16664292

Lopez-Ruiz, A; Roldan, J M; Verbelen, J P; Diez, J

1985-07-01

237

Nitrate Reductase from Monoraphidium braunii: Immunocytochemical Localization and Immunological Characterization.  

UK PubMed Central (United Kingdom)

Homogeneous nitrate reductase (EC 1.6.6.2) from Monoraphidium braunii was obtained by means of affinity chromatography in blue-Sepharose and gel filtration. After electrophoresis in polyacrylamide, gel slices containing pure nitrate reductase were disrupted and injected into previously unimmunized rabbits. The antiserum produced after several weeks was found to inhibit the different activities of nitrate reductase to a similar degree. Monospecificity of the antiserum was demonstrated by Ouchterlony double diffusion and crossed immunoelectrophoresis. The antibodies were purified by immunoabsorption to Sepharose-bound nitrate reductase.The intracellular location of nitrate reductase in green algae was examined by applying an immunocytochemical method to M. braunii cells. Ultrathin frozen sections were first treated with immunopurified anti-nitrate reductase monospecific antibodies, followed by incubation with colloidal gold-labeled goat antirabbit immunoglobulin G as a marker. The enzyme was specifically located in the pyrenoid region of the chloroplast.

Lopez-Ruiz A; Roldan JM; Verbelen JP; Diez J

1985-07-01

238

Structural classification and properties of ketoacyl reductases, hydroxyacyl dehydratases and enoyl reductases.  

UK PubMed Central (United Kingdom)

Ketoacyl reductases (KRs), hydroxyacyl dehydratases (HDs) and enoyl reductases (ERs) are part of the fatty acid and polyketide synthesis cycles. Their reverse reactions, catalyzed by acyl dehydrogenases (equivalent to ERs), enoyl hydratases (equivalent to HDs) and hydroxyacyl dehydrogenases (equivalent to KRs), are part of fatty acid degradation by ?-oxidation. These enzymes have been classified into families based on similarities in their primary and tertiary structures, and these families and their structures are included in the ThYme (Thioester-active enzYmes) database. Members of each family have strong sequence similarity and have essentially the same tertiary structure, mechanism and catalytic residues.

Cantu DC; Dai T; Beversdorf ZS; Reilly PJ

2012-12-01

239

Nitrate reductase of green algae is located in the pyrenoid.  

Science.gov (United States)

Antibodies against nitrate reductase from Monoraphidium braunii have been used to determine the antigenic relationships of nitrate reductases from different green algae. Nitrate reductases from Chlamydomonas reinhardii, Chlorella fusca, Dunaliella salina, and Scenedesmus obliquus, were inhibited by, and cross-reacted with, antibodies raised against the enzyme from Monoraphidium braunii.These antibodies were also used to determine, by immunoelectron microscopy, the intracellular location of nitrate reductase in the aforementioned green algae. In all cases, the enzyme was specifically located in the pyrenoid. PMID:16664519

Lopez-Ruiz, A; Verbelen, J P; Roldan, J M; Diez, J

1985-12-01

240

A thioredoxin reductase-like protein from the thermophile, Thermus scotoductus SA-01, displaying iron reductase activity.  

UK PubMed Central (United Kingdom)

The transition metal iron is an important element for the sustenance of life--it can function either as an electron acceptor or as a donor and serves as a cofactor in many enzymes activities. The cytoplasmic NAD(P)H-dependent ferric reductase in Thermus scotoductus SA-01 shares high sequence and structural similarity to prokaryotic thioredoxin reductases. Here we report the sequence of the ferric reductase (which is typically annotated as a thioredoxin reductase-like protein) and a comparative kinetic study with the thioredoxin reductase from SA-01. Structurally, the most noteworthy difference, immediately apparent from the protein sequence, is the absence of the disulphide redox centre in the ferric reductase. This is the first report relating the attributes of such a redox protein to its ability to reduce a ferric substrate.

Bester PA; Litthauer D; Piater LA; van Heerden E

2010-01-01

 
 
 
 
241

Thiol redox state and related enzymes in sclerotium-forming filamentous phytopathogenic fungi.  

Science.gov (United States)

Thiol redox state (TRS) reduced and oxidized components form profiles characteristic of each of the four main types of differentiation in the sclerotiogenic phytopathogenic fungi: loose, terminal, lateral-chained, and lateral-simple, represented by Rhizoctonia solani, Sclerotinia sclerotiorum, Sclerotium rolfsii, and Sclerotinia minor, respectively. A common feature of these fungi is that as their undifferentiated mycelium enters the differentiated state, it is accompanied by a decrease in the low oxidative stress-associated total reduced thiols and/or by an increase of the high oxidative stress-associated total oxidized thiols either in the sclerotial mycelial substrate or in its corresponding sclerotium, indicating a relationship between TRS-related oxidative stress and sclerotial differentiation. Moreover, the four studied sclerotium types exhibit high activities of TRS-related antioxidant enzymes, indicating the existence of antioxidant protection of the hyphae of the sclerotium medulla until conditions become appropriate for sclerotium germination. PMID:18400483

Patsoukis, Nikolaos; Georgiou, D Christos

2007-11-01

242

Inversion of reactivity of thiols in Ad/sub N/ reactions of acetylenes  

Energy Technology Data Exchange (ETDEWEB)

The rate of the reaction of arenethiols with diarylpropynones in the presence of triethylamine is described by a second-order kinetic equation (first in each of the reagents). The effect of the substituent in the aromatic ring of the thiol on the reaction rate is opposite in the regions of low (/rho//sub 2/ > 0) and high (/rho//sub 2/ < 0) concentrations of the catalyst (inversion of the reactivity of the thiols). Here the sign of /rho//sub 1/ (with respect to the substrate) is not inverted. According to the quantum-chemical calculations, the inversion of the reactivity of the thiols is due to a change in the forms in which the reagent participates with variation in the concentration of the catalyst and an associated change in the ratio of the attractive and repulsive interactions between the orbitals of the reagents.

Bodrikov, I.V.; Korshunov, S.P.; Bazhan, L.I.; Statsyuk, V.E.; Korzhova, N.V.

1988-09-10

243

The maize benzoxazinone DIMBOA reacts with glutathione and other thiols to form spirocyclic adducts.  

Science.gov (United States)

Maize, wheat and other grasses synthesise large quantities of benzoxazinones and their glucosides, which act as antifeedant and allelopathic agents. These activities are probably due to the electrophilic nature of the aglycones, however, the mechanism of their action is unclear. In biological systems, glutathione (GSH) is the major electrophile-reactive compound so the reaction of the major maize benzoxazinone DIMBOA with GSH was studied. GSH reacts with DIMBOA to form eight isomeric mono-conjugates and eight isomeric di-conjugates. Through NMR studies with the model thiol 2-mercaptoethanol, these were structurally elucidated as unusual spirocycles. Similar reactivity was observed with proteins, with cysteinyl thiols being modified by DIMBOA. The thioether bonds formed were stable and not easily reduced to the parent thiol. DIMBOA can therefore readily deplete GSH levels and irreversibly inactivate enzymes with active-site cysteine residues, with clear implications for potentially toxic effects when young grasses are ingested, whether by insect pests or humans. PMID:22342783

Dixon, David P; Sellars, Jonathan D; Kenwright, Alan M; Steel, Patrick G

2012-02-17

244

The maize benzoxazinone DIMBOA reacts with glutathione and other thiols to form spirocyclic adducts.  

UK PubMed Central (United Kingdom)

Maize, wheat and other grasses synthesise large quantities of benzoxazinones and their glucosides, which act as antifeedant and allelopathic agents. These activities are probably due to the electrophilic nature of the aglycones, however, the mechanism of their action is unclear. In biological systems, glutathione (GSH) is the major electrophile-reactive compound so the reaction of the major maize benzoxazinone DIMBOA with GSH was studied. GSH reacts with DIMBOA to form eight isomeric mono-conjugates and eight isomeric di-conjugates. Through NMR studies with the model thiol 2-mercaptoethanol, these were structurally elucidated as unusual spirocycles. Similar reactivity was observed with proteins, with cysteinyl thiols being modified by DIMBOA. The thioether bonds formed were stable and not easily reduced to the parent thiol. DIMBOA can therefore readily deplete GSH levels and irreversibly inactivate enzymes with active-site cysteine residues, with clear implications for potentially toxic effects when young grasses are ingested, whether by insect pests or humans.

Dixon DP; Sellars JD; Kenwright AM; Steel PG

2012-05-01

245

New design of thiol-responsive degradable polylactide-based block copolymer micelles.  

UK PubMed Central (United Kingdom)

A new design to synthesize thiol-responsive degradable polylactide (PLA)-based micelles having a disulfide linkage in the middle of triblock copolymers is reported. They were synthesized by a new method that centers on the use of a disulfide-labeled diol as an initiator for ring-opening polymerization, followed by controlled radical polymerization. These well-controlled copolymers with monomodal and narrow molecular weight distribution (M(w) /M(n) < 1.15) self-assembled to form aqueous micellar aggregates with disulfide-containing PLA cores, which is not toxic to cells. Central disulfide linkages were cleaved in response to thiols; such thiol-triggered degradation enhanced the release of encapsulated anticancer drugs.

Cunningham A; Oh JK

2013-01-01

246

D-penicillamine and other low molecular weight thiols: review of anticancer effects and related mechanisms.  

Science.gov (United States)

Low molecular weight thiols (LMWTs) like N-acetyl cysteine, D-penicillamine, captopril, Disulfiram and Amifostine, etc. have been used as chemo-preventive agents. Recent studies have reported cell growth inhibition and cytotoxicity in several different types of cancer cells following treatment with several LMWTs. Cytotoxic and cytostatic effects of LMWTs may involve interaction of the thiol group with cellular lipids, proteins, intermediates or enzymes. Some of the mechanisms that have been proposed include a p53 mediated apoptosis, thiyl radical induced DNA damage, membrane damage through lipid peroxidation, anti-angiogenic effects induced by inhibition of matrix metalloproteinase enzymes and angiostatin generation. LMWTs are strong chelators of transition metals like copper, nickel, zinc, iron and cobalt and may cause metal co-factor depletion resulting in cytotoxicity. Oxidation of thiol group can also generate cytotoxic reactive oxygen species (ROS). PMID:23727371

Wadhwa, Saurabh; Mumper, Russell J

2013-05-29

247

D-penicillamine and other low molecular weight thiols: review of anticancer effects and related mechanisms.  

UK PubMed Central (United Kingdom)

Low molecular weight thiols (LMWTs) like N-acetyl cysteine, D-penicillamine, captopril, Disulfiram and Amifostine, etc. have been used as chemo-preventive agents. Recent studies have reported cell growth inhibition and cytotoxicity in several different types of cancer cells following treatment with several LMWTs. Cytotoxic and cytostatic effects of LMWTs may involve interaction of the thiol group with cellular lipids, proteins, intermediates or enzymes. Some of the mechanisms that have been proposed include a p53 mediated apoptosis, thiyl radical induced DNA damage, membrane damage through lipid peroxidation, anti-angiogenic effects induced by inhibition of matrix metalloproteinase enzymes and angiostatin generation. LMWTs are strong chelators of transition metals like copper, nickel, zinc, iron and cobalt and may cause metal co-factor depletion resulting in cytotoxicity. Oxidation of thiol group can also generate cytotoxic reactive oxygen species (ROS).

Wadhwa S; Mumper RJ

2013-08-01

248

Aqueous Synthesis of Peptide Thioesters from Amino Acids and a Thiol Using 1,1?-Carbonyldiimidazole  

Science.gov (United States)

A new method was developed for the synthesis of peptide thioesters from free amino acids and thiols in water. This one-pot simple method involves two steps: (1) activation in water of an amino acid presumably as its N-carboxyanhydride (NCA) using 1,1?-carbonyldiimidazole (CDI), and (2) subsequent condensation of the activated amino acid-NCA in the presence of a thiol. With this method citrulline peptide thioesters containing up to 10 amino acid residues were prepared in a single reaction. This aqueous synthetic method provides a simple way to prepare peptide thioesters for studies of peptide replication involving ligation of peptide thioesters on peptide templates. The relevance of peptide replication to the origin-of-life process is supported by previous studies showing that amino acid thioesters (peptide thioester precursors) can be synthesized under prebiotic conditions by reaction of small sugars with ammonia and a thiol.

Weber, Arthur L.

2005-10-01

249

Influence of volatile thiols in the development of blackcurrant aroma in red wine.  

UK PubMed Central (United Kingdom)

A strong blackcurrant aroma was recently perceived in some red wines originating from the same appellation. Varietal thiols such as 4-mercapto-4-methyl-2-pentanone (4MMP), 3-(mercapto)hexyl acetate (3MHA) and 3-mercapto-1-hexanol (3MH) are compounds potentially responsible for the development of this aroma. In order to demonstrate the correlation between thiols concentrations in red wines and blackcurrant aroma intensity, a multiple variable analysis was realised with thiols concentrations obtained by chemical analysis and blackcurrant aroma intensities obtained by descriptive sensory analysis. The 4MMP concentration was very well correlated to the blackcurrant aroma, and 3MHA and 3MH present at high concentrations act as enhancers of the perception of this aroma. This correlation was further supported after performing a sensory comparison by classification test. The different factors that could impact on the development of blackcurrant aroma in red wine were discussed.

Rigou P; Triay A; Razungles A

2014-01-01

250

Influence of volatile thiols in the development of blackcurrant aroma in red wine.  

Science.gov (United States)

A strong blackcurrant aroma was recently perceived in some red wines originating from the same appellation. Varietal thiols such as 4-mercapto-4-methyl-2-pentanone (4MMP), 3-(mercapto)hexyl acetate (3MHA) and 3-mercapto-1-hexanol (3MH) are compounds potentially responsible for the development of this aroma. In order to demonstrate the correlation between thiols concentrations in red wines and blackcurrant aroma intensity, a multiple variable analysis was realised with thiols concentrations obtained by chemical analysis and blackcurrant aroma intensities obtained by descriptive sensory analysis. The 4MMP concentration was very well correlated to the blackcurrant aroma, and 3MHA and 3MH present at high concentrations act as enhancers of the perception of this aroma. This correlation was further supported after performing a sensory comparison by classification test. The different factors that could impact on the development of blackcurrant aroma in red wine were discussed. PMID:24001837

Rigou, Peggy; Triay, Aurélie; Razungles, Alain

2013-07-17

251

Hybrid thiol-ene network nanocomposites based on multi(meth)acrylate POSS.  

UK PubMed Central (United Kingdom)

First, multi(meth)acrylate functionalized POSS monomers were synthesized in this paper. Secondly, FTIR was used to evaluate the homopolymerization behaviors of multi(meth)acrylate POSS and their copolymerization behaviors in the thiol-ene reactions with octa(3-mercaptopropyl) POSS in the presence of photoinitiator. Results showed that the photopolymerization rate of multimethacrylate POSS was faster than that of multiacrylate POSS. The FTIR results also showed that the copolymerizations were dominant in the thiol-ene reactions with octa(3-mercaptopropyl) POSS, different from traditional (meth)acrylate-thiol system, in which homopolymerizations were predominant. Finally, the resulted hybrid networks based on POSS were characterized by XRD, FE-SEM, DSC, and TGA. The characterization results showed that hybrid networks based on POSS were homogeneous and exhibited high thermal stability.

Li L; Liang R; Li Y; Liu H; Feng S

2013-09-01

252

Engineering volatile thiol release in Saccharomyces cerevisiae for improved wine aroma.  

Science.gov (United States)

Volatile thiols, such as 4-mercapto-4-methylpentan-2-one (4MMP), 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl acetate (3MHA), are among the most potent aroma compounds found in wine and can have a significant effect on wine quality and consumer preferences. At optimal concentrations in wine, these compounds impart flavours of passionfruit, grapefruit, gooseberry, blackcurrant, lychee, guava and box hedge. The enzymatic release of aromatic thiols from grape-derived, non-volatile cysteinylated precursors (Cys-4MMP and Cys-3MH) and the further modification thereof (conversion of 3MH into 3MHA) during fermentation, enhance the varietal characters of wines such as Sauvignon Blanc. Wine yeast strains have limited and varying capacities to produce aroma-enhancing thiols from their non-volatile counterparts in grape juice. Even under optimal fermentation conditions, the most efficient thiol-releasing Saccharomyces cerevisiae wine strain known realizes less than 5% of the thiol-related flavour potential of grape juice. The objective of this study was to develop a wine yeast able to unleash the untapped thiol aromas in grape juice during winemaking. To achieve this goal, the Escherichia coli tnaA gene, encoding a tryptophanase with strong cysteine-beta-lyase activity, was cloned and overexpressed in a commercial wine yeast strain under the control of the regulatory sequences of the yeast phosphoglycerate kinase I gene (PGK1). This modified strain expressing carbon-sulphur lyase activity released up to 25 times more 4MMP and 3MH in model ferments than the control host strain. Wines produced with the engineered strain displayed an intense passionfruit aroma. This yeast offers the potential to enhance the varietal aromas of wines to predetermined market specifications. PMID:17492802

Swiegers, Jan H; Capone, Dimitra L; Pardon, Kevin H; Elsey, Gordon M; Sefton, Mark A; Francis, I Leigh; Pretorius, Isak S

2007-07-01

253

Production of Thiol Species From An Exponential Growth Diatom Under Copper Exposure  

Science.gov (United States)

The intracellar induction of phytochelatins is a well documented response of eukaryotic microorganisms to aqueous metal exposure. The extracellular release of thiolic compounds from algal species has been observed; and in some cases, this release can contribute a significant fraction of the uncharacterized metal-complexing ligands. Glutathione (GSH) or cysteine is among the detectable thiols excreted. A quantitative assessment of the excretion of thiols from algae cells into growth media is needed to assess the significance of biogenic-thiols as a source of strong ligands in natural waters and as a "forgotten" route in sulfur biogeochemical cycle. Unbuffered growth media (e.g., without adding complexing ligand such as EDTA) have only rarely been used to study the possible release of metal-complexing ligands from algal species, and the ligand titration techniques used varied considerably. The majority of culture studies have applied metal-buffered media. A direct comparison of released ligands under buffered and unbuffered conditions is lacking, partially due to the inherent difficulties of the titration methods applied. Using HPLC with fluorescence detection of thiol-monobromobimane derivatives, we were able to follow the dynamic change of GSH released in both media types during algal growth: (1) the cell quotas for thiols and pigments varied (mostly decreases) with growth time. Therefore, pigment-normalized cellular thiol concentrations were more or less conservative. (2) GSH was released into both the EDTA-buffered and -unbuffered growth media at similar concentrations. (3) at similar available Cu concentrations, EDTA possibly enhanced, rather than hindered, the release of GSH.

Tang, D.; Shafer, M. M.; Karner, D. A.; Armstrong, D. E.; Schauer, J.

2003-12-01

254

Protein thiol oxidation in murine airway epithelial cells in response to naphthalene or diethyl maleate.  

UK PubMed Central (United Kingdom)

Naphthalene (NA) is a semivolatile aromatic hydrocarbon to which humans are exposed from a variety of sources. NA results in acute cytotoxicity to respiratory epithelium in rodents. Cytochrome P450-dependent metabolic activation to form reactive intermediates and loss of soluble cellular thiols (glutathione) are critical steps in NA toxicity, but the precise mechanisms by which this chemical results in cellular injury remain unclear. Protein thiols are likely targets of reactive NA metabolites. Loss of these, through adduction or thiol oxidation mechanisms, may be important underlying mechanisms for NA toxicity. To address the hypothesis that loss of thiols on specific cellular proteins is critical to NA-induced cytotoxicity, we compared reduced to oxidized thiol ratios in airway epithelial cell proteins isolated from lungs of mice treated with NA or the nontoxic glutathione depletor, diethyl maleate (DEM). At 300 mg/kg doses, NA administration resulted in a greater than 85% loss of glutathione levels in the airway epithelium, which is similar to the loss observed after DEM treatment. Using differential fluorescent maleimide labeling followed by 2DE separation of proteins, we identified more than 35 unique proteins that have treatment-specific differential sulfhydryl oxidation. At doses of NA and DEM that produce similar levels of glutathione depletion, Cy3/Cy5 labeling ratios were statistically different for 16 nonredundant proteins in airway epithelium. Proteins identified include a zinc finger protein, several aldehyde dehydrogenase variants, beta-actin, and several other structural proteins. These studies show distinct patterns of protein thiol alterations with the noncytotoxic DEM and the cytotoxic NA.

Spiess PC; Morin D; Williams CR; Buckpitt AR

2010-09-01

255

Visible-light-mediated thiol-ene hydrogelation using eosin-Y as the only photoinitiator.  

UK PubMed Central (United Kingdom)

The utility of visible-light-mediated polymerization in tissue engineering has been limited due to the necessary use of potentially cytotoxic coinitiator and comonomer. Here, we report a visible-light-mediated thiol-ene hydrogelation scheme using eosin-Y as the only photoinitiator. Under visible light exposure, rapid and highly tunable step-growth gelation is achieved using PEG-norbornene and a model cross-linker dithiothreitol. In addition to investigating the gelation kinetics and properties of thiol-ene hydrogels formed by this new gelation scheme, we also report high cytocompatibility of these hydrogels using human mesenchymal stem cells (hMSCs) and pancreatic MIN6 ?-cells.

Shih H; Lin CC

2013-02-01

256

Self-assembled monolayers of semifluorinated thiols on electrochemically modified polycrystalline nickel surfaces  

Energy Technology Data Exchange (ETDEWEB)

Electrochemically pretreated polycrystalline nickel substrates modified with ethanolic solutions (10{sup -2} and 10{sup -1} M) of four semi-fluorinated thiols with R {sub F}-R {sub H}-SH structures have been evaluated by X-ray photoelectron spectroscopy, contact angles and cyclic voltammetry measurements. Our results show that it is possible to graft highly fluorinated alkanethiols on nickel surfaces, the quality of the self assembled monolayers depending on the concentration of the dipping solutions, the dipping time as well as the length of the perfluorinated carbon chains and those of the hydrocarbon connector between the perfluorinated fragment and the thiol function.

Tortech, L. [Laboratoire de Chimie des Materiaux Organiques et Metalliques, CMOM, Faculte des Sciences, Universite de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2 (France); Laboratoire Interdisciplinaire de Spectroscopie Electronique, Facultes Universitaires Notre-Dame de la Paix, rue de Bruxelles, 61, B-5000 Namur (Belgium); Mekhalif, Z. [Laboratoire Interdisciplinaire de Spectroscopie Electronique, Facultes Universitaires Notre-Dame de la Paix, rue de Bruxelles, 61, B-5000 Namur (Belgium); Delhalle, J. [Laboratoire Interdisciplinaire de Spectroscopie Electronique, Facultes Universitaires Notre-Dame de la Paix, rue de Bruxelles, 61, B-5000 Namur (Belgium); Guittard, F. [Laboratoire de Chimie des Materiaux Organiques et Metalliques, CMOM, Faculte des Sciences, Universite de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2 (France); Geribaldi, S. [Laboratoire de Chimie des Materiaux Organiques et Metalliques, CMOM, Faculte des Sciences, Universite de Nice-Sophia Antipolis, Parc Valrose, 06108 Nice Cedex 2 (France)]. E-mail: geribald@unice.fr

2005-11-22

257

Self-assembled monolayers of semifluorinated thiols on electrochemically modified polycrystalline nickel surfaces  

International Nuclear Information System (INIS)

Electrochemically pretreated polycrystalline nickel substrates modified with ethanolic solutions (10-2 and 10-1 M) of four semi-fluorinated thiols with R F-R H-SH structures have been evaluated by X-ray photoelectron spectroscopy, contact angles and cyclic voltammetry measurements. Our results show that it is possible to graft highly fluorinated alkanethiols on nickel surfaces, the quality of the self assembled monolayers depending on the concentration of the dipping solutions, the dipping time as well as the length of the perfluorinated carbon chains and those of the hydrocarbon connector between the perfluorinated fragment and the thiol function.

2005-11-22

258

Synthesis of Novel Hexathiolated Squalene and Its Thiol-Ene Photopolymerization with Unsaturated Monomers  

Directory of Open Access Journals (Sweden)

Full Text Available In this work is described the synthesis of a multifunctional thiolated squalene. Thiol-ene coupling reactions were employed to functionalize the six double bonds of squalene, using thiolacetic acid. Hydrolysis of the resulting thioacetates, rendered the corresponding hexathiolated squalene SQ6SH. This compound was further photopolymerized separately with triallyl cyanurate, pentaerythritol triacrylate and diethyleneglycol divinyl ether. Real Time FTIR kinetics revealed that homopolymerization of the ene monomers took place in addition to the thiol-ene photopolymerization. Flexible films were obtained when SQ6SH was photopolymerized in bulk with the above mentioned unsaturated monomers.

Ricardo Acosta Ortiz; Ena Adeligna Obregón Blandón; Ramiro Guerrero Santos

2012-01-01

259

High-performance liquid chromatographic separation and indirect fluorescence detection of thiols.  

UK PubMed Central (United Kingdom)

A fluorescent post-column reaction detection scheme has been devised for selective determination of thiols. The post-column reagent is 40 microM Cd2+ and 100 microM 8-hydroxyquinoline-5-sulfonic acid (HQS) in non-complexing buffer at pH 10. HQS complexes Cd2+ to form a fluorescent product. Thiols in the HPLC effluent compete for complexation of the Cd2+, resulting in a decrease in the fluorescence response. Detection limits of 0.2 microM (0.04 ppm) are achieved for cysteine, homocysteine and glutathione in a 5 min separation. Recoveries from spiked synthetic urine samples are 87-120%.

Pelletier S; Lucy CA

2002-10-01

260

Mechanisms and dynamics in the thiol/disulfide redox regulatory network: transmitters, sensors and targets.  

UK PubMed Central (United Kingdom)

Plant cells sense, weigh and integrate various endogenous and exogenous cues in order to optimize acclimation and resource allocation. The thiol/disulfide redox network appears to be in the core of this versatile integration process. In plant cells its complexity exceeds by far that of other organisms. Recent research has elucidated the multiplicity of the diversified input elements, transmitters (thioredoxin, glutaredoxins), targets and sensors (peroxiredoxins and other peroxidases), controlled processes and final acceptors (reactive oxygen species). An additional level of thiol/disulfide regulation is achieved by introducing dynamics in time and subcompartment and complex association.

König J; Muthuramalingam M; Dietz KJ

2012-06-01

 
 
 
 
261

NMR study of 2-amino-cyclo-hexane thiols N-substituted  

International Nuclear Information System (INIS)

Considering that thiols and mainly ?-aminoacids are been successful in research aiming the discovering of new substances with protective actions against the radiation effects on living being, 2-amine-cyclo-hexane thiols N-alkyl-substituted were synthesized and submitted to to biological tests. The reactions were planned to favor the SN 2 mechanism, in order to obtain the respective trans isomers from each ?-aminothiol (nonpolar medium; solvent: benzene or hexane). This work presents the results obtained in the NMR study of four by products of this class

1999-01-01

262

A first principles study of thiol-capped Au nanoparticles: structural, electronic, and magnetic properties as a function of thiol coverage.  

UK PubMed Central (United Kingdom)

We have studied the stability of thiolated Au38 nanoparticles (NPs) via density functional theory based calculations varying the coverage from 0 up to 32 molecules. Three different initial core arrangements were considered for the cluster, spherical, tubular, and bi-icosahedral, while thiol groups were attached to the cluster via the sulfur atom either as single molecules or forming more complex staple motifs. After molecular dynamics runs several metastable configurations are found at each coverage thus allowing to analyze the properties of the NPs in the form of ensemble averages. In particular, we address the structural and electronic properties as a function of the number of thiols. The study emphasizes the strong influence of the core structure on the stability of the NPs, and its interplay with the thiol coverage and adsorption geometries. The magnetic properties of the NPs have also been explored via spin-polarized calculations including spin-orbit coupling. No evidence for the existence of a robust intrinsic ferromagnetism is found in any of the structures.

Cuadrado R; Puerta JM; Soria F; Cerdá JI

2013-07-01

263

[Tetrathionate reductase as a diagnostic trait. I. Medium for tetrathionate reductase determination  

UK PubMed Central (United Kingdom)

In a medium proposed for the determination of tetrathionate reductase potassium tetrathionate, requiring complicated techniques for preparation and decomposing during sterilization, is replaced by sodium tetrathionate, obtained extemporaneously and simple in preparation. The medium is balanced and contains no free iodine and sodium thiosulfate.

Kalina GP; Trukhina GM

1983-02-01

264

Structure and function of NADPH-cytochrome P450 reductase and nitric oxide synthase reductase domain  

International Nuclear Information System (INIS)

NADPH-cytochrome P450 reductase (CPR) and the nitric oxide synthase (NOS) reductase domains are members of the FAD-FMN family of proteins. The FAD accepts two reducing equivalents from NADPH (dehydrogenase flavin) and FMN acts as a one-electron carrier (flavodoxin-type flavin) for the transfer from NADPH to the heme protein, in which the FMNH ?/FMNH2 couple donates electrons to cytochrome P450 at constant oxidation-reduction potential. Although the interflavin electron transfer between FAD and FMN is not strictly regulated in CPR, electron transfer is activated in neuronal NOS reductase domain upon binding calmodulin (CaM), in which the CaM-bound activated form can function by a similar mechanism to that of CPR. The oxygenated form and spin state of substrate-bound cytochrome P450 in perfused rat liver are also discussed in terms of stepwise one-electron transfer from CPR. This review provides a historical perspective of the microsomal mixed-function oxidases including CPR and P450. In addition, a new model for the redox-linked conformational changes during the catalytic cycle for both CPR and NOS reductase domain is also discussed.

2005-12-09

265

Colorimetric and luminescent dual-signaling responsive probing of thiols by a ruthenium(II)-azo complex.  

UK PubMed Central (United Kingdom)

A dinuclear ruthenium(II) complex linked via a reducible azo group [Ru(bpy)2(azobpy)Ru(bpy)2]Cl4 (Ru2azo, bpy=2,2'-bipyridine, azobpy=4,4?-azobis (2,2'-bipyridine)) was adopted as a probe for thiols. Results showed that Ru2azo could selectively and effectively react with biological thiols (such as cysteine, homocysteine and glutathione) with a 10(-7)M detection limit. After it reacted with thiols, the original gray color of Ru2azo solution immediately turned yellow and the luminescence significantly enhanced, showing "naked-eye" colorimetric and "off-on" luminescent dual-signaling response for thiols. Mechanism studies demonstrated that Ru2azo reacted with thiols undergoing a two-electron transfer process, forming the azo(2-) anion product.

Li GY; Liu JP; Huang HY; Wen Y; Chao H; Ji LN

2013-04-01

266

Colorimetric and luminescent dual-signaling responsive probing of thiols by a ruthenium(II)-azo complex.  

Science.gov (United States)

A dinuclear ruthenium(II) complex linked via a reducible azo group [Ru(bpy)2(azobpy)Ru(bpy)2]Cl4 (Ru2azo, bpy=2,2'-bipyridine, azobpy=4,4?-azobis (2,2'-bipyridine)) was adopted as a probe for thiols. Results showed that Ru2azo could selectively and effectively react with biological thiols (such as cysteine, homocysteine and glutathione) with a 10(-7)M detection limit. After it reacted with thiols, the original gray color of Ru2azo solution immediately turned yellow and the luminescence significantly enhanced, showing "naked-eye" colorimetric and "off-on" luminescent dual-signaling response for thiols. Mechanism studies demonstrated that Ru2azo reacted with thiols undergoing a two-electron transfer process, forming the azo(2-) anion product. PMID:23376332

Li, Guan-Ying; Liu, Jiang-Ping; Huang, Huai-Yi; Wen, Ya; Chao, Hui; Ji, Liang-Nian

2013-01-12

267

Preparation of a novel carboxyl stationary phase by "thiol-ene" click chemistry for hydrophilic interaction chromatography.  

UK PubMed Central (United Kingdom)

A novel carboxyl-bonded silica stationary phase was prepared by "thiol-ene" click chemistry. The resultant Thiol-Click-COOH phase was evaluated under hydrophilic interaction liquid chromatography (HILIC) mobile phase conditions. A comparison of the chromatographic performance of Thiol-Click-COOH and pure silica columns was performed according to the retention behaviors of analytes and the charged state of the stationary phases. The results indicated that the newly developed Thiol-Click-COOH column has a higher surface charge and stronger hydrophilicity than the pure silica column. Furthermore, the chromatographic behaviors of five nucleosides on the Thiol-Click-COOH phase were investigated in detail. Finally, a good separation of 13 nucleosides and bases, and four water-soluble vitamins was achieved.

Peng XT; Liu T; Ji SX; Feng YQ

2013-08-01

268

Simultaneous detection of N-acetyl-L-cysteine and physiological low molecular mass thiols in plasma by capillary electrophoresis.  

Science.gov (United States)

N-acetyl-L-cysteine (NAC) is a therapeutic drug widely used as mucolytic agent in the treatment of respiratory diseases. Recently it has been proposed that NAC administration may modify the plasma levels of low molecular weight thiols (LMW) like cysteine, homocysteine and glutathione, though it has been still debated if their plasma concentration increases or decreases during the therapy. Therefore research calls for methods able to analyze simultaneously NAC and the other plasma LMW thiols in order to evaluate if NAC is able to modify plasma thiols concentration and in particular to reduce homocysteine levels in hyperhomocysteinemia. In this paper we present a new capillary electrophoresis method that allows a baseline separation of plasma NAC from the physiological thiols. The proposed method has been utilized to measure the drug and the physiological LMW thiols in NAC administered chronic obstructive broncho-pneumopathy (COPB) disease patients. PMID:18695935

Zinellu, Angelo; Sotgia, Salvatore; Scanu, Bastianina; Usai, Maria Franca; Fois, Alessandro Giuseppe; Spada, Valentina; Deledda, Anna; Deiana, Luca; Pirina, Piero; Carru, Ciriaco

2008-08-11

269

On the mechanism and rate of gold incorporation into thiol-dependent flavoreductases.  

UK PubMed Central (United Kingdom)

NADPH-dependent flavoreductases are important drug targets. During their enzymatic cycle thiolates and selenolates that have high affinity for transition metals are generated. Auranofin (AF), a gold-containing compound, is classified by the World Health Organization as an antirheumatic agent and it is indicated as the scaffold for the development of new anticancer and antiparasitic drugs. AF inhibits selenocysteine-containing flavoreductases (thioredoxin reductase and thioredoxin glutathione reductase) more effectively than non Se-containing ones (glutathione reductase); this preference has been ascribed to the high affinity of selenium for gold. We solved the 3D structure of the Se-containing Thioredoxin Glutathione Reductase from the human parasite Schistosoma mansoni complexed with Au and our results challenge this view: we believe that the relative velocity of the reaction rather than the relative affinity, depends on the presence of Sec residues, which appear to dictate AF selectivity.

Saccoccia F; Angelucci F; Boumis G; Brunori M; Miele AE; Williams DL; Bellelli A

2012-03-01

270

5 ?-Reductase Type 2 Deficiency: Response to Dihydrotestosterone Gel.  

Science.gov (United States)

5?-reductase (5?-R) deficiency is an important cause of ambiguious genitalia in genetic males; however therapeutic experience in literature is limited. In this report authors describe a child with 46 XY Disorder of Sexual Differentiation (DSD), due to 5?-reductase deficiency, who was managed with Dihydrotestosterone (DHT) gel. PMID:23702801

Vupputuri, Madhavarao; Kandepu, Madhurima; Devireddy, Harikishore Reddy

2013-05-24

271

5 ?-Reductase Type 2 Deficiency: Response to Dihydrotestosterone Gel.  

UK PubMed Central (United Kingdom)

5?-reductase (5?-R) deficiency is an important cause of ambiguious genitalia in genetic males; however therapeutic experience in literature is limited. In this report authors describe a child with 46 XY Disorder of Sexual Differentiation (DSD), due to 5?-reductase deficiency, who was managed with Dihydrotestosterone (DHT) gel.

Vupputuri M; Kandepu M; Devireddy HR

2013-05-01

272

Dynamic Control of Electron Transfers in Diflavin Reductases  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Diflavin reductases are essential proteins capable of splitting the two-electron flux from reduced pyridine nucleotides to a variety of one electron acceptors. The primary sequence of diflavin reductases shows a conserved domain organization harboring two catalytic domains bound to the FAD and FMN f...

Louise Aigrain; Fataneh Fatemi; Oriane Frances; Ewen Lescop; Gilles Truan

273

Molecular modeling of the reductase domain to elucidate the reaction mechanism of reduction of peptidyl thioester into its corresponding alcohol in non-ribosomal peptide synthetases  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract Background Nonribosomal peptide synthetases (NRPSs) are multienzymatic, multidomain megasynthases involved in the biosynthesis of pharmaceutically important nonribosomal peptides. The peptaibol synthetase from Trichoderma virens (TPS) is an important member of the NRPS family that exhibits antifungal properties. The majority of the NRPSs terminate peptide synthesis with the thioesterase (TE) domain, which either hydrolyzes the thioester linkage, releasing the free peptic acid, or catalyzes the intramolecular macrocyclization to produce a macrolactone product. TPS is an important NRPS that does not encompass a TE domain, but rather a reductase domain (R domain) to release the mature peptide product reductively with the aid of a NADPH cofactor. However, the catalytic mechanism of the reductase domain has not yet been elucidated. Results We present here a three-dimensional (3D) model of the reductase domain based on the crystal structure of vestitone reductase (VR). VR belongs to the short-chain dehydrogenase/reductase (SDR) superfamily and is responsible for the nicotinamide dinucleotide phosphate (NADPH)-dependent reduction of the substrate into its corresponding secondary alcohol product. The binding sites of the probable linear substrates, alamethicin, trichotoxin, antiamoebin I, chrysopermin C and gramicidin, were identified within the modeled R domain using multiple docking approaches. The docking results of the ligand in the active site of the R domain showed that reductase side chains have a high affinity towards ligand binding, while the thioester oxygen of each substrate forms a hydrogen bond with the OH group of Tyr176 and the thiol group of the substrate is closer to the Glu220. The modeling and docking studies revealed the reaction mechanism of reduction of thioester into a primary alcohol. Conclusion Peptaibol biosynthesis incorporates a single R domain, which appears to catalyze the four-electron reduction reaction of a peptidyl carrier protein (PCP)-bound peptide to its corresponding primary alcohol. Analysis of R domains present in the non-redundant (nr) database of the NCBI showed that the R domain always resides in the last NRPS module and is involved in either a two or four-electron reduction reaction.

Manavalan Balachandran; Murugapiran Senthil K; Lee Gwang; Choi Sangdun

2010-01-01

274

Crystal Growth of Thiol-Stabilized Gold Nanoparticles by Heat-Induced Coalescence  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A monolayer of dodecanethiol-stabilized gold nanoparticles changed into two-dimensional and three-dimensional self-organized structures by annealing at 323 K. Subsequent crystal growth of gold nanoparticles occurred. Thiol molecules, although chemisorbed, form relatively unstable bonds with the gold...

Moon, Sook Young; Tanaka, Shun-ichiro; Sekino, Tohru

275

Optimization of Optical Properties of Polycarbonate Film with Thiol Gold-Nanoparticles  

Digital Repository Infrastructure Vision for European Research (DRIVER)

A new nanostructured composite film based on thiol gold nanoparticles dispersed in polycarbonate and prepared by evaporating a solution of 1-dodecanthiol gold nanoparticles and polycarbonate was developed for applications as optical lenses. Lenses with superior mechanical properties, coloring and UV...

Claudio Larosa; Enrico Stura; Roberto Eggenhöffner; Claudio Nicolini

276

Disulfide/thiol switches in thiosemicarbazone ligands for redox-directed iron chelation.  

UK PubMed Central (United Kingdom)

A disulfide bond is incorporated in the scaffold of thiosemicarbazone iron chelators as a reduction/activation switch. Following reduction, thiol-containing ligands stabilize iron ions in their trivalent oxidation state. The antiproliferative activity of the new chelating systems is assessed in human cancer cell lines and in normal tissue.

Chang TM; Tomat E

2013-06-01

277

Factors influencing the oxidation of cysteamine and other thiols: implications for hyperthermic sensitization and radiation protection  

International Nuclear Information System (INIS)

Some of the factors influencing the oxygen uptake and peroxide formation for cysteamine (MEA) and other thiols in serum-supplemented modified McCoy's 5A, a well-known medium used to cultivate a variety of cells in vitro, have been studied. The oxidation of MEA and cysteine in modified McCoy's 5A has been compared with that in Ham's F-12, MEM, and phosphate-buffered saline. The ability to produce peroxide is dependent upon the temperature, the concentration of thiol, the presence of copper ions, and pH of the medium. Catalase also reduces the oxygen uptake for all thiols. Superoxide dismutase (SOD) was found to stimulate the oxygen uptake in the case of MEA and cysteine, but had little or no effect with DTT and glutathione. The combined presence of SOD and catalase resulted in less inhibition of oxygen uptake than that obtained by catalase alone. Alkaline pH was found to enhance the oxidation of cysteine and MEA. The results indicate that many problems may arise when thiols are added to various media. A major consideration is concerned with the production of peroxide, superoxide, and reduced trace metal intermediates. The presence of these intermediates may result in the production of hydroxyl radical intermediates as well as the eventual oxygen depletion from the medium.

1984-01-01

278

Thimerosal Exposure and the Role of Sulfation Chemistry and Thiol Availability in Autism  

Directory of Open Access Journals (Sweden)

Full Text Available Autism spectrum disorder (ASD) is a neurological disorder in which a significant number of the children experience a developmental regression characterized by a loss of previously acquired skills and abilities. Typically reported are losses of verbal, nonverbal, and social abilities. Several recent studies suggest that children diagnosed with an ASD have abnormal sulfation chemistry, limited thiol availability, and decreased glutathione (GSH) reserve capacity, resulting in a compromised oxidation/reduction (redox) and detoxification capacity. Research indicates that the availability of thiols, particularly GSH, can influence the effects of thimerosal (TM) and other mercury (Hg) compounds. TM is an organomercurial compound (49.55% Hg by weight) that has been, and continues to be, used as a preservative in many childhood vaccines, particularly in developing countries. Thiol-modulating mechanisms affecting the cytotoxicity of TM have been identified. Importantly, the emergence of ASD symptoms post-6 months of age temporally follows the administration of many childhood vaccines. The purpose of the present critical review is provide mechanistic insight regarding how limited thiol availability, abnormal sulfation chemistry, and decreased GSH reserve capacity in children with an ASD could make them more susceptible to the toxic effects of TM routinely administered as part of mandated childhood immunization schedules.

Janet K. Kern; Boyd E. Haley; David A. Geier; Lisa K. Sykes; Paul G. King; Mark R. Geier

2013-01-01

279

Enantiospecific synthesis of [2.2]paracyclophane-4-thiol and derivatives  

Directory of Open Access Journals (Sweden)

Full Text Available This paper describes a simple route to enantiomerically enriched [2.2]paracyclophane-4-thiol via the stereospecific introduction of a chiral sulfoxide to the [2.2]paracyclophane skeleton. The first synthesis of an enantiomerically enriched planar chiral benzothiazole is also reported.

Gareth J. Rowlands; Richard J. Seacome

2009-01-01

280

Analytical utility of 2-halopyridinium salts-II Acidimetric determination of thiol groups.  

UK PubMed Central (United Kingdom)

2-Chloro-1-methylpyridinium iodide reacts rapidly and quantitatively with thiol groups in the presence of excess of triethylamine to give the 2-alkyl(aryl)thio-1-methylpyridinium iodide and an equimolar amount of hydrogen chloride which is trapped by the triethylamine. The excess of triethylamine is back-titrated with hydrochloric acid, with Bromothymol Blue as indicator.

Bald E

1980-03-01

 
 
 
 
281

Oxidative stress and pathology in muscular dystrophies: focus on protein thiol oxidation and dysferlinopathies.  

UK PubMed Central (United Kingdom)

The muscular dystrophies comprise more than 30 clinical disorders that are characterized by progressive skeletal muscle wasting and degeneration. Although the genetic basis for many of these disorders has been identified, the exact mechanism for pathogenesis generally remains unknown. It is considered that disturbed levels of reactive oxygen species (ROS) contribute to the pathology of many muscular dystrophies. Reactive oxygen species and oxidative stress may cause cellular damage by directly and irreversibly damaging macromolecules such as proteins, membrane lipids and DNA; another major cellular consequence of reactive oxygen species is the reversible modification of protein thiol side chains that may affect many aspects of molecular function. Irreversible oxidative damage of protein and lipids has been widely studied in Duchenne muscular dystrophy, and we have recently identified increased protein thiol oxidation in dystrophic muscles of the mdx mouse model for Duchenne muscular dystrophy. This review evaluates the role of elevated oxidative stress in Duchenne muscular dystrophy and other forms of muscular dystrophies, and presents new data that show significantly increased protein thiol oxidation and high levels of lipofuscin (a measure of cumulative oxidative damage) in dysferlin-deficient muscles of A/J mice at various ages. The significance of this elevated oxidative stress and high levels of reversible thiol oxidation, but minimal myofibre necrosis, is discussed in the context of the disease mechanism for dysferlinopathies, and compared with the situation for dystrophin-deficient mdx mice.

Terrill JR; Radley-Crabb HG; Iwasaki T; Lemckert FA; Arthur PG; Grounds MD

2013-09-01

282

Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations.  

UK PubMed Central (United Kingdom)

The detection of thiols on extraterrestrial bodies could provide evidence for life, as well as a host of potential prebiological or abiological processes. Here, we report a novel protocol to analyze organic thiols by microchip CE with LIF detection. Thiols were labeled with Pacific Blue C5 maleimide and analyzed by MEKC. The separation buffer consisted of 15 mM tetraborate pH 9.2 and 25 mM SDS. The optimized method provided LODs ranging from 1.4 to 15 nM. The method was validated using samples collected from geothermal pools at Hot Creek Gorge, California, which were found to contain 2-propanethiol and 1-butanethiol in the nanomolar concentration range. These samples serve as chemical analogues to material potentially present in the reducing environment of primitive Earth and also at sulfurous regions of Mars. Hence, the protocol developed here enables highly sensitive thiol analysis in samples with complexity comparable to that expected in astrobiologically relevant extraterrestrial settings. This new protocol could be readily added to the existing suite of microfluidic chemical analyses developed for in situ planetary exploration; all that is required is the incorporation of two new reagents to the payload of an existing instrument concept.

Mora MF; Stockton AM; Willis PA

2013-01-01

283

Nanoparticles under the light: click functionalization by photochemical thiol-yne reaction, towards double click functionalization.  

UK PubMed Central (United Kingdom)

A light click away: The first application of the thiol-yne reaction to nanoparticle functionalization is described (see figure). This metal-free click chemistry approach is compatible with the addition of various molecules at the surface and can be combined with CuAAC methodology to perform chemoselective double functionalization.

Demay-Drouhard P; Nehlig E; Hardouin J; Motte L; Guénin E

2013-06-01

284

Analysis of thiols by microchip capillary electrophoresis for in situ planetary investigations.  

Science.gov (United States)

The detection of thiols on extraterrestrial bodies could provide evidence for life, as well as a host of potential prebiological or abiological processes. Here, we report a novel protocol to analyze organic thiols by microchip CE with LIF detection. Thiols were labeled with Pacific Blue C5 maleimide and analyzed by MEKC. The separation buffer consisted of 15 mM tetraborate pH 9.2 and 25 mM SDS. The optimized method provided LODs ranging from 1.4 to 15 nM. The method was validated using samples collected from geothermal pools at Hot Creek Gorge, California, which were found to contain 2-propanethiol and 1-butanethiol in the nanomolar concentration range. These samples serve as chemical analogues to material potentially present in the reducing environment of primitive Earth and also at sulfurous regions of Mars. Hence, the protocol developed here enables highly sensitive thiol analysis in samples with complexity comparable to that expected in astrobiologically relevant extraterrestrial settings. This new protocol could be readily added to the existing suite of microfluidic chemical analyses developed for in situ planetary exploration; all that is required is the incorporation of two new reagents to the payload of an existing instrument concept. PMID:23161601

Mora, Maria F; Stockton, Amanda M; Willis, Peter A

2012-12-18

285

Thimerosal Exposure and the Role of Sulfation Chemistry and Thiol Availability in Autism  

Science.gov (United States)

Autism spectrum disorder (ASD) is a neurological disorder in which a significant number of the children experience a developmental regression characterized by a loss of previously acquired skills and abilities. Typically reported are losses of verbal, nonverbal, and social abilities. Several recent studies suggest that children diagnosed with an ASD have abnormal sulfation chemistry, limited thiol availability, and decreased glutathione (GSH) reserve capacity, resulting in a compromised oxidation/reduction (redox) and detoxification capacity. Research indicates that the availability of thiols, particularly GSH, can influence the effects of thimerosal (TM) and other mercury (Hg) compounds. TM is an organomercurial compound (49.55% Hg by weight) that has been, and continues to be, used as a preservative in many childhood vaccines, particularly in developing countries. Thiol-modulating mechanisms affecting the cytotoxicity of TM have been identified. Importantly, the emergence of ASD symptoms post-6 months of age temporally follows the administration of many childhood vaccines. The purpose of the present critical review is provide mechanistic insight regarding how limited thiol availability, abnormal sulfation chemistry, and decreased GSH reserve capacity in children with an ASD could make them more susceptible to the toxic effects of TM routinely administered as part of mandated childhood immunization schedules.

Kern, Janet K.; Haley, Boyd E.; Geier, David A.; Sykes, Lisa K.; King, Paul G.; Geier, Mark R.

2013-01-01

286

Biochemical Characterization of a Thiol-Activated, Oxidation Stable Keratinase from Bacillus pumilus KS12  

Digital Repository Infrastructure Vision for European Research (DRIVER)

An extracellular keratinase from Bacillus pumilus KS12 was purified by DEAE ion exchange chromatography. It was a 45?kDa monomer as determined by SDS PAGE analysis. It was found to be an alkaline, serine protease with pH and temperature optima of 10 and 60C?, respectively. It was thiol...

Rinky Rajput; Richa Sharma; Rani Gupta

287

Biochemical Characterization of a Thiol-Activated, Oxidation Stable Keratinase from Bacillus pumilus KS12  

Digital Repository Infrastructure Vision for European Research (DRIVER)

An extracellular keratinase from Bacillus pumilus KS12 was purified by DEAE ion exchange chromatography. It was a 45?kDa monomer as determined by SDS PAGE analysis. It was found to be an alkaline, serine protease with pH and temperature optima of 10 and 60°C, respectively. It was thiol activated wit...

Rajput, Rinky; Sharma, Richa; Gupta, Rani

288

Mercury and non-protein thiol compounds in the seagrass Posidonia oceanica.  

UK PubMed Central (United Kingdom)

Mercury concentrations, non-protein thiol levels and the enzyme activities of glutathione-S-transferase (GST) were measured in the blades and sheaths of the marine phanerogam Posidonia oceanica. The seagrass was collected in January and June and at three sites: the Bay of Rosignano (Italy) known for its mercury contamination, the north of the Lérins islands (Bay of Cannes, France), the Bay of Tonnara (Corsica, France). The two latter sites are considered as free of any known industrial inputs. Mercury concentrations and GST activities in both tissues were always higher in samples from Rosignano, particularly in June. Non-protein thiol levels were significantly higher in the blades than in the sheaths of P. oceanica from Tonnara and Lérins. In contrast, at Rosignano, the sheaths presented a significantly higher non-protein thiol concentration than the blades, particularly in June. Levels in the sheaths appeared to increase with the degree of pollution. Western Blot performed on sheaths of P. oceanica collected in June at Rosignano and Lérins revealed a characteristic band of GSTs at 31 kDa, proving the presence of the GST enzyme in this tissue. Mercury seemed to exert an influence upon non-protein thiol metabolism, including GST induction, in P. oceanica collected from the NW Mediterranean.

Ferrat L; Gnassia-Barelli M; Pergent-Martini C; Roméo M

2003-01-01

289

Functional monolayers on oxide-free silicon surfaces via thiol-ene click chemistry  

Digital Repository Infrastructure Vision for European Research (DRIVER)

Thiol–ene click chemistry was used for the attachment of a variety of functional molecules onto oxide-free Si(111) surfaces using very mild conditions; the efficient nature of this coupling strategy allowed for successful light-induced micropatterning and thus provides a novel route towards biofunct...

Caipa Campos, M.A.; Paulusse, J.M.J.; Zuilhof, H.

290

Thiol-Ene Induced Diphosphonic Acid Functionalization of Superparamagnetic Iron Oxide Nanoparticles  

Energy Technology Data Exchange (ETDEWEB)

Multi-functional organic molecules represent an interesting challenge for nanoparticle functionalization due to the potential for undesirable interactions between the substrate material and the variable functionalities, making it difficult to control the final orientation of the ligand. In the present study, UV-induced thiol-ene click chemistry has been utilized as a means of directed functionalization of bifunctional ligands on an iron oxide nanoparticle surface. Allyl diphosphonic acid ligand was covalently deposited on the surface of thiol-presenting iron oxide nanoparticles via the formation of a UV-induced thioether. This method of thiol-ene click chemistry offers a set of reaction conditions capable of controlling the ligand deposition and circumventing the natural affinity exhibited by the phosphonic acid moiety for the iron oxide surface. These claims are supported via a multimodal characterization platform which includes thermogravimetric analysis, x-ray photoelectron spectroscopy, and metal contact analysis and are consistent with a properly oriented, highly active ligand on the nanoparticle surface. These experiments suggest thiol-ene click chemistry as both a practical and generally applicable strategy for the directed deposition of multi-functional ligands on metal oxide nanoparticle surfaces.

Rutledge, Ryan D.; Warner, Cynthia L.; Pittman, Jonathan W.; Addleman, Raymond S.; Engelhard, Mark H.; Chouyyok, Wilaiwan; Warner, Marvin G.

2010-07-20

291

Identifying volatile thiols in natural and city gases using a portable, potentiometric instrument  

Energy Technology Data Exchange (ETDEWEB)

In order to support real time identification of the concentration of odorizing agents such as thiols added to fuel gases, potentiometric (mercury measuring) methods for inverse titration of gas samples are developed and a portable instrument which operates on this principle. The precision of determination was compared with adsorptive and gas chromatographic methods and is sufficient for industrial use.

Garai, T.; Daroczy, J.; Nadudvari, J.; Szucs, M.; Vasanits, D.

1983-01-01

292

Assemblies of thiol-capped nanocrystals as functional units for use in nanotechnology  

Digital Repository Infrastructure Vision for European Research (DRIVER)

This work summarizes results of about 10 years of the author’s own activities in the field of aqueous synthesis and handling of thiol-capped semiconductor nanocrystals. As this field has also been explored by hundreds of other scientists, I have endeavoured to do my utmost to provide a short but com...

Gaponik, Nikolai

293

Factors influencing the oxidation of cysteamine and other thiols: implications for hyperthermic sensitization and radiation protection  

Energy Technology Data Exchange (ETDEWEB)

Some of the factors influencing the oxygen uptake and peroxide formation for cysteamine (MEA) and other thiols in serum-supplemented modified McCoy's 5A, a well-known medium used to cultivate a variety of cells in vitro, have been studied. The oxidation of MEA and cysteine in modified McCoy's 5A has been compared with that in Ham's F-12, MEM, and phosphate-buffered saline. The ability to produce peroxide is dependent upon the temperature, the concentration of thiol, the presence of copper ions, and pH of the medium. Catalase also reduces the oxygen uptake for all thiols. Superoxide dismutase (SOD) was found to stimulate the oxygen uptake in the case of MEA and cysteine, but had little or no effect with DTT and glutathione. The combined presence of SOD and catalase resulted in less inhibition of oxygen uptake than that obtained by catalase alone. Alkaline pH was found to enhance the oxidation of cysteine and MEA. The results indicate that many problems may arise when thiols are added to various media. A major consideration is concerned with the production of peroxide, superoxide, and reduced trace metal intermediates. The presence of these intermediates may result in the production of hydroxyl radical intermediates as well as the eventual oxygen depletion from the medium.

Biaglow, J.E.; Issels, R.W.; Gerweck, L.E.; Varnes, M.E.; Jacobson, B.; Mittchell, J.B.; Russo, A.

1984-11-01

294

Protective effects of thiol compounds on chromate-induced toxicity in vitro and in vivo.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

The effects of thiol compounds (L-cysteine ethyl ester, 2,3-dimercaptosuccinic acid, or 2,3-dimercapto-1-propanesulfonic acid) on the toxicity induced by chromate (potassium dichromate) were investigated in HeLa cells and mice. Chromate-induced cytotoxicity evaluated by inhibition of cell growth and...

Susa, N; Ueno, S; Furukawa, Y

295

Aroma extraction dilution analysis of Sauternes wines. Key role of polyfunctional thiols.  

Science.gov (United States)

The aim of the present work was to investigate Sauternes wine aromas. In all wine extracts, polyfunctional thiols were revealed to have a huge impact. A very strong bacon-petroleum odor emerged at RI = 845 from a CP-Sil5-CB column. Two thiols proved to participate in this perception: 3-methyl-3-sulfanylbutanal and 2-methylfuran-3-thiol. A strong synergetic effect was evidenced between the two compounds. The former, never mentioned before in wines, and not found in the musts of this study, is most probably synthesized during fermentation. 3-Methylbut-2-ene-1-thiol, 3-sulfanylpropyl acetate, 3-sulfanylhexan-1-ol, and 3-sulfanylheptanal also contribute to the global aromas of Sauternes wines. Among other key odorants, the presence of a varietal aroma (alpha-terpineol), sotolon, fermentation alcohols (3-methylbutan-1-ol and 2-phenylethanol) and esters (ethyl butyrate, ethyl hexanoate, and ethyl isovalerate), carbonyls (trans-non-2-enal and beta-damascenone), and wood flavors (guaiacol, vanillin, eugenol, beta-methyl-gamma-octalactone, and Furaneol) is worth stressing. PMID:16968087

Bailly, Sabine; Jerkovic, Vesna; Marchand-Brynaert, Jacqueline; Collin, Sonia

2006-09-20

296

Thiol-ene click chemistry for the synthesis of highly effective glycosyl sulfonamide carbonic anhydrase inhibitors.  

UK PubMed Central (United Kingdom)

Thiol-ene click chemistry has been applied for obtaining sulfonamide carbonic anhydrase (CA, EC 4.2.1.1) inhibitors incorporating sugar moieties. Most of these new compounds were moderate CA I inhibitors, effective CA II inhibitors, and low nanomolar/subnanomolar inhibitors of the tumor-associated isoforms CA IX and XII.

Saada MC; Ombouma J; Montero JL; Supuran CT; Winum JY

2013-06-01

297

Synthesis and Microstructural Investigations of Organometallic Pd(II) Thiol-Gold Nanoparticles Hybrids  

Directory of Open Access Journals (Sweden)

Full Text Available Abstract In this work the synthesis and characterization of gold nanoparticles functionalized by a novel thiol-organometallic complex containing Pd(II) centers is presented. Pd(II) thiol,trans, trans-[dithiolate-dibis(tributylphosphine)dipalladium(II)-4,4?-diethynylbiphenyl] was synthesized and linked to Au nanoparticles by the chemical reduction of a metal salt precursor. The new hybrid made of organometallic Pd(II) thiol-gold nanoparticles, shows through a single S bridge a direct link between Pd(II) and Au nanoparticles. The size-control of the Au nanoparticles (diameter range 2–10 nm) was achieved by choosing the suitable AuCl4 ?/thiol molar ratio. The size, strain, shape, and crystalline structure of these functionalized nanoparticles were determined by a full-pattern X-ray powder diffraction analysis, high-resolution TEM, and X-ray photoelectron spectroscopy. Photoluminescence spectroscopy measurements of the hybrid system show emission peaks at 418 and 440 nm. The hybrid was exposed to gaseous NO x with the aim to evaluate the suitability for applications in sensor devices; XPS measurements permitted to ascertain and investigate the hybrid –gas interaction.

Vitale Floriana; Vitaliano Rosa; Battocchio Chiara; Fratoddi Ilaria; Giannini Cinzia; Piscopiello Emanuela; Guagliardi Antonella; Cervellino Antonio; Polzonetti Giovanni; Russo MariaVittoria; Tapfer Leander

2008-01-01

298

A polymer network prepared by the thiol-yne photocrosslinking of a liquid crystalline dendrimer.  

UK PubMed Central (United Kingdom)

A polymer network is prepared by the thiol-yne photopolymerization of multifunctional dendrimer with a tetrathiol crosslinker. The network obtained shows a liquid crystalline phase at room temperature, which has been characterized by optical microscopy, differential scanning calorimetry, and X-ray diffraction. Photoinduced deformation of uniaxially aligned free-standing films of the photocrosslinked material has been demonstrated.

Cervera-Procas R; Sánchez-Somolinos C; Serrano JL; Omenat A

2013-03-01

299

A polymer network prepared by the thiol-yne photocrosslinking of a liquid crystalline dendrimer.  

Science.gov (United States)

A polymer network is prepared by the thiol-yne photopolymerization of multifunctional dendrimer with a tetrathiol crosslinker. The network obtained shows a liquid crystalline phase at room temperature, which has been characterized by optical microscopy, differential scanning calorimetry, and X-ray diffraction. Photoinduced deformation of uniaxially aligned free-standing films of the photocrosslinked material has been demonstrated. PMID:23322378

Cervera-Procas, Ramón; Sánchez-Somolinos, Carlos; Serrano, José L; Omenat, Ana

2013-01-16

300

Change in plasma and erythrocyte thiol levels in children undergoing fasting studies for investigation of hypoglycaemia  

Directory of Open Access Journals (Sweden)

Full Text Available Introduction: It is unclear how repeated episodes of HG cause brain injury, but oxidative stress has been suggested to play a role. Non-protein thiols (glutathione, GSH, and cysteine, CYSH) form an important antioxidant defence, and their redox state is dependent on intracellular energy supplies and reducing power which are possibly compromised during low blood glucose concentrations. Aim of study: To study thiol status in children undergoing investigations for hypoglycaemia . We hypothesised that thiol metabolism, dependent on intracellular energy supplies and reducing power, might deteriorate during hypoglycaemic episodes. Material and methods: Seventeen children with suspected hypoglycaemic episodes underwent a diagnostic fast. We measured plasma and erythrocyte GSH and CYSH as well as activities of enzymes related to glutathione metabolism during and after the fast. Results: A positive correlation between plasma glucose and reduced (free) cysteine was observed, but plasma GSH levels did not change significantly. Hypoglycaemia was associated with a rise in erythrocyte total cysteine without changes in erythrocyte GSH. Conclusion: Low blood glucose concentration is associated with changes in thiol status, which compromise the protection against oxidative stress. This further supports the hypothesis of oxidative stress being associated with hypoglycaemia.

Heli Salmi; Khalid Hussain; Risto Lapatto

2011-01-01

 
 
 
 
301

An experimental comparison of Thiol broth with Brewer's thioglycollate for anaerobic blood cultures.  

Digital Repository Infrastructure Vision for European Research (DRIVER)

In a series of simulated blood culture experiments, small inocula of eight different strains of Bacteroides and five strains of anaerobic cocci were added to Difco Thiol broth and Southern Group Brewer's thioglycollate. Both methods enabled all of the strains to be isolated after one to three days' ...

Shanson, D C; Barnicoat

302

Ribonucleotide reductase association with mammalian liver mitochondria.  

Science.gov (United States)

Deoxyribonucleoside triphosphate pools in mammalian mitochondria are highly asymmetric, and this asymmetry probably contributes to the elevated mutation rate for the mitochondrial genome as compared with the nuclear genome. To understand this asymmetry, we must identify pathways for synthesis and accumulation of dNTPs within mitochondria. We have identified ribonucleotide reductase activity specifically associated with mammalian tissue mitochondria. Examination of immunoprecipitated proteins by mass spectrometry revealed R1, the large ribonucleotide reductase subunit, in purified mitochondria. Significant enzymatic and immunological activity was seen in rat liver mitochondrial nucleoids, isolated as described by Wang and Bogenhagen (Wang, Y., and Bogenhagen, D. F. (2006) J. Biol. Chem. 281, 25791-25802). Moreover, incubation of respiring rat liver mitochondria with [(14)C]cytidine diphosphate leads to accumulation of radiolabeled deoxycytidine and thymidine nucleotides within the mitochondria. Comparable results were seen with [(14)C]guanosine diphosphate. Ribonucleotide reduction within the mitochondrion, as well as outside the organelle, needs to be considered as a possibly significant contributor to mitochondrial dNTP pools. PMID:23504325

Chimploy, Korakod; Song, Shiwei; Wheeler, Linda J; Mathews, Christopher K

2013-03-15

303

Synthesis of Hyperbranched Polypeptide and PEO Block Copolymer by Consecutive Thiol-Yne Chemistry.  

UK PubMed Central (United Kingdom)

Hyperbranched poly(?-benzyloxycarbonyl-l-lysine) (HPlys) with multiple alkyne peripheries was synthesized through the click polycondensation of an AB2 type Plys macromonomer with ?-thiol and ?-alkyne terminal groups (thiol is the A unit, and each ? bond in alkyne is the B unit), and the resulting HPlys was further conjugated with thiol-termined poly(ethylene oxide) (PEO) to generate HPlys-b-PEO block copolymer by consecutive thiol-yne chemistry. Their molecular structures and physical properties were characterized in detail by FT-IR, (1)H NMR, gel permeation chromatography, differential scanning calorimetry, wide-angle X-ray diffraction, and polarized optical microscopy. HPlys and HPlys-b-PEO mainly assumed an ?-helix conformation similar to the linear precursors, while the liquid crystalline phase transition of Plys segment disappeared within HPlys and HPlys-b-PEO. HPlys-b-PEO self-assembled into nearly spherical micelles in aqueous solution, while it gave a 5-fold lower critical aggregation concentration (8.9 × 10(-3) mg/mL) than a linear counterpart (4.5 × 10(-2) mg/mL), demonstrating a dendritic topology effect. Compared with a linear counterpart, HPlys-b-PEO gave a higher drug-loading capacity and efficiency for the anticancer drug doxorubicin (DOX) and a slower drug-release rate with an improved burst-release profile, enabling them useful for drug delivery systems. Importantly, this work provides a versatile strategy for the synthesis of hyperbranched polypeptides and related block copolymers by utilizing thiol-yne chemistry.

Chang X; Dong CM

2013-09-01

304

Synthesis of Hyperbranched Polypeptide and PEO Block Copolymer by Consecutive Thiol-Yne Chemistry.  

Science.gov (United States)

Hyperbranched poly(?-benzyloxycarbonyl-l-lysine) (HPlys) with multiple alkyne peripheries was synthesized through the click polycondensation of an AB2 type Plys macromonomer with ?-thiol and ?-alkyne terminal groups (thiol is the A unit, and each ? bond in alkyne is the B unit), and the resulting HPlys was further conjugated with thiol-termined poly(ethylene oxide) (PEO) to generate HPlys-b-PEO block copolymer by consecutive thiol-yne chemistry. Their molecular structures and physical properties were characterized in detail by FT-IR, (1)H NMR, gel permeation chromatography, differential scanning calorimetry, wide-angle X-ray diffraction, and polarized optical microscopy. HPlys and HPlys-b-PEO mainly assumed an ?-helix conformation similar to the linear precursors, while the liquid crystalline phase transition of Plys segment disappeared within HPlys and HPlys-b-PEO. HPlys-b-PEO self-assembled into nearly spherical micelles in aqueous solution, while it gave a 5-fold lower critical aggregation concentration (8.9 × 10(-3) mg/mL) than a linear counterpart (4.5 × 10(-2) mg/mL), demonstrating a dendritic topology effect. Compared with a linear counterpart, HPlys-b-PEO gave a higher drug-loading capacity and efficiency for the anticancer drug doxorubicin (DOX) and a slower drug-release rate with an improved burst-release profile, enabling them useful for drug delivery systems. Importantly, this work provides a versatile strategy for the synthesis of hyperbranched polypeptides and related block copolymers by utilizing thiol-yne chemistry. PMID:23957555

Chang, Xiao; Dong, Chang-Ming

2013-08-27

305

Low amounts and high thiol oxidation of peroxiredoxins in spermatozoa from infertile men.  

UK PubMed Central (United Kingdom)

Seminal oxidative stress occurs when there is an increased production of reactive oxygen species (ROS) and/or a decrease of antioxidant activity, promoting impaired sperm function. Peroxiredoxins (PRDX) are abundant in human semen and are important antioxidant enzymes, which act as ROS scavengers and modulators in ROS-dependent signaling. Our aim was to determine whether the levels of PRDX1 and PRDX6 and their oxidation on thiol groups are associated with a decrease in sperm motility and DNA integrity. We evaluated the sperm and seminal PRDX level in men (13 healthy controls, 15 men with clinical varicocele, and 17 men with idiopathic infertility). We assessed conventional semen parameters, sperm DNA integrity (by the sperm chromatin structure assay), lipid peroxidation in seminal plasma and spermatozoa (by the thiobarbituric acid reactive substances assay), and the amount and thiol oxidation of PRDX1 and PRDX6 (by immunoblotting). PRDXs were affected in seminal plasma (lower amounts) and in sperm samples (lower amounts and higher levels of thiol oxidation) characterized by lower sperm motility, higher lipid peroxidation, and sperm DNA damage. The thioloxidation ratio of PRDXs (thiol-oxidized PRDX/total PRDX) correlated negatively with sperm motility (total and progressive) and positively with sperm DNA damage and sperm lipid peroxidation. In conclusion, because of the lower amount of total PRDX1 and PRDX6 and the high thiol oxidation of these PRDXs, very little (less than 20%) protection due to PRDXs remains, and this is associated with impaired sperm function and poor DNA integrity and suggests an important role of PRDXs in the protection of human spermatozoa against oxidative stress.

Gong S; San Gabriel MC; Zini A; Chan P; O'Flaherty C

2012-11-01

306

Chromenoquinoline-based thiol probes: a study on the quencher position for controlling fluorescent Off-On characteristics.  

UK PubMed Central (United Kingdom)

The design, synthesis and thiol sensing ability of chromenoquinoline-based fluorescent probes 4, 5 and 6 and are reported here. The relative position of the maleimide moiety was varied along the chromenoquinoline fluorophore to decrease the background fluorescence. Lower background fluorescence in probes 4 and 6 was rationalized by the smaller k(r)/k(nr) values compared to that of probe 5. An intramolecular charge transfer (ICT) mechanism was proposed for quenching and the extent was dependent on the position of the maleimide quencher. Fluorescent Off-On characteristics were evaluated by theoretical calculations. All probes were selective only towards thiol containing amino acids. Thiol sensing by probes 4 and 6 were much better compared to 5. Probe 4 displayed a better fluorescence response for less hindered thiol (185-, 223- and 156-fold for Hcy, Cys and GSH, respectively), while for probe 6, a higher enhancement in fluorescence was observed with more hindered thiols (180-, 205- and 245-fold for Hcy, Cys and GSH, respectively). The better response to bulkier thiol, GSH by probe 6 was attributed to the steric crowding at the C-4 position and bulkiness of the GSH group which force the succinimide unit to be in a nearly orthogonal conformation. This spatial arrangement was important in reducing the fluorescence quenching ability of the succinimide moiety. The application of probes 4, 5 and 6 was demonstrated by naked eye detection thiols using a 96-well plate system as well as by live-cell imaging.

Kand D; Kalle AM; Talukdar P

2013-02-01

307

Comparison of thiol subproteome of the vent mussel Bathymodiolus azoricus from different Mid-Atlantic Ridge vent sites.  

UK PubMed Central (United Kingdom)

Deep-sea hydrothermal mussels Bathymodiolus azoricus live in the mixing zone where hydrothermal fluid mixes with bottom seawater, creating large gradients in the environmental conditions and are one of the most studied hydrothermal species as a model of adaptation to extreme conditions. Thiol proteins, i.e. proteins containing a thiol or sulfhydryl group (SH) play major roles in intracellular stress defense against reactive oxygen species (ROS) and are especially susceptible to oxidation. However, they are not particularly abundant, representing a small percentage of proteins in the total proteome and therefore are difficult to study by proteomic approaches. Activated thiol sepharose (ATS) was used for the rapid and quantitative selection of proteins comprising thiol- or disulfide-containing subproteomes. This study aims to isolate thiol-containing proteins from the gills of B. azoricus collected in distinct hydrothermal vents and to study the thiol-containing subproteome as a function of site-specific susceptibility to ROS. Results show that ATS is a powerful tool to isolate the thiol-containing sub-proteome and differently-expressed protein spots showed significant differences among the three vent sites, supporting previous findings that specific environmental conditions are crucial for ROS formation and that B. azoricus have different susceptibilities to oxidative stress depending on the vent site they inhabit.

Company R; Torreblanca A; Cajaraville M; Bebianno MJ; Sheehan D

2012-10-01

308

Formation of Underbrushes on thiolated Poly (ethylene glycol) PEG monolayers by Oligoethylene glycol (OEG) terminated Alkane Thiols on Gold  

DEFF Research Database (Denmark)

Adding underbrushes of oligoethylene glycol (OEG) to monolayers of long chain PEG molecules on a surface is one of the strategies [1] in designing a suitable platform for antifouling purpose, where it is possible to have high graft density and molecular conformational freedom[4] simultaneously, there by maximal retention of activity of covalently immobilised antifouling enzyme [2] on PEG surfaces along with resistance to protein adsorption[3]. Here we present some our studies on the addition of OEG thiol molecules over a self assembled monolayer of PEG thiol on gold. The kinetics of addition of OEG thiol to monolayers of PEG thiol was followed using X- ray photoelectron spectroscopy (XPS), which indicated the time point of maximum graft density and beyond this time point there was predominant desorption of OEG thiol as indicated by the C/O ratio. The initial increase in graft density was reflected in the superior resistance towards non specific adsorption of proteins as shown by N 1s signal. We also performedprotein adsorption studies using quartz crystal microbalance (QCM-D). Studies involving addition of alkane thiol instead of OEG terminating alkane thiol showed the importance of OEG part of the molecule in superior resistance towards protein adsorption. The surfaces with underbrushes were imaged using atomic force microscopy (AFM) to detect any changes in mechanical properties of PEG thiol covered surfaces upon addition of OEG thiol. References: 1. Katsumi Uchida, Yuki Hoshino, Atsushi Tamura, Keitaro Yoshimoto, Shuji Kojima and Keichiro Yamashita, Ichiro Yamanaka, Hidenori Otsuka, Kazunori Kataoka, Yukio Nagasaki, Biointerphases. 2007, 2, 4, 126. 2. L. Selan, F. Berluti, C. Passariello, M. R. Comodiballanti, M. C. Thaller, Antimicrobial agents and chemotherapy, 1993, 37, 12, 2618. 3. Susan J. Sofia, V. Premnath, and Edward W. Merrill, Macromolecules, 1998, 31, 15, 5059. 4. Hidenori Otsuka, Yukio Nagasaki, and Kazunori Kataoka, Langmuir, 2004, 20, 26, 11285

Lokanathan, Arcot R.

2011-01-01

309

A high-throughput assay format for determination of nitrate reductase and nitrite reductase enzyme activities  

Energy Technology Data Exchange (ETDEWEB)

The authors describe a microplate-based high-throughput procedure for rapid assay of the enzyme activities of nitrate reductase and nitrite reductase, using extremely small volumes of reagents. The new procedure offers the advantages of rapidity, small sample size-nanoliter volumes, low cost, and a dramatic increase in the throughput sample number that can be analyzed simultaneously. Additional advantages can be accessed by using microplate reader application software packages that permit assigning a group type to the wells, recording of the data on exportable data files and exercising the option of using the kinetic or endpoint reading modes. The assay can also be used independently for detecting nitrite residues/contamination in environmental/food samples. 10 refs., 2 figs.

McNally, N.; Liu, Xiang Yang; Choudary, P.V. [Univ. of California, Davis, CA (United States)

1997-01-01

310

Hydroxylated naphthoquinones as substrates for Escherichia coli anaerobic reductases.  

Science.gov (United States)

We have used two hydroxylated naphthoquinol menaquinol analogues, reduced plumbagin (PBH2, 5-hydroxy-2-methyl-1,4-naphthoquinol) and reduced lapachol [LPCH2, 2-hydroxy-3-(3-methyl-2-butenyl)-1, 4-naphthoquinol], as substrates for Escherichia coli anaerobic reductases. These compounds have optical, solubility and redox properties that make them suitable for use in studies of the enzymology of menaquinol oxidation. Oxidized plumbagin and oxidized lapachol have well resolved absorbances at 419 nm (epsilon=3.95 mM-1. cm-1) and 481 nm (epsilon=2.66 mM-1.cm-1) respectively (in Mops/KOH buffer, pH 7.0). PBH2 is a good substrate for nitrate reductase A (Km=282+/-28 microM, kcat=120+/-6 s-1) and fumarate reductase (Km=155+/-24 microM, kcat=30+/-2 s-1), but not for DMSO reductase. LPCH2 is a good substrate for nitrate reductase A (Km=57+/-35 microM, kcat=68+/-13 s-1), fumarate reductase (Km=85+/-27 microM, kcat=74+/-6 s-1) and DMSO reductase (Km=238+/-30 microM, kcat=191+/-21 s-1). The sensitivity of enzymic LPCH2 and PBH2 oxidation to 2-n-heptyl-4-hydroxyquinoline N-oxide inhibition is consistent with their oxidation occurring at sites of physiological quinol binding.

Rothery, R A; Chatterjee, I; Kiema, G; McDermott, M T; Weiner, J H

1998-01-01

311

Porcine carbonyl reductase. structural basis for a functional monomer in short chain dehydrogenases/reductases.  

UK PubMed Central (United Kingdom)

Porcine testicular carbonyl reductase (PTCR) belongs to the short chain dehydrogenases/reductases (SDR) superfamily and catalyzes the NADPH-dependent reduction of ketones on steroids and prostaglandins. The enzyme shares nearly 85% sequence identity with the NADPH-dependent human 15-hydroxyprostaglandin dehydrogenase/carbonyl reductase. The tertiary structure of the enzyme at 2.3 A reveals a fold characteristic of the SDR superfamily that uses a Tyr-Lys-Ser triad as catalytic residues, but exhibits neither the functional homotetramer nor the homodimer that distinguish all SDRs. It is the first known monomeric structure in the SDR superfamily. In PTCR, which is also active as a monomer, a 41-residue insertion immediately before the catalytic Tyr describes an all-helix subdomain that packs against interfacial helices, eliminating the four-helix bundle interface conserved in the superfamily. An additional anti-parallel strand in the PTCR structure also blocks the other strand-mediated interface. These novel structural features provide the basis for the scaffolding of one catalytic site within a single molecule of the enzyme.

Ghosh D; Sawicki M; Pletnev V; Erman M; Ohno S; Nakajin S; Duax WL

2001-05-01

312

Mechanism of thiol-supported arsenate reduction mediated by phosphorolytic-arsenolytic enzymes: II. Enzymatic formation of arsenylated products susceptible for reduction to arsenite by thiols.  

Science.gov (United States)

Enzymes catalyzing the phosphorolytic cleavage of their substrates can reduce arsenate (AsV) to the more toxic arsenite (AsIII) via the arsenolytic substrate cleavage in presence of a reductant, as glutathione or dithiotreitol (DTT). We have shown this for purine nucleoside phosphorylase (PNP), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), glycogen phosphorylase-a (GPa), and phosphotransacetylase (PTA). Using a multidisciplinary approach, we explored the mechanism whereby these enzymes mediate AsV reduction. It is known that PNP cleaves inosine with AsV into hypoxanthine and ribose-1-arsenate. In presence of inosine, AsV and DTT, PNP mediates AsIII formation. In this study, we incubated PNP first with inosine and AsV, allowing the arsenolytic reaction to run, then blocked this reaction with the PNP inhibitor BCX-1777, added DTT and continued the incubation. Despite inhibition of PNP, large amount of AsIII was formed in these incubations, indicating that PNP does not reduce AsV directly but forms a product (i.e., ribose-1-arsenate) that is reduced to AsIII by DTT. Similar studies with the other arsenolytic enzymes (GPa, GAPDH, and PTA) yielded similar results. Various thiols that differentially supported AsV reduction when present during PNP-catalyzed arsenolysis (DTT approximately dimercaptopropane-1-sulfonic acid > mercaptoethanol > DMSA > GSH) similarly supported AsV reduction when added only after a transient PNP-catalyzed arsenolysis, which preformed ribose-1-arsenate. Experiments with progressively delayed addition of DTT after BCX-1777 indicated that ribose-1-arsenate is short-lived with a half-life of 4 min. In conclusion, phosphorolytic enzymes, such as PNP, GAPDH, GPa, and PTA, promote thiol-dependent AsV reduction because they convert AsV into arsenylated products reducible by thiols more readily than AsV. In support of this view, reactivity studies using conceptual density functional theory reactivity descriptors (local softness, nucleofugality) indicate that reduction by thiols of the arsenylated metabolites is favored over AsV. PMID:19478237

Gregus, Zoltán; Roos, Goedele; Geerlings, Paul; Németi, Balázs

2009-05-28

313

Methylenetetrahydrofolate reductase variant and schizophrenia/depression.  

UK PubMed Central (United Kingdom)

Patients with methylenetetrahydrofolate reductase (MTHFR) deficiency often show psychiatric manifestations. Since a common variant of the MTHFR gene, T677(Ala), responsible for the thermolabile MTHFR with less than 50% specific MTHFR activity, has been reported, we examined whether the T677 allele is associated with psychiatric disorders in an unrelated Japanese population consisting of 297 schizophrenics, 32 patients with major depression, 40 patients with bipolar disorder, and 419 controls. The genotype homozygous for the T677 allele was significantly frequently observed in schizophrenics with an odds ratio of 1.9 (P = 0.0006), and in patients with major depression with an odds ratio of 2.8 (P = 0.005). Our data suggest associations of the MTHFR gene variant with schizophrenia and depression in the Japanese.

Arinami T; Yamada N; Yamakawa-Kobayashi K; Hamaguchi H; Toru M

1997-09-01

314

Methylenetetrahydrofolate reductase variant and schizophrenia/depression.  

Science.gov (United States)

Patients with methylenetetrahydrofolate reductase (MTHFR) deficiency often show psychiatric manifestations. Since a common variant of the MTHFR gene, T677(Ala), responsible for the thermolabile MTHFR with less than 50% specific MTHFR activity, has been reported, we examined whether the T677 allele is associated with psychiatric disorders in an unrelated Japanese population consisting of 297 schizophrenics, 32 patients with major depression, 40 patients with bipolar disorder, and 419 controls. The genotype homozygous for the T677 allele was significantly frequently observed in schizophrenics with an odds ratio of 1.9 (P = 0.0006), and in patients with major depression with an odds ratio of 2.8 (P = 0.005). Our data suggest associations of the MTHFR gene variant with schizophrenia and depression in the Japanese. PMID:9342205

Arinami, T; Yamada, N; Yamakawa-Kobayashi, K; Hamaguchi, H; Toru, M

1997-09-19

315

Methylenetetrahydrofolate reductase: biochemical characterization and medical significance.  

UK PubMed Central (United Kingdom)

Methylenetetrahydrofolate reductase (MTHFR) catalyzes the reduction of 5,10-methylenetetrahydofolate (CH2-H4folate) to 5-methyltetrahydrofolate (CH3-H4folate). The enzyme employs a noncovalently-bound flavin adenine dinucleotide (FAD), which accepts reducing equivalents from NAD(P)H and transfers them to CH2-H4folate. The reaction provides the sole source of CH3-H4folate, which is utilized by methionine synthase in the synthesis of methionine from homocysteine. MTHFR plays a key role in folate metabolism and in the homeostasis of homocysteine; mutations in the enzyme lead to hyperhomocyst(e)inemia. A common C677T polymorphism in MTHFR has been associated with an increased risk for the development of cardiovascular disease, Alzheimer's disease, and depression in adults, and of neural tube defects in the fetus. The mutation also confers protection for certain types of cancers. This review presents the current knowledge of the enzyme, its biochemical characterization, and medical significance.

Trimmer EE

2013-01-01

316

Dynamics of trimethoprim bound to dihydrofolate reductase  

International Nuclear Information System (INIS)

The conformation of a small molecule in its binding site on a protein is a major factor in the specificity of the interaction between them. In this paper, the authors report the use of 1H and 13C NMR spectroscopy to study the fluctuations in conformation of the anti-bacterial drug trimethoprim when it is bound to its target, dihydrofolate reductase. 13C relaxation measurements reveal dihedral angle changes of ±25 degree to ±35 degree on the subnanosecond time scale, while 13C line-shape analysis demonstrates dihedral angle changes of at least ±65 degree on the millisecond time scale. 1H NMR shows that a specific hydrogen bond between the inhibitor and enzyme, which is believed to make an important contribution to binding, makes and breaks rapidly at room temperature.

1988-01-01

317

The cytochrome bd respiratory oxygen reductases.  

UK PubMed Central (United Kingdom)

Cytochrome bd is a respiratory quinol: O? oxidoreductase found in many prokaryotes, including a number of pathogens. The main bioenergetic function of the enzyme is the production of a proton motive force by the vectorial charge transfer of protons. The sequences of cytochromes bd are not homologous to those of the other respiratory oxygen reductases, i.e., the heme-copper oxygen reductases or alternative oxidases (AOX). Generally, cytochromes bd are noteworthy for their high affinity for O? and resistance to inhibition by cyanide. In E. coli, for example, cytochrome bd (specifically, cytochrome bd-I) is expressed under O?-limited conditions. Among the members of the bd-family are the so-called cyanide-insensitive quinol oxidases (CIO) which often have a low content of the eponymous heme d but, instead, have heme b in place of heme d in at least a majority of the enzyme population. However, at this point, no sequence motif has been identified to distinguish cytochrome bd (with a stoichiometric complement of heme d) from an enzyme designated as CIO. Members of the bd-family can be subdivided into those which contain either a long or a short hydrophilic connection between transmembrane helices 6 and 7 in subunit I, designated as the Q-loop. However, it is not clear whether there is a functional consequence of this difference. This review summarizes current knowledge on the physiological functions, genetics, structural and catalytic properties of cytochromes bd. Included in this review are descriptions of the intermediates of the catalytic cycle, the proposed site for the reduction of O?, evidence for a proton channel connecting this active site to the bacterial cytoplasm, and the molecular mechanism by which a membrane potential is generated.

Borisov VB; Gennis RB; Hemp J; Verkhovsky MI

2011-11-01

318

Hexaheme nitrite reductase from Desulfovibrio desulfuricans  

Energy Technology Data Exchange (ETDEWEB)

Moessbauer and EPR spectroscopy were used to characterize the heme prosthetic groups of the nitrite reductase isolated from Desulfovibrio desulfuricans (ATCC 27774), which is a membrane-bound multiheme cytochrome capable of catalyzing the 6-electron reduction of nitrite to ammonia. At pH 7.6, the as-isolated enzyme exhibited a complex EPR spectrum consisting of a low-spin ferric heme signal at g = 2.96, 2.28, and 1.50 plus several broad resonances indicative of spin-spin interactions among the heme groups. EPR redox titration studies revealed yet another low-spin ferric heme signal at g = 3.2 and 2.14 (the third g value was undetected) and the presence of a high-spin ferric heme. Moessbauer measurements demonstrated further that this enzyme contained six distinct heme groups: one high-spin (S = 5/2) and five low-spin (S = 1/2) ferric hemes. Characteristic hyperfine parameters for all six hemes were obtained through a detailed analysis of the Moessbauer spectra. D. desulfuricans nitrite reductase can be reduced by chemical reductants, such as dithionite or reduced methyl viologen, or by hydrogenase under hydrogen atmosphere. Addition of nitrite to the fully reduced enzyme reoxidized all five low-spin hemes to their ferric states. The high-spin heme, however, was found to complex NO, suggesting that the high-spin heme could be the substrate binding site and that NO could be an intermediate present in an enzyme-bound form.

Costa, C.; Moura, J.J.G.; Moura, I. (Centro de Tecnologia Quimica e Biologica, Oeiras (Portugal) Univ. Nova de Lisboa, Oeiras (Portugal)); Liu, M.Y.; Peck, H.D. Jr.; LeGall, J. (Univ. of Georgia, Athens (United States)); Wang, Yaning; Huynh, B.H. (Emory Univ., Atlanta, GA (United States))

1990-08-25

319

Carboxylation mechanism and stereochemistry of crotonyl-CoA carboxylase/reductase, a carboxylating enoyl-thioester reductase  

Science.gov (United States)

Chemo- and stereoselective reductions are important reactions in chemistry and biology, and reductases from biological sources are increasingly applied in organic synthesis. In contrast, carboxylases are used only sporadically. We recently described crotonyl-CoA carboxylase/reductase, which catalyzes the reduction of (E)-crotonyl-CoA to butyryl-CoA but also the reductive carboxylation of (E)-crotonyl-CoA to ethylmalonyl-CoA. In this study, the complete stereochemical course of both reactions was investigated in detail. The pro-(4R) hydrogen of NADPH is transferred in both reactions to the re face of the C3 position of crotonyl-CoA. In the course of the carboxylation reaction, carbon dioxide is incorporated in anti fashion at the C2 atom of crotonyl-CoA. For the reduction reaction that yields butyryl-CoA, a solvent proton is added in anti fashion instead of the CO2. Amino acid sequence analysis showed that crotonyl-CoA carboxylase/reductase is a member of the medium-chain dehydrogenase/reductase superfamily and shares the same phylogenetic origin. The stereospecificity of the hydride transfer from NAD(P)H within this superfamily is highly conserved, although the substrates and reduction reactions catalyzed by its individual representatives differ quite considerably. Our findings led to a reassessment of the stereospecificity of enoyl(-thioester) reductases and related enzymes with respect to their amino acid sequence, revealing a general pattern of stereospecificity that allows the prediction of the stereochemistry of the hydride transfer for enoyl reductases of unknown specificity. Further considerations on the reaction mechanism indicated that crotonyl-CoA carboxylase/reductase may have evolved from enoyl-CoA reductases. This may be useful for protein engineering of enoyl reductases and their application in biocatalysis.

Erb, Tobias J.; Brecht, Volker; Fuchs, Georg; Muller, Michael; Alber, Birgit E.

2009-01-01

320

Voltammetry and Electrocatalysis of Achrornobacter Xylosoxidans Copper Nitrite Reductase on Functionalized Au(111)-Electrode Surfaces  

DEFF Research Database (Denmark)

A long-standing issue in protein film voltammetry (PFV), particularly electrocatalytic voltammetry of redox enzyme monolayers, is the variability of protein adsorption modes, reflected in distributions of catalytic activity of the adsorbed protein/enzyme molecules. Use of well-defined, atomically planar electrode surfaces is a step towards the resolution of this central issue. We report here the voltammetry of copper nitrite reductase (CNiR, Achromobacter xylosoxidons) on Au(111)-electrode surfaces modified by monolayers of a broad variety of thiol-based linker molecules. These represent positively charged and electrostatically neutral, hydrophobic and hydrophilic, aliphatic and aromatic, and variable-length micro-environments, as well as their combinations. Optimal conditions for enzyme function seems to be a combination of hydrophobic and hydrophilic surface linker properties, which can lead to close to complete non-catalytic monolayer interfacial electron transfer function and electrocatalysis with activity approaching enzyme activity in homogeneous solution. Thiophenol (combined hydrophobic stacking and interdispersed water molecules), 4-methyl-thiophenol (hydrophobic and water molecules), and 3- and 4-aminothiophenol(hydrophilic, hydrophobic) offer the overall most efficient micro-environments. Subtle differences with even small structural linker differences, however, lead to widely different electrocatalytic properties, strikingly illuminated by the (omega-mercaptoamines. CuNiR thus shows highly efficient, close to ideal reversible electrocatalytic voltammetry on cysteamine-covered Au(111)-electrode surfaces, most likely due to two cysteamine orientations previously disclosed by in situ scanning tunnelling microscopy. Such a dual orientation exposes both a hydrophobic and a positively charged, hydrophilic surface feature. In contrast, the higher cysteamine homologues expose only the hydrophilic component with no electrocatalytic activity on these surfaces. These results offer a basis for rational surface design in forthcoming biological electrocatalysis useful both fundamentally and in novel biosensor technology.

Welinder, Anna C.; Zhang, Jingdong

2007-01-01

 
 
 
 
321

Glutathione and glutathione reductase: a boon in disguise for plant abiotic stress defense operations.  

UK PubMed Central (United Kingdom)

Abiotic stresses such as salinity, drought, clilling, heavy metal are the major limiting factors for crop productivity. These stresses induce the overproduction of reactive oxygen species (ROS) which are highly reactive and toxic, which must be minimized to protect the cell from oxidative damage. The cell organelles, particularly chloroplast and mitochondria are the major sites of ROS production in plants where excessive rate of electron flow takes place. Plant cells are well equipped to efficiently scavenge ROS and its reaction products by the coordinated and concerted action of antioxidant machinery constituted by vital enzymatic and non-enzymatic antioxidant components. Glutathione reductase (GR, EC 1.6.4.2) and tripeptide glutathione (GSH, ?-Glutamyl-Cysteinyl-Glycine) are two major components of ascorbate-glutathione (AsA-GSH) pathway which play significant role in protecting cells against ROS and its reaction products-accrued potential anomalies. Both GR and GSH are physiologically linked together where, GR is a NAD(P)H-dependent enzymatic antioxidant and efficiently maintains the reduced pool of GSH - a cellular thiol. The differential modulation of both GR and GSH in plants has been widely implicated for the significance of these two enigmatic antioxidants as major components of plant defense operations. Considering recent informations gained through molecular-genetic studies, the current paper presents an overview of the structure, localization, biosynthesis (for GSH only), discusses GSH and GR significance in abiotic stress (such as salinity, drought, clilling, heavy metal)-exposed crop plants and also points out unexplored aspects in the current context for future studies.

Gill SS; Anjum NA; Hasanuzzaman M; Gill R; Trivedi DK; Ahmad I; Pereira E; Tuteja N

2013-09-01

322

Regeneration mechanisms of Arabidopsis thaliana methionine sulfoxide reductases B by glutaredoxins and thioredoxins.  

Science.gov (United States)

Methionine oxidation leads to the formation of S- and R-diastereomers of methionine sulfoxide (MetSO), which are reduced back to methionine by methionine sulfoxide reductases (MSRs) A and B, respectively. MSRBs are classified in two groups depending on the conservation of one or two redox-active Cys; 2-Cys MSRBs possess a catalytic Cys-reducing MetSO and a resolving Cys, allowing regeneration by thioredoxins. The second type, 1-Cys MSRBs, possess only the catalytic Cys. The biochemical mechanisms involved in activity regeneration of 1-Cys MSRBs remain largely elusive. In the present work we used recombinant plastidial Arabidopsis thaliana MSRB1 and MSRB2 as models for 1-Cys and 2-Cys MSRBs, respectively, to delineate the Trx- and glutaredoxin-dependent reduction mechanisms. Activity assays carried out using a series of cysteine mutants and various reductants combined with measurements of free thiols under distinct oxidation conditions and mass spectrometry experiments show that the 2-Cys MSRB2 is reduced by Trx through a dithiol-disulfide exchange involving both redox-active Cys of the two partners. Regarding 1-Cys MSRB1, oxidation of the enzyme after substrate reduction leads to the formation of a stable sulfenic acid on the catalytic Cys, which is subsequently glutathionylated. The deglutathionylation of MSRB1 is achieved by both mono- and dithiol glutaredoxins and involves only their N-terminal conserved catalytic Cys. This study proposes a detailed mechanism of the regeneration of 1-Cys MSRB activity by glutaredoxins, which likely constitute physiological reductants for this type of MSR. PMID:19457862

Tarrago, Lionel; Laugier, Edith; Zaffagnini, Mirko; Marchand, Christophe; Le Maréchal, Pierre; Rouhier, Nicolas; Lemaire, Stéphane D; Rey, Pascal

2009-05-20

323

Regeneration mechanisms of Arabidopsis thaliana methionine sulfoxide reductases B by glutaredoxins and thioredoxins.  

UK PubMed Central (United Kingdom)

Methionine oxidation leads to the formation of S- and R-diastereomers of methionine sulfoxide (MetSO), which are reduced back to methionine by methionine sulfoxide reductases (MSRs) A and B, respectively. MSRBs are classified in two groups depending on the conservation of one or two redox-active Cys; 2-Cys MSRBs possess a catalytic Cys-reducing MetSO and a resolving Cys, allowing regeneration by thioredoxins. The second type, 1-Cys MSRBs, possess only the catalytic Cys. The biochemical mechanisms involved in activity regeneration of 1-Cys MSRBs remain largely elusive. In the present work we used recombinant plastidial Arabidopsis thaliana MSRB1 and MSRB2 as models for 1-Cys and 2-Cys MSRBs, respectively, to delineate the Trx- and glutaredoxin-dependent reduction mechanisms. Activity assays carried out using a series of cysteine mutants and various reductants combined with measurements of free thiols under distinct oxidation conditions and mass spectrometry experiments show that the 2-Cys MSRB2 is reduced by Trx through a dithiol-disulfide exchange involving both redox-active Cys of the two partners. Regarding 1-Cys MSRB1, oxidation of the enzyme after substrate reduction leads to the formation of a stable sulfenic acid on the catalytic Cys, which is subsequently glutathionylated. The deglutathionylation of MSRB1 is achieved by both mono- and dithiol glutaredoxins and involves only their N-terminal conserved catalytic Cys. This study proposes a detailed mechanism of the regeneration of 1-Cys MSRB activity by glutaredoxins, which likely constitute physiological reductants for this type of MSR.

Tarrago L; Laugier E; Zaffagnini M; Marchand C; Le Maréchal P; Rouhier N; Lemaire SD; Rey P

2009-07-01

324

Thioredoxin reductase is irreversibly modified by curcumin: a novel molecular mechanism for its anticancer activity.  

Science.gov (United States)

The thioredoxin reductase (TrxR) isoenzymes, TrxR1 in cytosol or nucleus and TrxR2 in mitochondria, are essential mammalian selenocysteine (Sec)-containing flavoenzymes with a -Gly-Cys-Sec-Gly active site. TrxRs are the only enzymes catalyzing the NADPH-dependent reduction of the active site disulfide in thioredoxins (Trxs), which play essential roles in substrate reductions, defense against oxidative stress, and redox regulation by thiol redox control. TrxRs have been found to be overexpressed by a number of human tumors. Curcumin, which is consumed daily by millions of people, is a polyphenol derived from the plant Curcuma longa. This phytochemical has well known anticancer and antiangiogenic properties. In this study we report that rat TrxR1 activity in Trx-dependent disulfide reduction was inhibited by curcumin. The IC(50) value for the enzyme was 3.6 microM after incubation at room temperature for 2 h in vitro. The inhibition occurred with enzyme only in the presence of NADPH and persisted after removal of curcumin. By using mass spectrometry and blotting analysis, we proved that this irreversible inhibition by curcumin was caused by alkylation of both residues in the catalytically active site (Cys(496)/Sec(497)) of the enzyme. However, the curcumin-modified enzyme showed a strongly induced NADPH oxidase activity to produce reactive oxygen species. Inhibition of TrxR by curcumin added to cultured HeLa cells was also observed with an IC(50) of around 15 microM. Modification of TrxR by curcumin provides a possible mechanistic explanation for its cancer preventive activity, shifting the enzyme from an antioxidant to a prooxidant. PMID:15879598

Fang, Jianguo; Lu, Jun; Holmgren, Arne

2005-05-06

325

Thioredoxin reductase is irreversibly modified by curcumin: a novel molecular mechanism for its anticancer activity.  

UK PubMed Central (United Kingdom)

The thioredoxin reductase (TrxR) isoenzymes, TrxR1 in cytosol or nucleus and TrxR2 in mitochondria, are essential mammalian selenocysteine (Sec)-containing flavoenzymes with a -Gly-Cys-Sec-Gly active site. TrxRs are the only enzymes catalyzing the NADPH-dependent reduction of the active site disulfide in thioredoxins (Trxs), which play essential roles in substrate reductions, defense against oxidative stress, and redox regulation by thiol redox control. TrxRs have been found to be overexpressed by a number of human tumors. Curcumin, which is consumed daily by millions of people, is a polyphenol derived from the plant Curcuma longa. This phytochemical has well known anticancer and antiangiogenic properties. In this study we report that rat TrxR1 activity in Trx-dependent disulfide reduction was inhibited by curcumin. The IC(50) value for the enzyme was 3.6 microM after incubation at room temperature for 2 h in vitro. The inhibition occurred with enzyme only in the presence of NADPH and persisted after removal of curcumin. By using mass spectrometry and blotting analysis, we proved that this irreversible inhibition by curcumin was caused by alkylation of both residues in the catalytically active site (Cys(496)/Sec(497)) of the enzyme. However, the curcumin-modified enzyme showed a strongly induced NADPH oxidase activity to produce reactive oxygen species. Inhibition of TrxR by curcumin added to cultured HeLa cells was also observed with an IC(50) of around 15 microM. Modification of TrxR by curcumin provides a possible mechanistic explanation for its cancer preventive activity, shifting the enzyme from an antioxidant to a prooxidant.

Fang J; Lu J; Holmgren A

2005-07-01

326

Mechanism of action of clostridial glycine reductase: Isolation and characterization of a covalent acetyl enzyme intermediate  

Energy Technology Data Exchange (ETDEWEB)

Clostridial glycine reductase consists of proteins A, B, and C and catalyzes the reaction glycine + P{sub i} + 2e{sup {minus}} {yields} acetyl phosphate + NH{sub 4}{sup +}. Evidence was previously obtained that is consistent with the involvement of an acyl enzyme intermediate in this reaction. The authors now demonstrate that protein C catalyzes exchange of ({sup 32}P)P{sub i} into acetyl phosphate, providing additional support for an acetyl enzyme intermediate on protein C. Furthermore, they have isolated acetyl protein C and shown that it is qualitatively, catalytically competent. Acetyl protein C can be obtained through the forward reaction from protein C and Se-(carboxymethyl)selenocysteine-protein A, which is generated by the reaction of glycine with proteins A and B. Acetyl protein C can also be generated through the reverse reaction by the addition of acetyl phosphate to protein C. Both procedures lead to the same acetyl enzyme. The acetyl enzyme reacts with P{sub i} to give acetyl phosphate. When ({sup 14}C)acetyl protein C is denaturated with TCA and redissolved with urea, radioactivity remained associated with the protein. Treatment with KBH{sub 4} removes all the radioactivity associated with protein C, resulting in the formation of ({sup 14}C)ethanol. They conclude that a thiol group on protein C is acetylated. Proteins A and C together catalyze the exchange of tritium atoms from ({sup 3}H)H{sub 2}O into acetyl phosphate. This exchange reaction supports the proposal that an enol of the acetyl enzyme is an intermediate in the reaction sequence.

Arkowitz, R.A.; Abeles, R.H. (Brandeis Univ., Waltham, MA (USA))

1991-04-23

327

DNA-maleimide: an improved maleimide compound for electrophoresis-based titration of reactive thiols in a specific protein.  

UK PubMed Central (United Kingdom)

BACKGROUND: Thiol-mediated redox regulation of proteins plays a key role in many cellular processes. METHODS: To understand the redox status of cysteinyl thiol groups of the desired proteins, we developed a new maleimide reagent: a maleimide-conjugated single strand DNA, DNA-maleimide (DNA-Mal). RESULTS: DNA-Mal labelled proteins run as a distinct band on SDS-PAGE, with a discrete 9.32 kDa mobility shift per label regardless of the protein species or electrophoretic conditions. CONCLUSIONS: DNA-Mal labels free thiols like standard maleimide reagents, but possesses practical advantages in titration of the number and relative content of free thiols in a protein. GENERAL SIGNIFICANCE: The versatility of DNA molecule enhances the application of DNA-Mal in a broader range of cysteine containing proteins.

Hara S; Nojima T; Seio K; Yoshida M; Hisabori T

2013-04-01

328

Conjugate addition of thiols to p-benzoquinone monoketals. Attempts to prepare chiral synthetic equivalents of p-benzoquinone  

Energy Technology Data Exchange (ETDEWEB)

The conjugate addition of several thiols to achiral and chiral p-benzoquinone monoketals has been studied. A chiral synthetic equivalent of p-benzoquinone has been obtained in both enantiopure forms. (Author) 13 refs.

March, P. de; Escoda, M.; Figueredo, M.; Font, J.; Medrano, J. [Departamento de Quimica. Universitat Autonoma de Barcelon, Bellaterra, (Spain)

1997-09-01

329

Photochemical reactions of thiol-terminated self-assembled monolayers (SAMs) for micropatterning of gold nanoparticles and controlled surface functionality  

International Nuclear Information System (INIS)

This paper reported a facile method for the patterning of gold nanoparticles (AuNPs) on SiO2/Si by combining photochemical reaction and self-assembly techniques, and the conversion of surface functionality through thiol-ene click chemistry. The oxidation of terminal thiols in self-assembled monolayer of (3-mercaptopropyl)trimethoxysilane upon exposure to 254 nm UV light under ambient atmosphere was investigated. Chemically well-defined microstructures were obtained by UV irradiation through a mask, and subsequent immersion of the substrate into a dispersion of AuNPs resulted in site-specific assembly of AuNPs via Au-S covalent bond in the unexposed area. Thiol-ene “click” reaction between surface thiol-group and alkene-containing molecules under illumination of 365 nm UV light was also demonstrated. X-ray photoelectron spectroscopy study indicated the successful conversion of surface functionality.

1000-01-00

330

Photochemical reactions of thiol-terminated self-assembled monolayers (SAMs) for micropatterning of gold nanoparticles and controlled surface functionality  

Energy Technology Data Exchange (ETDEWEB)

This paper reported a facile method for the patterning of gold nanoparticles (AuNPs) on SiO{sub 2}/Si by combining photochemical reaction and self-assembly techniques, and the conversion of surface functionality through thiol-ene click chemistry. The oxidation of terminal thiols in self-assembled monolayer of (3-mercaptopropyl)trimethoxysilane upon exposure to 254 nm UV light under ambient atmosphere was investigated. Chemically well-defined microstructures were obtained by UV irradiation through a mask, and subsequent immersion of the substrate into a dispersion of AuNPs resulted in site-specific assembly of AuNPs via Au-S covalent bond in the unexposed area. Thiol-ene 'click' reaction between surface thiol-group and alkene-containing molecules under illumination of 365 nm UV light was also demonstrated. X-ray photoelectron spectroscopy study indicated the successful conversion of surface functionality.

Han Xuemingyue; Wu Chong [National Center for Nanoscience and Technology, 11 Beiyitiao, Zhongguancun, Beijing 100190 (China); Graduate School of Chinese Academy of Sciences, Beijing 100049 (China); Sun Shuqing, E-mail: sun.shuqing@sz.tsinghua.edu.cn [Laboratory of Optical Imaging and Sensing, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055 (China)

2012-04-01

331

Photochemical reactions of thiol-terminated self-assembled monolayers (SAMs) for micropatterning of gold nanoparticles and controlled surface functionality  

Science.gov (United States)

This paper reported a facile method for the patterning of gold nanoparticles (AuNPs) on SiO2/Si by combining photochemical reaction and self-assembly techniques, and the conversion of surface functionality through thiol-ene click chemistry. The oxidation of terminal thiols in self-assembled monolayer of (3-mercaptopropyl)trimethoxysilane upon exposure to 254 nm UV light under ambient atmosphere was investigated. Chemically well-defined microstructures were obtained by UV irradiation through a mask, and subsequent immersion of the substrate into a dispersion of AuNPs resulted in site-specific assembly of AuNPs via Au-S covalent bond in the unexposed area. Thiol-ene "click" reaction between surface thiol-group and alkene-containing molecules under illumination of 365 nm UV light was also demonstrated. X-ray photoelectron spectroscopy study indicated the successful conversion of surface functionality.

Han, Xuemingyue; Wu, Chong; Sun, Shuqing

2012-04-01

332

Native chemical ligation,thiol-ene click: a methodology for the synthesis of functionalized peptides.  

Science.gov (United States)

The sequential combination of native chemical ligation and thiol-ene radical chemistry (NCL-TEC) on the resulting cysteine thiol has been investigated as a methodology for rapidly accessing functionalized peptides. Three sequential cycles of native chemical ligation and subsequent thiyl radical reactions (including a free-radical-mediated desulfurization reaction) were carried out on a peptide backbone demonstrating the iterative nature of this process. The versatility of the thiyl radical reaction at cysteine was demonstrated through the introduction of a number of different side chains including an amino acid derivative, a carbohydrate group, and an alkyl azide. Conditions were developed that allowed the sequential NCL-TEC process to proceed in high yield. PMID:23565861

Markey, Lyn; Giordani, Silvia; Scanlan, Eoin M

2013-04-18

333

Native chemical ligation,thiol-ene click: a methodology for the synthesis of functionalized peptides.  

UK PubMed Central (United Kingdom)

The sequential combination of native chemical ligation and thiol-ene radical chemistry (NCL-TEC) on the resulting cysteine thiol has been investigated as a methodology for rapidly accessing functionalized peptides. Three sequential cycles of native chemical ligation and subsequent thiyl radical reactions (including a free-radical-mediated desulfurization reaction) were carried out on a peptide backbone demonstrating the iterative nature of this process. The versatility of the thiyl radical reaction at cysteine was demonstrated through the introduction of a number of different side chains including an amino acid derivative, a carbohydrate group, and an alkyl azide. Conditions were developed that allowed the sequential NCL-TEC process to proceed in high yield.

Markey L; Giordani S; Scanlan EM

2013-05-01

334

Thiol- and Biotin-Labeled Probes for Oligonucleotide Quartz Crystal Microbalance Biosensors of Microalga Alexandrium Minutum  

Directory of Open Access Journals (Sweden)

Full Text Available Two quartz crystal microbalance oligonucleotide biosensors of a toxic microalga gene sequence (Alexandrium Minutum) have been designed. Grafting on a gold surface of 20-base thiol- or biotin-labeled probe, and selective hybridization with the complementary 20-base target, have been monitored in situ with a 27 MHz quartz crystal microbalance under controlled hydrodynamic conditions. The frequency of the set up is stable to within a few hertz, corresponding to the nanogram scale, for three hour experiments. DNA recognition by the two biosensors is efficient and selective. Hybridization kinetic curves indicate that the biosensor designed with the thiol-labeled probe is more sensitive, and that the biosensor designed with the biotin-labeled probe has a shorter time response and a higher hybridization efficiency.

Mathieu Lazerges; Hubert Perrot; Niriniony Rabehagasoa; Chantal Compère

2012-01-01

335

Fabrication of gold micro- and nanostructures by photolithographic exposure of thiol-stabilized gold nanoparticles.  

UK PubMed Central (United Kingdom)

Exposure of thiol-stabilized gold nanoparticles supported on silicon wafers to UV light leads to oxidation of the thiol molecules and coagulation of the nanoparticles, forming densified structures that are resistant to removal by solvent exposure. Unoxidized particles may, in contrast, readily be removed leaving gold structures behind at the surface. This process provides a convenient and simple route for the fabrication of gold structures with dimensions ranging from micrometers to nanometers. The use of masks enables micrometer-scale structures to be fabricated rapidly. Exposure of nanoparticles to light from a near-field scanning optical microscope (NSOM) leads to the formation of gold nanowires. The dimensions of these nanowires depend on the method of preparation of the film: for spin-cast films, a width of 200 nm was achieved. However, this was reduced significantly, to 60 nm, for Langmuir-Schaeffer films.

Sun S; Mendes P; Critchley K; Diegoli S; Hanwell M; Evans SD; Leggett GJ; Preece JA; Richardson TH

2006-03-01

336

Fabrication of gold micro- and nanostructures by photolithographic exposure of thiol-stabilized gold nanoparticles.  

Science.gov (United States)

Exposure of thiol-stabilized gold nanoparticles supported on silicon wafers to UV light leads to oxidation of the thiol molecules and coagulation of the nanoparticles, forming densified structures that are resistant to removal by solvent exposure. Unoxidized particles may, in contrast, readily be removed leaving gold structures behind at the surface. This process provides a convenient and simple route for the fabrication of gold structures with dimensions ranging from micrometers to nanometers. The use of masks enables micrometer-scale structures to be fabricated rapidly. Exposure of nanoparticles to light from a near-field scanning optical microscope (NSOM) leads to the formation of gold nanowires. The dimensions of these nanowires depend on the method of preparation of the film: for spin-cast films, a width of 200 nm was achieved. However, this was reduced significantly, to 60 nm, for Langmuir-Schaeffer films. PMID:16522020

Sun, Shuqing; Mendes, Paula; Critchley, Kevin; Diegoli, Sara; Hanwell, Marcus; Evans, Stephen D; Leggett, Graham J; Preece, Jon A; Richardson, Tim H

2006-03-01

337

XAFS, SAXS and HREM characterization of Pd nanoparticles capped with n-alkyl thiol molecules  

International Nuclear Information System (INIS)

We report a structural characterization of palladium nanoparticles, with an average diameter of 1.2 nm, capped with three different n-alkyl thiol molecules (n=12, 16 and 18). For this purpose several experimental techniques were used, namely X-ray absorption fine structure (XAFS), small angle X-ray scattering (SAXS) and high-resolution electron transmission microscopy (HRTEM). SAXS and HRTEM results yielded the sizes of the Pd nanoparticles in each sample. The effects of sulfur-palladium interaction on the structural and electronic properties of the studied material were derived from XAFS results. This study indicates the presence of sulfurized palladium in the bulk of the three studied capped Pd nanoparticles. Slight variations in the degree of sulfidation were detected for nanoclusters with different thiol chain lengths.

2007-02-01

338

High-performance liquid chromatographic separation and indirect fluorescence detection of thiols.  

Science.gov (United States)

A fluorescent post-column reaction detection scheme has been devised for selective determination of thiols. The post-column reagent is 40 microM Cd2+ and 100 microM 8-hydroxyquinoline-5-sulfonic acid (HQS) in non-complexing buffer at pH 10. HQS complexes Cd2+ to form a fluorescent product. Thiols in the HPLC effluent compete for complexation of the Cd2+, resulting in a decrease in the fluorescence response. Detection limits of 0.2 microM (0.04 ppm) are achieved for cysteine, homocysteine and glutathione in a 5 min separation. Recoveries from spiked synthetic urine samples are 87-120%. PMID:12416880

Pelletier, Sarah; Lucy, Charles A

2002-10-01

339

Pendant thiol groups-attached Pd(II) for initiating metal deposition  

Energy Technology Data Exchange (ETDEWEB)

A new activation method has been developed for initiating electroless metal deposition on silicon substrates without SnCl{sub 2} sensitization and roughening condition. Silicon wafers are first coated with thiol-terminated self-assembled monolayers (SAMs), and then catalyzed with a stable tin-free Pd(II)-based colloidal solution. Atomic force microscopy (AFM), Auger electron spectroscopy (AES) and X-ray photoelectron spectroscopy (XPS) were used to characterize the step-by-step surfaces and study the binding mechanism of Pd(II) with SAMs onto surfaces. Results show that Pd(II) oligomer particles are chemisorbed on pendant thiol surfaces through S-Pd bonds. This process involves fewer steps than the conventional Sn/Pd combined activation one. Furthermore, the chemical bound initiator possesses longevity and can be stored for a long time before metallization.

Xu Lina; Liao Jianhui; Huang Lan; Gu Ning; Zhang Haiqian; Liu Juzheng

2003-04-30

340

Protection with combinations of hydroxytryptophan and some thiol compounds against whole-body gamma irradiation  

Energy Technology Data Exchange (ETDEWEB)

Protection of mice (as measured by survival at 30 days) against whole-body gamma exposure by prior treatment with combinations of hydroxytryptophan (HT) and some thiol drugs, is reported here. The combination of HT and aminoethylisothiuronium bromide hydrobromide (AET) was much more effective than HT alone against the effects of 10.5 and 12.5 Gy of gamma ray exposure. A combination of HT and ..beta..-mercaptopropionylglycine (MPG) gave marked protection against a 10.5 Gy exposure but not against 12.5 Gy. The combination of HT with cysteine or cysteamine also provided reasonable protection against 10.5 Gy. The combination of HT and cystamine rendered a low level of protection against 10.5 Gy, but again it was better than HT alone. The thiol drugs by themselves in the doses used in combination were ineffective in rendering protection after whole body gamma ray exposure.

Ghose, A.; Ganguly, S.K.; Kaur, J. (Institute of Nuclear Medicine and Allied Sciences, Delhi (India))

1983-08-01

 
 
 
 
341

Investigation of ethyl radical quenching by phenolics and thiols in model wine.  

UK PubMed Central (United Kingdom)

In the present study, the reaction between 1-hydroxyethyl radicals (1-HER) and various wine-related phenolics and thiols, including gallic acid, caffeic acid, ferulic acid, 3-mercaptohexan-1-ol (3MH), cysteine (Cys), and glutathione (GSH), was studied using competitive spin trapping with electron paramagnetic resonance (EPR) and mass spectrometry. Previous studies have reported several important reactions occurring between quinones and other wine components, but the fate of 1-HER within the context of wine oxidation is less understood. Furthermore, the ability of these compounds to prevent formation of acetaldehyde, a known nonenzymatic oxidation product of ethanol, was measured. The hydroxycinnamic acids and thiol compounds tested at 5 mM concentrations significantly inhibited spin adduct formation, indicating their reactivity toward 1-HER. In addition, we confirm that loss of 3MH under model wine conditions is due to quinone trapping as well as 1-HER-induced oxidation.

Kreitman GY; Laurie VF; Elias RJ

2013-01-01

342

Bridged nucleic acid conjugates at 6'-thiol: synthesis, hybridization properties and nuclease resistances.  

Science.gov (United States)

The bridged nucleic acid (BNA) containing a thiol at the 6'-position in the bridged structure was synthesized from the disulfide-type BNA and conjugated with various functional molecules via the thioether or the disulfide linkage post-synthetically and efficiently in solution phase. The disulfide-linked conjugate was cleaved under reductive conditions derived from glutathione and an oligonucleotide bearing a free thiol was released smoothly. Conjugated functional molecules had great effects on duplex stability with the DNA complement. In contrast, the molecules little influenced the stability with the RNA complement. Moreover, the oligonucleotides with functional groups at the 6'-position had as high or higher resistances against 3'-exonuclease than phosphorothioate oligonucleotide (S-oligo). PMID:21643564

Mori, Kazuto; Kodama, Tetsuya; Baba, Takeshi; Obika, Satoshi

2011-06-03

343

Bridged nucleic acid conjugates at 6'-thiol: synthesis, hybridization properties and nuclease resistances.  

UK PubMed Central (United Kingdom)

The bridged nucleic acid (BNA) containing a thiol at the 6'-position in the bridged structure was synthesized from the disulfide-type BNA and conjugated with various functional molecules via the thioether or the disulfide linkage post-synthetically and efficiently in solution phase. The disulfide-linked conjugate was cleaved under reductive conditions derived from glutathione and an oligonucleotide bearing a free thiol was released smoothly. Conjugated functional molecules had great effects on duplex stability with the DNA complement. In contrast, the molecules little influenced the stability with the RNA complement. Moreover, the oligonucleotides with functional groups at the 6'-position had as high or higher resistances against 3'-exonuclease than phosphorothioate oligonucleotide (S-oligo).

Mori K; Kodama T; Baba T; Obika S

2011-07-01

344

The Role of Thiol on Degradation of Pentaerythrityl Tetranitrate and Isosorbide Dinitrate  

Directory of Open Access Journals (Sweden)

Full Text Available Thiols such as N-acetylcystein (NAC) are used to replenish glutathione (GSH) level, with regard to their function in the maintenance of cellular reduction-oxidation balance and control of oxidative stress. Thiols play a role in the reductive metabolism of nitrates to NO, an important signaling molecule in the cardiovascular system as well as other systems throughout the body. This study aimed to evaluate the influence of NAC on decomposition of different organic nitrate esters according to its potential i.e., pentaerythrityl tetranitrate (PETN) and isosorbide dinitrate (ISDN). The results showed that NAC gives a rapid and significant decrease of PETN and ISDN during the incubation period. During the experiment, about 85% of PETN were decomposed, while the decomposition of ISDN was about 20%. Detection of nitrite and elucidation of disulphide bond of NAC gives evidence that confirms the presence of reactions.

M.R. Suwitono; R.E. Kartasasmita; J.S. Pamudji; S. Ibrahim

2011-01-01

345

Occurrence of odorant polyfunctional thiols in the Super Alpha Tomahawk hop cultivar. Comparison with the thiol-rich Nelson Sauvin bitter variety.  

UK PubMed Central (United Kingdom)

Tomahawk hop (Humulus lupulus) is a recently developed Super Alpha cultivar (14-18% ?-acids w/w), already widely used by brewers to impart bitterness and a citrus-like aroma to beer. By comparison with two bitter varieties (Nelson Sauvin and Nugget) and two aromatic ones (Cascade and Saaz), the Tomahawk cultivar showed a very particular terpenoid profile, rich in both ?- and ?-selinenes (>600 mg/kg IST equiv in total), methyl geranate (>40 mg/kg IST equiv), and geraniol (>200 mg/kg). Tomahawk also proved to contain a wide variety of odorant polyfunctional thiols. The major ?-sulfanyl acetate, 3-sulfanyl-2-ethylpropyl acetate, newly identified here, was found at similar levels in the famous Sauvignon-like Nelson Sauvin and Tomahawk varieties (15-44 ?g/kg IST equiv). On the other hand, lower levels of total ?-sulfanyl alcohols were measured in Tomahawk, although 3-sulfanylhexan-1-ol was found at a similar level and the 3-sulfanyl-4-methylpentan-1-ol previously claimed to be specific to the Nelson Sauvin variety was also evidenced in the Super Alpha cultivar (9-13 ?g/kg IST equiv). As revealed by boiling and fermentation, Tomahawk hop also contains very interesting bound polyfunctional thiols that should be investigated for better use by brewers.

Gros J; Nizet S; Collin S

2011-08-01

346

Characterization of mitochondrial thioredoxin reductase from C. elegans  

International Nuclear Information System (INIS)

Thioredoxin reductase catalyzes the NADPH-dependent reduction of the catalytic disulfide bond of thioredoxin. In mammals and other higher eukaryotes, thioredoxin reductases contain the rare amino acid selenocysteine at the active site. The mitochondrial enzyme from Caenorhabditis elegans, however, contains a cysteine residue in place of selenocysteine. The mitochondrial C. elegans thioredoxin reductase was cloned from an expressed sequence tag and then produced in Escherichia coli as an intein-fusion protein. The purified recombinant enzyme has a k cat of 610 min-1 and a K m of 610 ?M using E. coli thioredoxin as substrate. The reported k cat is 25% of the k cat of the mammalian enzyme and is 43-fold higher than a cysteine mutant of mammalian thioredoxin reductase. The enzyme would reduce selenocysteine, but not hydrogen peroxide or insulin. The flanking glycine residues of the GCCG motif were mutated to serine. The mutants improved substrate binding, but decreased the catalytic rate

2006-08-04

347

Betaine for treatment of homocystinuria caused by methylenetetrahydrofolate reductase deficiency.  

Science.gov (United States)

A 24 day old girl with homocystinuria and hypomethioninaemia caused by methylenetetrahydrofolate reductase deficiency presented with rapidly progressing encephalopathy and myopathy. An almost complete recovery was achieved by treatment with betaine.

Holme, E; Kjellman, B; Ronge, E

1989-01-01

348

A study of oxidative stress, thiol proteins and role of vitamin E supplementation in chronic obstructive pulmonary disease (COPD)  

Directory of Open Access Journals (Sweden)

Full Text Available Background: Lipid peroxide plays an important role in inflammatory lung disease. Increased epithelial permeability produced by cigarette smoke is likely to be mediated through depletion of thiol proteins. Imbalance between oxidants and thiol proteins is also an established fact in these patients. Materials & methods: In the present study 30 healthy non-smokers were served as controls and 20 patients with stable COPD were included. Their base line clinical examination, Malondialdehyde (MDA) as an oxidant, alpha tocopherol and erythrocyte superoxide dismutase (SOD) as an antioxidants and thiol proteins levels were measured. All above parameters were repeated after 12 weeks of supplementation with 400 IU of vitamin E daily. Results: We observed that the mean malondialdehyde levels in these patients at base line were high (p<0.001) than Control Plasma alpha-tocopherol, SOD and thiol proteins levels were low (p<0.001) in the patients compared to controls. Exogenous vitamin E (400 IU twice daily) Supplementation did not bring about any significant change in plasma Erythrocyte Superoxide Dismutase and vitamin E. But slight increase in the plasma thiol proteins levels was seen. The present study shows that initially the plasma lipid peroxide (MDA) levels were high antioxidant (alpha- tocopherol, SOD) and thiol proteins were low in patients with COPD. Exogenous supplementation with vitamin E increases slightly thiol proteins levels and brings down the levels of MDA showing attenuation of further damage. Conclusion: Our study confirmed the existence of oxidative stress and and the augmentation of antioxidant defenses as shown by slight increase in thiol proteins level. The antioxidant therapy is adjunct in lung disease patients and opens a promising field in prevention of oxidative stress related complications in these patients.

Anita M. Raut; Adinath N. Suryakar; Dilip Mhaisekar

2013-01-01

349

Characterization of N-ethylmaleimide-sensitive thiol groups required for the GTP-dependent fusion of endoplasmic reticulum membranes.  

Science.gov (United States)

The GTP-dependent fusion activity of endoplasmic reticulum membranes is thought to be required for the structural maintenance and post-mitotic regeneration of the endoplasmic reticulum. This fusion is sensitive to the thiol-alkylating agent N-ethylmaleimide. In many intracellular fusion events N-ethylmaleimide-sensitivity is associated with a homotrimeric ATPase called N-ethylmaleimide-sensitive fusion protein or NSF. The addition of cytosol containing NSF is known to restore fusion activity to N-ethylmaleimide-treated membranes. We found that the inhibition of fusion of rat liver endoplasmic reticulum membranes (microsomes) by N-ethylmaleimide was not reversed by the addition of untreated cytosol. Fusion was also unaffected by treatment with a buffer known to remove NSF from membranes. Accordingly, no membrane-associated NSF was detected by immunoblot analysis. These data suggest that microsome fusion requires an N-ethylmaleimide-sensitive component distinct from NSF. This component was tightly associated with the membranes, so we used a number of chemical probes to characterize it in situ. Its thiol groups did not appear to be part of a GTP-binding site. They showed relatively low reactivity with sodium periodate, which induces the formation of disulphide bonds between proximate thiol groups. The thiols were not protected against N-ethylmaleimide by Zn2+, a potent inhibitor of fusion which is known to efficiently co-ordinate thiol groups. To characterize the topology of the fusion-related thiol groups we used bulky thiol-specific reagents prepared by conjugating BSA or 10 kDa aminodextran to the bifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. The inhibition of fusion by these reagents indicated that these thiols are highly exposed on the membranes. This exposure might be important for the function of these groups during GTP-triggered fusion. PMID:7492317

Sokoloff, A V; Whalley, T; Zimmerberg, J

1995-11-15

350

Characterization of N-ethylmaleimide-sensitive thiol groups required for the GTP-dependent fusion of endoplasmic reticulum membranes.  

UK PubMed Central (United Kingdom)

The GTP-dependent fusion activity of endoplasmic reticulum membranes is thought to be required for the structural maintenance and post-mitotic regeneration of the endoplasmic reticulum. This fusion is sensitive to the thiol-alkylating agent N-ethylmaleimide. In many intracellular fusion events N-ethylmaleimide-sensitivity is associated with a homotrimeric ATPase called N-ethylmaleimide-sensitive fusion protein or NSF. The addition of cytosol containing NSF is known to restore fusion activity to N-ethylmaleimide-treated membranes. We found that the inhibition of fusion of rat liver endoplasmic reticulum membranes (microsomes) by N-ethylmaleimide was not reversed by the addition of untreated cytosol. Fusion was also unaffected by treatment with a buffer known to remove NSF from membranes. Accordingly, no membrane-associated NSF was detected by immunoblot analysis. These data suggest that microsome fusion requires an N-ethylmaleimide-sensitive component distinct from NSF. This component was tightly associated with the membranes, so we used a number of chemical probes to characterize it in situ. Its thiol groups did not appear to be part of a GTP-binding site. They showed relatively low reactivity with sodium periodate, which induces the formation of disulphide bonds between proximate thiol groups. The thiols were not protected against N-ethylmaleimide by Zn2+, a potent inhibitor of fusion which is known to efficiently co-ordinate thiol groups. To characterize the topology of the fusion-related thiol groups we used bulky thiol-specific reagents prepared by conjugating BSA or 10 kDa aminodextran to the bifunctional reagent N-succinimidyl 3-(2-pyridyldithio)propionate. The inhibition of fusion by these reagents indicated that these thiols are highly exposed on the membranes. This exposure might be important for the function of these groups during GTP-triggered fusion.

Sokoloff AV; Whalley T; Zimmerberg J

1995-11-01

351

A convenient preparation of heteroaryl sulfonamides and sulfonyl fluorides from heteroaryl thiols.  

UK PubMed Central (United Kingdom)

Heteroaromatic thiols may be oxidized to the sulfonyl chloride at low temperature (-25 degrees C) by using 3.3 equiv of aqueous sodium hypochlorite. The reaction is rapid, avoids the use of chlorine gas, and succeeds with substrates that have previously been found to afford little or none of the sulfonamide product with other procedures. The method allows the preparation of the sulfonyl fluorides, which are stable enough to be purified and stored, making them potentially useful monomers in parallel chemistry efforts.

Wright SW; Hallstrom KN

2006-02-01

352

Sc(III)-catalyzed enantioselective addition of thiols to ?,?-unsaturated ketones in neutral water.  

UK PubMed Central (United Kingdom)

This report concerns Lewis acid catalyzed enantioselective sulfa-Michael addition in neutral water by using a very efficient Sc(OTf)(3)/bipyridine 1 catalytic system. It is noteworthy that the protocol presented employs water as a reaction medium and allows us to obtain very high stereoselectivity and satisfactory yields for ?-keto sulphides deriving from aliphatic thiols. The recovery and reuse of both the aqueous medium and the catalytic system is also reported.

Bonollo S; Lanari D; Pizzo F; Vaccaro L

2011-05-01

353

Electrophoretic artifacts arising from the use of thiol-containing reagents.  

UK PubMed Central (United Kingdom)

Thiol reagents migrate as a curtain behind the salt front when loaded with the sample solution onto disc-electrophoresis gels. In immobilized pH gradients (IPG) the same compounds are driven by electrophoresis and electroosmosis from the alkaline to the neutral and acidic regions of the gradients. In either case, a dose-dependent sideways spreading results in spurious reduction between adjacent lanes if samples with and without reducing agent are loaded side by side.

Gianazza E; De Ponti P

1993-12-01

354

Thiol dosing of ZnO single crystals and nanorods: Surface chemistry and photoluminescence  

Science.gov (United States)

Adsorption of thiols on ZnO(0001) and ZnO nanorods has been investigated using X-ray and ultraviolet photoelectron spectroscopies (XPS and UPS). Ultrahigh vacuum (UHV) dosing of sputter-cleaned ZnO(0001) with methanethiol (MT), 1-dodecanethiol (DDT), and 3-mercaptopropyltrimethoxysilane (MPTMS) leads to S2p peaks with a binding energy of 163.3 eV. Similar results for MPTMS are obtained for sputter-cleaned ZnO(0001) that is pre-dosed with water to form hydroxyl groups. In all cases, the absence of a free thiol S2p peak at 164.2 eV indicates that bonding to the surface occurs via the thiol end of the molecule. A DDT-dosed ZnO(0001) sample stored for 10 days in UHV and heated to temperatures as high as 150 °C exhibits minimal changes in its S/Zn atomic ratio, confirming chemisorption and the presence of a strong bond to the surface. UPS shows that MT adsorption on sputtered ZnO(0001) leads to a 0.7 eV increase in work function and perturbation of the MT molecular orbitals, again consistent with chemisorption. Dry ZnO nanorods have been exposed to MT while monitoring their photoluminescence. XPS and Raman spectroscopy confirm thiol adsorption. Relative to dry ZnO, adsorption causes a decrease in intensity of the visible emission peak, but the UV peak remains unchanged. These results indicate that ZnS bond formation quenches radiative decay to the valence band from defect states, possibly by methanethiolate adsorption filling oxygen vacancies.

Singh, Jagdeep; Im, Jisun; Watters, Evan J.; Whitten, James E.; Soares, Jason W.; Steeves, Diane M.

2013-03-01

355

Ruthenium(III) Chloride Catalyzed Acylation of Alcohols, Phenols, and Thiols in Room Temperature Ionic Liquids  

Directory of Open Access Journals (Sweden)

Full Text Available Ruthenium(III) chloride-catalyzed acylation of a variety of alcohols, phenols, and thiols was achieved in high yields under mild conditions (room temperature) in the ionic liquid 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]). The ionic liquid and ruthenium catalyst can be recycled at least 10 times. Our system not only solves the basic problem of ruthenium catalyst reuse, but also avoids the use of volatile acetonitrile as solvent.

Zhiwen Xi; Wenyan Hao; Pingping Wang; Mingzhong Cai

2009-01-01

356

Aldose reductase inhibitors from the fruits of Caesalpinia ferrea Mart.  

UK PubMed Central (United Kingdom)

Aldose reductase inhibitors were isolated from an extract of the dry fruits of Caesalpinia ferrea Mart. (Leguminosae). Compound 2 was identified as ellagic acid by comparison with a reference sample. The structure of compound 1 was elucidated as 2-(2,3,6-trihydroxy-4-carboxyphenyl) ellagic acid on the basis of spectral evidence, especially 2D-NMR data (HMQC, HMBC and NOESY). These two compounds inhibited aldose reductase in a non-competitive manner.

Ueda H; Tachibana Y; Moriyasu M; Kawanishi K; Alves SM

2001-09-01

357

Aldose reductase inhibitors from the fruits of Caesalpinia ferrea Mart.  

Science.gov (United States)

Aldose reductase inhibitors were isolated from an extract of the dry fruits of Caesalpinia ferrea Mart. (Leguminosae). Compound 2 was identified as ellagic acid by comparison with a reference sample. The structure of compound 1 was elucidated as 2-(2,3,6-trihydroxy-4-carboxyphenyl) ellagic acid on the basis of spectral evidence, especially 2D-NMR data (HMQC, HMBC and NOESY). These two compounds inhibited aldose reductase in a non-competitive manner. PMID:11695881

Ueda, H; Tachibana, Y; Moriyasu, M; Kawanishi, K; Alves, S M

2001-09-01

358

Glutathione peroxidases and glutathione reductase activities during Bufo bufo development.  

UK PubMed Central (United Kingdom)

Glutathione peroxidases and glutathione reductase activities are expressed from the early stage of Bufo bufo development. Selenium-dependent and selenium-independent glutathione peroxidase activities fluctuated independently. The activity of selenium-independent was found to be higher than that of selenium-dependent glutathione peroxidase through all stages of development. Glutathione reductase activity, after a slight fall from stage 4 to stage 7, constantly increased up to stage 25.

Di Ilio C; Del Boccio G; Miranda M; Manilla A; Zarivi O; Federici G

1986-01-01

359

Glutathione peroxidases and glutathione reductase activities during Bufo bufo development.  

Science.gov (United States)

Glutathione peroxidases and glutathione reductase activities are expressed from the early stage of Bufo bufo development. Selenium-dependent and selenium-independent glutathione peroxidase activities fluctuated independently. The activity of selenium-independent was found to be higher than that of selenium-dependent glutathione peroxidase through all stages of development. Glutathione reductase activity, after a slight fall from stage 4 to stage 7, constantly increased up to stage 25. PMID:2869913

Di Ilio, C; Del Boccio, G; Miranda, M; Manilla, A; Zarivi, O; Federici, G

1986-01-01

360

The interplay between thiol-compounds against chromium (VI) in the freshwater green alga Monoraphidium convolutum: toxicology, photosynthesis, and oxidative stress at a glance.  

Science.gov (United States)

In this paper, the multifaceted Cr(VI) toxicity over the freshwater green alga Monoraphidium convolutum was assessed by concomitantly monitoring thiol-dependent redox balances, photosynthesis activity and growth-survival scores. Control group showed exponential growth rate at (5.78±0.29) division/day until 8th day with linear increasing chlorophyll a/protein ratios (CHLa/PROT) throughout the period. Cultures of M. convolutum were exposed for 5 days to Cr(VI) concentrations from 0 up to 100mg/L showing that CHLa/PROT ratios were sensibly affected, in agreement to the calculated LC(50,48 h) (5.38±0.72) mg/L from the concentration-response curve of cell mortality after 48 h. Regarding photosynthesis effects, Cr(VI) concentrations >1.0 mg/L showed significant increases in short-term (after 2 h) electron transfer rates (ETR) and quantum yields of photosystem II (?(PSII)), followed by subsequent decline of both parameters after 48 and 72 h. Biochemical analyses showed that maximal GSH concentrations in algal cultures were observed upon 1mg Cr(VI)/L and higher dichromate concentrations dramatically increased the activity of antioxidant GSH-dependent enzymes ascorbate peroxidase and glutathione reductase. However, no variation was observed in the cellular GSH levels, whereas GSSG and lipid peroxidation indexes abruptly increased upon 10 mg Cr(VI)/L exposure. Altogether, plant physiology, photosynthesis and biochemical data suggest that the GSH-dependent antioxidant system is capable to sustain M. convolutum viability through efficient photosynthesis activity and adequate antioxidant responses up to Cr(VI) concentrations of 1.0mg/L, when redox unbalances were first evidenced. PMID:22522782

Takami, R; Almeida, J V; Vardaris, C V; Colepicolo, P; Barros, M P

2012-04-03

 
 
 
 
361

The interplay between thiol-compounds against chromium (VI) in the freshwater green alga Monoraphidium convolutum: Toxicology, photosynthesis, and oxidative stress at a glance  

Energy Technology Data Exchange (ETDEWEB)

In this paper, the multifaceted Cr(VI) toxicity over the freshwater green alga Monoraphidium convolutum was assessed by concomitantly monitoring thiol-dependent redox balances, photosynthesis activity and growth-survival scores. Control group showed exponential growth rate at (5.78 {+-} 0.29) division/day until 8th day with linear increasing chlorophyll a/protein ratios (CHLa/PROT) throughout the period. Cultures of M. convolutum were exposed for 5 days to Cr(VI) concentrations from 0 up to 100 mg/L showing that CHLa/PROT ratios were sensibly affected, in agreement to the calculated LC{sub 50,48h} (5.38 {+-} 0.72) mg/L from the concentration-response curve of cell mortality after 48 h. Regarding photosynthesis effects, Cr(VI) concentrations >1.0 mg/L showed significant increases in short-term (after 2 h) electron transfer rates (ETR) and quantum yields of photosystem II ({Phi}{sub PSII}), followed by subsequent decline of both parameters after 48 and 72 h. Biochemical analyses showed that maximal GSH concentrations in algal cultures were observed upon 1 mg Cr(VI)/L and higher dichromate concentrations dramatically increased the activity of antioxidant GSH-dependent enzymes ascorbate peroxidase and glutathione reductase. However, no variation was observed in the cellular GSH levels, whereas GSSG and lipid peroxidation indexes abruptly increased upon 10 mg Cr(VI)/L exposure. Altogether, plant physiology, photosynthesis and biochemical data suggest that the GSH-dependent antioxidant system is capable to sustain M. convolutum viability through efficient photosynthesis activity and adequate antioxidant responses up to Cr(VI) concentrations of 1.0 mg/L, when redox unbalances were first evidenced.

Takami, R. [Postgraduate Program in Environmental Chemistry, CBS, Universidade Cruzeiro do Sul, 08060070, Sao Paulo, SP (Brazil); Almeida, J.V. [Department of Biochemistry, Instituto de Quimica, Universidade de Sao Paulo (IQ-USP), Sao Paulo, SP (Brazil); Vardaris, C.V. [Postgraduate Program in Environmental Chemistry, CBS, Universidade Cruzeiro do Sul, 08060070, Sao Paulo, SP (Brazil); Colepicolo, P. [Department of Biochemistry, Instituto de Quimica, Universidade de Sao Paulo (IQ-USP), Sao Paulo, SP (Brazil); Barros, M.P., E-mail: marcelo.barros@cruzeirodosul.edu.br [Postgraduate Program in Environmental Chemistry, CBS, Universidade Cruzeiro do Sul, 08060070, Sao Paulo, SP (Brazil)

2012-08-15

362

The interplay between thiol-compounds against chromium (VI) in the freshwater green alga Monoraphidium convolutum: Toxicology, photosynthesis, and oxidative stress at a glance  

International Nuclear Information System (INIS)

In this paper, the multifaceted Cr(VI) toxicity over the freshwater green alga Monoraphidium convolutum was assessed by concomitantly monitoring thiol-dependent redox balances, photosynthesis activity and growth-survival scores. Control group showed exponential growth rate at (5.78 ± 0.29) division/day until 8th day with linear increasing chlorophyll a/protein ratios (CHLa/PROT) throughout the period. Cultures of M. convolutum were exposed for 5 days to Cr(VI) concentrations from 0 up to 100 mg/L showing that CHLa/PROT ratios were sensibly affected, in agreement to the calculated LC50,48h (5.38 ± 0.72) mg/L from the concentration-response curve of cell mortality after 48 h. Regarding photosynthesis effects, Cr(VI) concentrations >1.0 mg/L showed significant increases in short-term (after 2 h) electron transfer rates (ETR) and quantum yields of photosystem II (?PSII), followed by subsequent decline of both parameters after 48 and 72 h. Biochemical analyses showed that maximal GSH concentrations in algal cultures were observed upon 1 mg Cr(VI)/L and higher dichromate concentrations dramatically increased the activity of antioxidant GSH-dependent enzymes ascorbate peroxidase and glutathione reductase. However, no variation was observed in the cellular GSH levels, whereas GSSG and lipid peroxidation indexes abruptly increased upon 10 mg Cr(VI)/L exposure. Altogether, plant physiology, photosynthesis and biochemical data suggest that the GSH-dependent antioxidant system is capable to sustain M. convolutum viability through efficient photosynthesis activity and adequate antioxidant responses up to Cr(VI) concentrations of 1.0 mg/L, when redox unbalances were first evidenced.

2012-08-15

363

Surface modification of CdS quantum dots using thiols-structural and photophysical studies  

International Nuclear Information System (INIS)

This study is aimed at identifying a suitable organic thiol for CdS by studying its structural, thermal and photophysical characteristics. Quantum dots of the II-VI semiconductor CdS, in the size regime of 2.0-3.3 nm, were prepared in the cubic phase by a wet chemical method. Five organic thiols were used for capping: (i) 1,4-dithiothreitol (DTT), (ii) 2-mercaptoethanol (ME), (iii) cysteine (Cys), (iv) methionine (Meth), and (v) glutathione (GSH). Structural studies were carried out by x-ray diffraction (XRD) and transmission electron microscopy (TEM), which revealed the cubic phase of CdS. Optical properties were studied by FT-IR, UV-visible and fluorescence spectroscopic techniques, and a comparison was made between uncapped and capped CdS. FT-IR studies suggested two different bonding mechanisms of the capping agents with the CdS. GSH and DTT capped CdS showed significant decrease in absorption wavelengths. An increase in band gap was observed in two cases: when (i) capped and (ii) decreased in size. The band gap was increased from 2.50 eV for the uncapped to 2.77 eV for the DTT capped CdS. DTT was found to be the best capping agent for CdS among these five organic thiols in two aspects: (i) yielding lower grain size in cubic phase, and (ii) good fluorescence properties with efficient quenching of the surface traps.

2008-10-29

364

Preparation of nanometer composite affinity membrane used for rapidly and efficiently separating and purifying thiol protease  

UK PubMed Central (United Kingdom)

The invention relates to a preparation of a nanometer composite affinity membrane used for rapidly and efficiently separating and purifying thiol protease, which comprises the steps of: (1) using a mixed solution of hexafluoroisopropanol (HFIP) and formic acid as the solvent system and doping the mixed solution in a reactor (2) adding polyamide 6 and chitosan powder in the reactor (3) putting the reactor in a water bath oscillator to obtain polyamide 6/chitosan spinning solution (4) using the spinning solution to perform the electrospinning to prepare a nanometer fiber membrane and then drying the nanometer fiber membrane (5) cutting the nanometer fiber membrane into circular membranes and then activating the circular membranes (6) putting the circular membranes into dye solution for reaction after a plurality of times of washing (7) adding NaCl solution for treatment (8) adding Na2CO3 for fixation reaction and (9) cooling and washing. The preparation of the nanometer composite affinity membrane used for rapidly and efficiently separating and purifying thiol protease has the advantages of simple operation, short time and large adsorbing capacity the prepared nanometer affinity membrane is rapid, simple, cheap and efficient, can efficiently purify the thiol protease and is applicable to scale production.

LIMIN ZHU; HAITAO ZHANG; JUNFANG LIU; MINJUN CAO; HUALI NIE

365

Enhancement of volatile thiol release of Saccharomyces cerevisiae strains using molecular breeding.  

Science.gov (United States)

Cysteine-conjugated volatile thiols are powerful aromatic compounds that contribute to the fruity notes of many white wines and especially Sauvignon Blanc. Genetic selection programs of wine yeast starters able to produce more volatile thiols constitute, therefore, an important goal for the wine industry. Recent investigations on yeast metabolism suggested that the ß-lyase Irc7p and the control of its gene expression by nitrogen catabolite repression constitute a rational way for yeast genetic improvement. This work demonstrates that the use of a natural ure2 mutation can be used to design wine starters with an enhanced capacity of volatile thiols production. By applying backcrosses driven by molecular markers, this allelic form was introduced in different starter backgrounds. Our investigations demonstrate that the ure2 inheritance is able to enhance the production of 4MMP (recently renamed 4MSP) and 3MH (recently renamed 3SH). For 4MMP, this effect depends of the presence of the allele IRC7LT encoding a long form of the Irc7 protein. Moreover, a correlation in between the expression level of this allelic form and 4MMP production was found within industrial starters. All together, these results emphasised the use of molecular breeding for improving quantitative traits of industrial strains without the use of genetically modifying strategies. PMID:23423325

Dufour, Matthieu; Zimmer, Adrien; Thibon, Cécile; Marullo, Philippe

2013-02-20

366

Enhancement of volatile thiol release of Saccharomyces cerevisiae strains using molecular breeding.  

UK PubMed Central (United Kingdom)

Cysteine-conjugated volatile thiols are powerful aromatic compounds that contribute to the fruity notes of many white wines and especially Sauvignon Blanc. Genetic selection programs of wine yeast starters able to produce more volatile thiols constitute, therefore, an important goal for the wine industry. Recent investigations on yeast metabolism suggested that the ß-lyase Irc7p and the control of its gene expression by nitrogen catabolite repression constitute a rational way for yeast genetic improvement. This work demonstrates that the use of a natural ure2 mutation can be used to design wine starters with an enhanced capacity of volatile thiols production. By applying backcrosses driven by molecular markers, this allelic form was introduced in different starter backgrounds. Our investigations demonstrate that the ure2 inheritance is able to enhance the production of 4MMP (recently renamed 4MSP) and 3MH (recently renamed 3SH). For 4MMP, this effect depends of the presence of the allele IRC7LT encoding a long form of the Irc7 protein. Moreover, a correlation in between the expression level of this allelic form and 4MMP production was found within industrial starters. All together, these results emphasised the use of molecular breeding for improving quantitative traits of industrial strains without the use of genetically modifying strategies.

Dufour M; Zimmer A; Thibon C; Marullo P

2013-07-01

367

Low-molecular-weight thiol-dependent antioxidant and antinitrosative defences in Salmonella pathogenesis.  

Science.gov (United States)

We found herein that the intracytoplasmic pool of the low-molecular-weight (LMW) thiol glutathione (GSH) is readily oxidized in Salmonella exposed to nitric oxide (NO). The hypersusceptibility of gshA and gshB mutants lacking ?-glutamylcysteine and glutathione synthetases to NO and S-nitrosoglutathione indicates that GSH antagonizes the bacteriostatic activity of reactive nitrogen species. Metabolites of the GSH biosynthetic pathway do not affect the enzymatic activity of classical NO targets such as quinol oxidases. In contrast, LMW thiols diminish the nitrosative stress experienced by enzymes, such as glutamine oxoglutarate amidotransferase, that contain redox active cysteines. LMW thiols also preserve the transcription of Salmonella pathogenicity island 2 gene targets from the inhibitory activity of nitrogen oxides. These findings are consistent with the idea that GSH scavenges reactive nitrogen species (RNS) other than NO. Compared with the adaptive response afforded by inducible systems such as the hmp-encoded flavohaemoprotein, gshA, encoding the first step of GSH biosynthesis, is constitutively expressed in Salmonella. An acute model of salmonellosis has revealed that the antioxidant and antinitrosative properties associated with the GSH biosynthetic pathway represent a first line of Salmonella resistance against reactive oxygen and nitrogen species engendered in the context of a functional NRAMP1(R) divalent metal transporter. PMID:23217033

Song, Miryoung; Husain, Maroof; Jones-Carson, Jessica; Liu, Lin; Henard, Calvin A; Vázquez-Torres, Andrés

2012-12-21

368

Thiol-disulfide interchange in the tocinoic acid/glutathione system during freezing and drying.  

UK PubMed Central (United Kingdom)

Thiol-disulfide interchange ("disulfide scrambling") is a common mechanism of covalent aggregation for protein drugs. Using tocinoic acid (cyclo-S-Cys-Tyr-Ile-Gln-Asn-Cys-(S); TA(ox)) and glutathione (?Glu-Cys-Gly; GSH), our previous work demonstrated that thiol/disulfide interchange is affected by lyophilization in a manner consistent with irreversible and regioselective loss of TA(ox) (Zhang et al., 2009, J Pharm Sci 98/9: 3312-3318). Here, we explore the contributions of stages of the lyophilization cycle to perturbations in thiol/disulfide interchange in the TA/GSH system. TA(ox) and GSH were co-lyophilized from phosphate buffer in the presence or absence of various excipients, then analyzed for TA(ox) and mixed disulfide products by reverse phase high performance liquid chromatography (rp-HPLC). Perturbations were found to occur primarily during freezing, before significant amounts of ice were removed by sublimation. Addition of a lyoprotectant (sucrose), a cryoprotectant (Tween-20) and flash-freezing influenced the product distribution only while ice was still present. Decreasing the redox potential by the addition of oxidized glutathione (GSSG) affected the product distribution differently in lyophilized samples and solution controls, but in neither case led to increased conservation of TA(ox).

Thing M; Zhang J; Laurence J; Topp EM

2010-12-01

369

Catalyst for synthesizing methane thiol from synthetic gas containing high-concentration hydrogen sulfide  

UK PubMed Central (United Kingdom)

The invention relates to a catalyst to use synthetical gas containing high concentration H2S as raw material to synthesize methane thiol by one-step method, composed of carrier, active component and active accelerant, where the carrier selects SiO2, TiO2 or heavy rare earth oxide the active component is Mo-O-K based compound, converted by the fore body K2MoO4 or (NH4)6Mo7O24 plus sylvine or MoO3 plus sylvine the active accelerant is mainly transition metal like Mn, Fe, Co, Ni, Ce, La, etc, or rare earth oxide it makes catalysis reaction at 295 deg.C and 0.2 Mpa in the volume ratio of the raw material gases CL/H2/H2S=1/2/(0.1-1) at an airspeed of (1-5) x ten to the power 3 h-1, showing high activity and selectivity, the methane thiol's time-space catching rate is up to 0.18-0.25g.h-1. ml-1cat, and the methane thiol's selectivity is 93.5%-98.8%.

YANG YIQUAN; WANG QI; LIN RENCUN

370

Immunoprotection in sheep against Haemonchus contortus using its thiol-purified excretory/secretory proteins  

Directory of Open Access Journals (Sweden)

Full Text Available Excretory/Secretory antigen was prepared by culturing live adult worms of Haemonchus contortus in RPMI 1640 medium at a concentration of 50 worms per mL in a culture-flask at 37 ?C for 24 hr and the culture supernatant was used as antigen. The E/S antigen was purified by thiol-sepharose affinity chromatography. On western blot analysis, it was demonstrated that thiol-purified antigen showed a single reactive band at 66 kDa. In immunization trial, sheep were administered intramuscularly with 500 ?g of thiol-purified excretory/secretory antigen along with montanide as adjuvant on day 0, 30 and 60. On ELISA, it was observed that the mean absorbance values were significantly (p ? 0.01) higher up to 20 weeks post immunization in Group-I (purified antigen) compared to Group- II (unimmunized control). Further, the mean EPG values was lower in Group I (200.00 ± 40.82 to 400.00 ± 91.29) than Group II (2200.00 ± 108.01 to 5100.00 ± 169.56) and the percentage reduction in mean fecal egg counts was 88.50%. Similarly, the mean abomasal worm counts was lower in Group I (808.33 ± 78.29) than Group II (3280.00 ± 147.19) and the percentage reduction in mean abomasal worm count was 75.40%.

Selvarayar Arunkumar

2012-01-01

371

Brain PP2A is modified by thiol-disulfide exchange and intermolecular disulfide formation.  

UK PubMed Central (United Kingdom)

The regulation of protein phosphatase 2A (PP2A) activity by thiol-disulfide exchange and resulting formation of an intermolecular disulfide was examined following exposure of a rat brain soluble fraction to a biotinylated derivative of the model disulfide HPDP (HPDP-biotin) which would be expected to label reactive protein thiols with a disulfide-linked biotin. The results show that a low concentration (500 microM) of HPDP-biotin produced substantial inhibition of PP2A activity and promoted the binding of the catalytic subunit of PP2A to an immobilized avidin-affinity column. Both the inhibition of PP2A activity and the binding of PP2A to the avidin column were reversed by treatment with the disulfide reducing agent dithiothreitol (DTT). Furthermore, the specific activity of PP2A was up to 7-fold higher in the DTT-eluted fractions from the avidin-affinity column than in the soluble fraction. These findings demonstrate directly that PP2A is susceptible to reversible inhibitory modification by thiol-disulfide exchange and provide mechanistic support for the emerging view that PP2A is an oxidant-sensitive protein phosphatase.

Foley TD; Kintner ME

2005-05-01

372

Brain PP2A is modified by thiol-disulfide exchange and intermolecular disulfide formation.  

Science.gov (United States)

The regulation of protein phosphatase 2A (PP2A) activity by thiol-disulfide exchange and resulting formation of an intermolecular disulfide was examined following exposure of a rat brain soluble fraction to a biotinylated derivative of the model disulfide HPDP (HPDP-biotin) which would be expected to label reactive protein thiols with a disulfide-linked biotin. The results show that a low concentration (500 microM) of HPDP-biotin produced substantial inhibition of PP2A activity and promoted the binding of the catalytic subunit of PP2A to an immobilized avidin-affinity column. Both the inhibition of PP2A activity and the binding of PP2A to the avidin column were reversed by treatment with the disulfide reducing agent dithiothreitol (DTT). Furthermore, the specific activity of PP2A was up to 7-fold higher in the DTT-eluted fractions from the avidin-affinity column than in the soluble fraction. These findings demonstrate directly that PP2A is susceptible to reversible inhibitory modification by thiol-disulfide exchange and provide mechanistic support for the emerging view that PP2A is an oxidant-sensitive protein phosphatase. PMID:15823574

Foley, Timothy D; Kintner, Marissa E

2005-05-20

373

Preventing thiol-yne addition improves the specificity of strain-promoted azide-alkyne cycloaddition.  

UK PubMed Central (United Kingdom)

The 1,3-dipolar cycloaddition of azides with ring-strained alkynes is one of the few bioorthogonal reactions suitable for specific biomolecule labeling in complex biological systems. Nevertheless, azide-independent labeling of proteins by strained alkynes can occur to a varying extent, thereby limiting the sensitivity of assays based on strain-promoted azide-alkyne cycloaddition (SPAAC). In this study, a subset of three cyclooctynes, dibenzocyclooctyne (DIBO), azadibenzocyclooctyne (DIBAC), and bicyclo[6.1.0]nonyne (BCN), was used to evaluate the azide-independent labeling of proteins in vitro. For all three cyclooctynes, we show that thiol-yne addition with reduced peptidylcysteines is responsible for most of the azide-independent polypeptide labeling. The identity of the reaction product was confirmed by LC-MS and NMR analysis. Moreover, we show that undesired thiol-yne reactions can be prevented by alkylating peptidylcysteine thiols with iodoacetamide (IAM). Since IAM is compatible with SPAAC, a more specific azide-dependent labeling is achieved by preincubating proteins containing reduced cysteines with IAM.

van Geel R; Pruijn GJ; van Delft FL; Boelens WC

2012-03-01

374

Characterization of plasma thiol redox potential in a common marmoset model of aging.  

UK PubMed Central (United Kingdom)

Due to its short lifespan, ease of use and age-related pathologies that mirror those observed in humans, the common marmoset (Callithrix jacchus) is poised to become a standard nonhuman primate model of aging. Blood and extracellular fluid possess two major thiol-dependent redox nodes involving cysteine (Cys), cystine (CySS), glutathione (GSH) and glutathione disulfide (GSSG). Alteration in these plasma redox nodes significantly affects cellular physiology, and oxidation of the plasma Cys/CySS redox potential (E hCySS) is associated with aging and disease risk in humans. The purpose of this study was to determine age-related changes in plasma redox metabolites and corresponding redox potentials (E h) to further validate the marmoset as a nonhuman primate model of aging. We measured plasma thiol redox states in marmosets and used existing human data with multivariate adaptive regression splines (MARS) to model the relationships between age and redox metabolites. A classification accuracy of 70.2% and an AUC of 0.703 were achieved using the MARS model built from the marmoset redox data to classify the human samples as young or old. These results show that common marmosets provide a useful model for thiol redox biology of aging.

Roede JR; Uppal K; Liang Y; Promislow DE; Wachtman LM; Jones DP

2013-01-01

375

Superhydrophobic hybrid inorganic-organic thiol-ene surfaces fabricated via spray-deposition and photopolymerization.  

UK PubMed Central (United Kingdom)

We report a simple and versatile method for the fabrication of superhydrophobic inorganic-organic thiol-ene coatings via sequential spray-deposition and photopolymerization under ambient conditions. The coatings are obtained by spray-deposition of UV-curable hybrid inorganic-organic thiol-ene resins consisting of pentaerythritol tetra(3-mercaptopropionate) (PETMP), triallyl isocyanurate (TTT), 2,4,6,8-tetramethyl-2,4,6,8-tetravinylcyclotetrasiloxane (TMTVSi), and hydrophobic fumed silica nanoparticles. The spray-deposition process and nanoparticle agglomeration/dispersion provide surfaces with hierarchical morphologies exhibiting both micro- and nanoscale roughness. The wetting behavior, dependent on the concentration of TMTVSi and hydrophobic silica nanoparticles, can be varied over a broad range to ultimately provide coatings with high static water contact angles (>150°), low contact angle hysteresis, and low roll off angles (<5°). The cross-linked thiol-ene coatings are solvent resistant, stable at low and high pH, and maintain superhydrophobic wetting behavior after extended exposure to elevated temperatures. We demonstrate the versatility of the spray-deposition and UV-cure process on a variety of substrate surfaces including glass, paper, stone, and cotton fabric.

Sparks BJ; Hoff EF; Xiong L; Goetz JT; Patton DL

2013-03-01

376

Thiol-reactive compounds from garlic inhibit the epithelial sodium channel (ENaC).  

Science.gov (United States)

The epithelial sodium channel (ENaC) is a key factor in the transepithelial movement of sodium, and consequently salt and water homeostasis in various organs. Dysregulated activity of ENaC is associated with human diseases such as hypertension, the salt-wasting syndrome pseudohypoaldosteronism type 1, cystic fibrosis, pulmonary oedema or intestinal disorders. Therefore it is important to identify novel compounds that affect ENaC activity. This study investigated if garlic (Allium sativum) and its characteristic organosulfur compounds have impact on ENaCs. Human ENaCs were heterologously expressed in Xenopus oocytes and their activity was measured as transmembrane currents by the two-electrode voltage-clamp technique. The application of freshly prepared extract from 5g of fresh garlic (1% final concentration) decreased transmembrane currents of ENaC-expressing oocytes within 10 min. This effect was dose-dependent and irreversible. It was fully sensitive to the ENaC-inhibitor amiloride and was not apparent on native control oocytes. The effect of garlic was blocked by dithiothreitol and l-cysteine indicating involvement of thiol-reactive compounds. The garlic organosulsur compounds S-allylcysteine, alliin and diallyl sulfides had no effect on ENaC. By contrast, the thiol-reactive garlic compound allicin significantly inhibited ENaC to a similar extent as garlic extract. These data indicate that thiol-reactive compounds which are present in garlic inhibit ENaC. PMID:22668601

Krumm, Patrick; Giraldez, Teresa; Alvarez de la Rosa, Diego; Clauss, Wolfgang G; Fronius, Martin; Althaus, Mike

2012-05-17

377

Thiol-reactive compounds from garlic inhibit the epithelial sodium channel (ENaC).  

UK PubMed Central (United Kingdom)

The epithelial sodium channel (ENaC) is a key factor in the transepithelial movement of sodium, and consequently salt and water homeostasis in various organs. Dysregulated activity of ENaC is associated with human diseases such as hypertension, the salt-wasting syndrome pseudohypoaldosteronism type 1, cystic fibrosis, pulmonary oedema or intestinal disorders. Therefore it is important to identify novel compounds that affect ENaC activity. This study investigated if garlic (Allium sativum) and its characteristic organosulfur compounds have impact on ENaCs. Human ENaCs were heterologously expressed in Xenopus oocytes and their activity was measured as transmembrane currents by the two-electrode voltage-clamp technique. The application of freshly prepared extract from 5g of fresh garlic (1% final concentration) decreased transmembrane currents of ENaC-expressing oocytes within 10 min. This effect was dose-dependent and irreversible. It was fully sensitive to the ENaC-inhibitor amiloride and was not apparent on native control oocytes. The effect of garlic was blocked by dithiothreitol and l-cysteine indicating involvement of thiol-reactive compounds. The garlic organosulsur compounds S-allylcysteine, alliin and diallyl sulfides had no effect on ENaC. By contrast, the thiol-reactive garlic compound allicin significantly inhibited ENaC to a similar extent as garlic extract. These data indicate that thiol-reactive compounds which are present in garlic inhibit ENaC.

Krumm P; Giraldez T; Alvarez de la Rosa D; Clauss WG; Fronius M; Althaus M

2012-07-01

378

Gold Nanoparticles Protected with Thiol-Derivatized Amphiphilic Poly(epsilon-caprolactone)-b-poly(acrylic acid)  

DEFF Research Database (Denmark)

Amphiphilic poly(epsilon-caprolactone)-b-poly(acrylic acid) (HS-PCL-b-PAA) with a thiol functionality in the PCL terminal has been prepared in a novel synthetic cascade. Initially, living anionic ring-opening polymerization (ROP) of epsilon-caprolactone (epsilon-CL) employing the difunctional initiator, 2-hydroxyethyl 2-bromoisobutyrate, followed by esterification with 2,4-dinitrophenyl- or 4-monomethoxytrityl-protected mercaptoacetic acids (Prot-), provided well-defined PCL macroinitiators capped with protected thiols. The macroinitiators allowed atom transfer radical polymerization (ATRP) of tent-butyl acrylate (tBA) in a controlled fashion by use of NiBr2(PPh3)(2) catalyst to produce Prot-PCL-b-PtBA with narrow polydispersities (1.17-1.39). Subsequent mild deprotection protocols provided HS-PCL-b-PAA. Reduction of a gold salt in the presence of this macroligand under thiol-deficient conditions afforded stable, aggregation-free nanoparticles, as evidenced from UV-vis spectroscopy and transmission electron microscopy (TEM), the latter revealed nanoparticles with a mean diameter of 9.0 +/- 3.1 nm.

Javakhishvili, Irakli; Hvilsted, SØren

2009-01-01

379

Rapid approach to biobased telechelics through two one-pot thiol-ene click reactions.  

Science.gov (United States)

The application of environmentally friendly thiol-ene chemistry to the preparation of biobased telechelics is presented in this work. This methodology is based on two one-pot photoinitiated thiol-ene click processes: step-growth polymerization using a 3,6-dioxa-1,8-octanedithiol and end-group postpolymerization modification with three functional thiols: 2-mercaptoethanol, 3-mercaptopropionic acid, and 3-mercaptopropyltrimethoxysilane. We applied this approach to a potentially 100% biomass-derived monomer, allyl ester of 10-undecenoic acid (UDA). To show the generality and scope of this methodology, a series of well-defined telechelics with molecular weight ranging from 1000-3000 g/mol and hydroxyl, carboxyl, or trimethoxysilyl groups at the polymer terminus were prepared. An exhaustive (1)H NMR and MALDI-TOF MS analyses demonstrates the highly end-group fidelity of this methodology being an interesting procedure for the accelerated preparation of telechelics derived from divinyl monomers. UDA-based thelechelic diol prepared using this methodology was reacted with 4,4'-methylenebis(phenylisocyanate) and 1,4-butanediol as the chain extender to obtain multiblock poly(ester urethane). PMID:20462176

Lluch, Cristina; Ronda, Joan C; Galià, Marina; Lligadas, Gerard; Cádiz, Virginia

2010-06-14

380

Rapid approach to biobased telechelics through two one-pot thiol-ene click reactions.  

UK PubMed Central (United Kingdom)

The application of environmentally friendly thiol-ene chemistry to the preparation of biobased telechelics is presented in this work. This methodology is based on two one-pot photoinitiated thiol-ene click processes: step-growth polymerization using a 3,6-dioxa-1,8-octanedithiol and end-group postpolymerization modification with three functional thiols: 2-mercaptoethanol, 3-mercaptopropionic acid, and 3-mercaptopropyltrimethoxysilane. We applied this approach to a potentially 100% biomass-derived monomer, allyl ester of 10-undecenoic acid (UDA). To show the generality and scope of this methodology, a series of well-defined telechelics with molecular weight ranging from 1000-3000 g/mol and hydroxyl, carboxyl, or trimethoxysilyl groups at the polymer terminus were prepared. An exhaustive (1)H NMR and MALDI-TOF MS analyses demonstrates the highly end-group fidelity of this methodology being an interesting procedure for the accelerated preparation of telechelics derived from divinyl monomers. UDA-based thelechelic diol prepared using this methodology was reacted with 4,4'-methylenebis(phenylisocyanate) and 1,4-butanediol as the chain extender to obtain multiblock poly(ester urethane).

Lluch C; Ronda JC; Galià M; Lligadas G; Cádiz V

2010-06-01

 
 
 
 
381

Optimization of Optical Properties of Polycarbonate Film with Thiol Gold-Nanoparticles  

Directory of Open Access Journals (Sweden)

Full Text Available A new nanostructured composite film based on thiol gold nanoparticles dispersed in polycarbonate and prepared by evaporating a solution of 1-dodecanthiol gold nanoparticles and polycarbonate was developed for applications as optical lenses. Lenses with superior mechanical properties, coloring and UV ray absorption and with the same transparency as the matrix were obtained. The supporting highly transparent polycarbonate matrix and the chloroform solution of thiol gold nanoparticles, 3 nm mean size, was mixed according to a doping protocol employing different concentrations of thiol gold nanoparticles vs. polycarbonate. The presence of nanoparticles in the polymer films was confirmed by the spectrophotometric detection of the characteristic absorbance marker peak at 540–580 nm. The nanostructured films obtained show a better coverage in the UV-vis range (250–450 nm) even at very low doping ratios, of the order of 1:1,000. These results offer a very promising approach towards the development of efficient nanostructured materials for applications to optical lenses.

Claudio Larosa; Enrico Stura; Roberto Eggenhöffner; Claudio Nicolini

2009-01-01

382

A Novel Strategy for Global Analysis of the Dynamic Thiol Redox Proteome*  

Science.gov (United States)

Nitroxidative stress in cells occurs mainly through the action of reactive nitrogen and oxygen species (RNOS) on protein thiol groups. Reactive nitrogen and oxygen species-mediated protein modifications are associated with pathophysiological states, but can also convey physiological signals. Identification of Cys residues that are modified by oxidative stimuli still poses technical challenges and these changes have never been statistically analyzed from a proteome-wide perspective. Here we show that GELSILOX, a method that combines a robust proteomics protocol with a new computational approach that analyzes variance at the peptide level, allows a simultaneous analysis of dynamic alterations in the redox state of Cys sites and of protein abundance. GELSILOX permits the characterization of the major endothelial redox targets of hydrogen peroxide in endothelial cells and reveals that hypoxia induces a significant increase in the status of oxidized thiols. GELSILOX also detected thiols that are redox-modified by ischemia-reperfusion in heart mitochondria and demonstrated that these alterations are abolished in ischemia-preconditioned animals.

Martinez-Acedo, Pablo; Nunez, Estefania; Gomez, Francisco J. Sanchez; Moreno, Margoth; Ramos, Elena; Izquierdo-Alvarez, Alicia; Miro-Casas, Elisabet; Mesa, Raquel; Rodriguez, Patricia; Martinez-Ruiz, Antonio; Dorado, David Garcia; Lamas, Santiago; Vazquez, Jesus

2012-01-01

383

Attachment of hydrogel microstructures and proteins to glass via thiol-terminated silanes.  

UK PubMed Central (United Kingdom)

Micropatterning strategies often call for attachment of non-fouling biomaterials and immobilization of proteins in order to create biosensing surfaces or to control cell-surface interactions. Our laboratory has made frequent use of hydrogel photolithography - a micropatterning process for immobilizing poly(ethylene glycol) (PEG) hydrogel microstructures on glass surfaces. In the present study we explored the use of thiolsilane as a coupling layer for both covalent anchoring of hydrogel microstructures and covalent immobilization of proteins on glass. These new surfaces were compared to acryl-silane functionalized glass slides that allowed covalent attachment of gels but only physical adsorption of proteins as well as surfaces containing a mixture of both functional groups. We observed comparable attachment and retention of hydrogel microstructures on acryl and thiol-terminated silanes. Ellipsometry studies revealed presence of significantly higher level of proteins on thiol-functionalized glass. Overall, our studies demonstrate that thiol-silane functionalized glass surfaces may be used to create complex micropatterned surfaces comprised of covalently attached hydrogels and proteins. This simple and effective surface modification strategy will be broadly applicable in cellular engineering and biosensing studies employing hydrogel micropatterns.

Seo JH; Shin DS; Mukundan P; Revzin A

2012-10-01

384

Attachment of hydrogel microstructures and proteins to glass via thiol-terminated silanes.  

Science.gov (United States)

Micropatterning strategies often call for attachment of non-fouling biomaterials and immobilization of proteins in order to create biosensing surfaces or to control cell-surface interactions. Our laboratory has made frequent use of hydrogel photolithography - a micropatterning process for immobilizing poly(ethylene glycol) (PEG) hydrogel microstructures on glass surfaces. In the present study we explored the use of thiolsilane as a coupling layer for both covalent anchoring of hydrogel microstructures and covalent immobilization of proteins on glass. These new surfaces were compared to acryl-silane functionalized glass slides that allowed covalent attachment of gels but only physical adsorption of proteins as well as surfaces containing a mixture of both functional groups. We observed comparable attachment and retention of hydrogel microstructures on acryl and thiol-terminated silanes. Ellipsometry studies revealed presence of significantly higher level of proteins on thiol-functionalized glass. Overall, our studies demonstrate that thiol-silane functionalized glass surfaces may be used to create complex micropatterned surfaces comprised of covalently attached hydrogels and proteins. This simple and effective surface modification strategy will be broadly applicable in cellular engineering and biosensing studies employing hydrogel micropatterns. PMID:22652352

Seo, Jeong Hyun; Shin, Dong-Sik; Mukundan, Priam; Revzin, Alexander

2012-04-27

385

Rose bengal dye on thiol-terminated bilayer for molecular devices.  

Science.gov (United States)

Recently, it has become increasingly important to control molecular layers, especially with regard to the formation of bilayers, in order to avoid electrical shorts in molecular electronics. In this paper, we report on the characterization of an in situ thiol-terminated bilayer that is formed by hydrogen bonding between the amine group of an aminoalkanethiol monolayer on a gold surface and the free amine group of aminoalkanethiolates in a bulk solution. We also report on the use of a rose bengal (RB) monolayer on a thiol-terminated bilayer for the purpose of application in a molecular memory device. Using surface-sensitive techniques such as grazing angle Fourier transform infrared (FT-IR) spectroscopy, quartz crystal microbalance (QCM) measurement, ellipsometry, X-ray photoelectron spectroscopy (XPS), and cyclic voltammetry (CV), we characterized a thiol-terminated bilayer (TUA-AUT) and an RB functionalized monolayer on a bilayered surface (RB-TUA-AUT). For a control experiment, we prepared a single RB monolayer attached by an ethanethiol group to a gold surface. In order to assess the feasibility of the present approach with respect to application in molecular electronics, we tested the switching property of the self-assembled monolayers (SAMs) using conducting-probe atomic force microscopy (CP-AFM). The RB monolayer on the bilayered surface exhibited hysteresis, while a single RB monolayer gave an electrical short. PMID:17373828

Bang, Gyeong Sook; Park, Jonghyurk; Lee, Junghyun; Choi, Nak-Jin; Baek, Hee Yoel; Lee, Hyoyoung

2007-03-21

386

Rose bengal dye on thiol-terminated bilayer for molecular devices.  

UK PubMed Central (United Kingdom)

Recently, it has become increasingly important to control molecular layers, especially with regard to the formation of bilayers, in order to avoid electrical shorts in molecular electronics. In this paper, we report on the characterization of an in situ thiol-terminated bilayer that is formed by hydrogen bonding between the amine group of an aminoalkanethiol monolayer on a gold surface and the free amine group of aminoalkanethiolates in a bulk solution. We also report on the use of a rose bengal (RB) monolayer on a thiol-terminated bilayer for the purpose of application in a molecular memory device. Using surface-sensitive techniques such as grazing angle Fourier transform infrared (FT-IR) spectroscopy, quartz crystal microbalance (QCM) measurement, ellipsometry, X-ray photoelectron spectroscopy (XPS), and cyclic voltammetry (CV), we characterized a thiol-terminated bilayer (TUA-AUT) and an RB functionalized monolayer on a bilayered surface (RB-TUA-AUT). For a control experiment, we prepared a single RB monolayer attached by an ethanethiol group to a gold surface. In order to assess the feasibility of the present approach with respect to application in molecular electronics, we tested the switching property of the self-assembled monolayers (SAMs) using conducting-probe atomic force microscopy (CP-AFM). The RB monolayer on the bilayered surface exhibited hysteresis, while a single RB monolayer gave an electrical short.

Bang GS; Park J; Lee J; Choi NJ; Baek HY; Lee H

2007-04-01

387

Comparison of photopolymerizable thiol-ene PEG and acrylate-based PEG hydrogels for cartilage development.  

Science.gov (United States)

When designing hydrogels for tissue regeneration, differences in polymerization mechanism and network structure have the potential to impact cellular behavior. Poly(ethylene glycol) hydrogels were formed by free-radical photopolymerization of acrylates (chain-growth) or thiol-norbornenes (step-growth) to investigate the impact of hydrogel system (polymerization mechanism and network structure) on the development of engineered tissue. Bovine chondrocytes were encapsulated in hydrogels and cultured under free swelling or dynamic compressive loading. In the acrylate system immediately after encapsulation chondrocytes exhibited high levels of intracellular ROS concomitant with a reduction in hydrogel compressive modulus and higher variability in cell deformation upon compressive strain; findings that were not observed in the thiol-norbornene system. Long-term the quantity of sulfated glycosaminoglycans and total collagen was greater in the acrylate system, but the quality resembled that of hypertrophic cartilage with positive staining for aggrecan, collagens I, II, and X and collagen catabolism. The thiol-norbornene system led to hyaline-like cartilage production especially under mechanical loading with positive staining for aggrecan and collagen II and minimal staining for collagens I and X and collagen catabolism. Findings from this study confirm that the polymerization mechanism and network structure have long-term effects on the quality of engineered cartilage, especially under mechanical loading. PMID:24060418

Roberts, Justine J; Bryant, Stephanie J

2013-09-20

388

Investigation into the Effect of Molds in Grasses on Their Content of Low Molecular Mass Thiols  

Directory of Open Access Journals (Sweden)

Full Text Available The aim of this study was to investigate the effect of molds on levels of low molecular mass thiols in grasses. For this purpose, the three grass species Lolium perenne, Festulolium pabulare and Festulolium braunii were cultivated and sampled during four months, from June to September. The same species were also grown under controlled conditions. High-performance liquid chromatography with electrochemical detection was used for quantification of cysteine, reduced (GSH) and oxidized (GSSG) glutathione, and phytochelatins (PC2, PC3, PC4 and PC5). Data were statistically processed and analyzed. Thiols were present in all examined grass species. The effect of fungicide treatments applied under field conditions on the content of the evaluated thiols was shown to be insignificant. Species influenced (p < 0.05) PC3 and GSSG content. F. pabulare, an intergeneric hybrid of drought- and fungi-resistant Festuca arundinacea, was comparable in PC3 content with L. perenne and F. braunii under field conditions. Under controlled conditions, however, F. pabulare had higher (p < 0.05) PC3 content than did L. perenne and F. braunii. Under field conditions, differences between the evaluated species were recorded only in GSSG content, but only sampling in June was significant. F. pabulare had higher (p < 0.05) GSSG content in June than did L. perenne and F. braunii.

Jiri Skladanka; Vojtech Adam; Ondrej Zitka; Olga Krystofova; Miroslava Beklova; Rene Kizek; Zdenek Havlicek; Petr Slama; Adam Nawrath

2012-01-01

389

Characterization of plasma thiol redox potential in a common marmoset model of aging.  

Science.gov (United States)

Due to its short lifespan, ease of use and age-related pathologies that mirror those observed in humans, the common marmoset (Callithrix jacchus) is poised to become a standard nonhuman primate model of aging. Blood and extracellular fluid possess two major thiol-dependent redox nodes involving cysteine (Cys), cystine (CySS), glutathione (GSH) and glutathione disulfide (GSSG). Alteration in these plasma redox nodes significantly affects cellular physiology, and oxidation of the plasma Cys/CySS redox potential (E hCySS) is associated with aging and disease risk in humans. The purpose of this study was to determine age-related changes in plasma redox metabolites and corresponding redox potentials (E h) to further validate the marmoset as a nonhuman primate model of aging. We measured plasma thiol redox states in marmosets and used existing human data with multivariate adaptive regression splines (MARS) to model the relationships between age and redox metabolites. A classification accuracy of 70.2% and an AUC of 0.703 were achieved using the MARS model built from the marmoset redox data to classify the human samples as young or old. These results show that common marmosets provide a useful model for thiol redox biology of aging. PMID:24024176

Roede, James R; Uppal, Karan; Liang, Yongliang; Promislow, Daniel E L; Wachtman, Lynn M; Jones, Dean P

2013-07-18

390

Characterization of plasma thiol redox potential in a common marmoset model of aging?  

Science.gov (United States)

Due to its short lifespan, ease of use and age-related pathologies that mirror those observed in humans, the common marmoset (Callithrix jacchus) is poised to become a standard nonhuman primate model of aging. Blood and extracellular fluid possess two major thiol-dependent redox nodes involving cysteine (Cys), cystine (CySS), glutathione (GSH) and glutathione disulfide (GSSG). Alteration in these plasma redox nodes significantly affects cellular physiology, and oxidation of the plasma Cys/CySS redox potential (EhCySS) is associated with aging and disease risk in humans. The purpose of this study was to determine age-related changes in plasma redox metabolites and corresponding redox potentials (Eh) to further validate the marmoset as a nonhuman primate model of aging. We measured plasma thiol redox states in marmosets and used existing human data with multivariate adaptive regression splines (MARS) to model the relationships between age and redox metabolites. A classification accuracy of 70.2% and an AUC of 0.703 were achieved using the MARS model built from the marmoset redox data to classify the human samples as young or old. These results show that common marmosets provide a useful model for thiol redox biology of aging.

Roede, James R.; Uppal, Karan; Liang, Yongliang; Promislow, Daniel E.L.; Wachtman, Lynn M.; Jones, Dean P.

2013-01-01

391

Density functional study of a typical thiol tethered on a gold surface: ruptures under normal or parallel stretch  

International Nuclear Information System (INIS)

[en] The mechanical and dynamical properties of a model Au(111)/thiol surface system were investigated by using localized atomic-type orbital density functional theory in the local density approximation. Relaxing the system gives a configuration where the sulfur atom forms covalent bonds to two adjacent gold atoms as the lowest energy structure. Investigations based on ab initio molecular dynamics simulations at 300, 350 and 370 K show that this tethering system is stable. The rupture behaviour between the thiol and the surface was studied by displacing the free end of the thiol. Calculated energy profiles show a process of multiple successive ruptures that account for experimental observations. The process features successive ruptures of the two Au-S bonds followed by the extraction of one S-bonded Au atom from the surface. The force required to rupture the thiol from the surface was found to be dependent on the direction in which the thiol was displaced, with values comparable with AFM measurements. These results aid the understanding of failure dynamics of Au(111)-thiol-tethered biosurfaces in microfluidic devices where fluidic shear and normal forces are of concern

2006-10-14

392

INHIBITION OF TYPE I 5?-REDUCTASE BY MEDICINAL PLANT EXTRACTS  

Directory of Open Access Journals (Sweden)

Full Text Available Type I 5?-reductase has been implicated in skin disorders such as acne, hirsutism and male pattern baldness and its inhibition offers a potential treatment for these disorders. The aim of this study was to investigate the inhibition of type I 5?-reductase activity by extracts from Indian medicinal plants. Plant extracts were screened and selected based on their ability to inhibit Propionibacterium acnes and Staphylococcus epidermidis. Since type I 5?-reductase metabolises testosterone to ?4-androstene-3, 17-dione, the activity of enzyme was determined using RIA for testosterone and ?4-androstene-3, 17-dione. It was found that methanolic extract of Embelia ribes was a potent inhibitor of type I 5?-reductase (IC50:100?g/mL). Extracts of Vitex negundo, Terminalia chebula, and Terminalia bellerica which also inhibited type I 5?-reductase (IC50: 200-390 ?g /mL). Therefore herbal formulation of these plant extracts may be used in the treatment of skin disorders involving type I 5?-reductase.

Patil Vijaya; Samuel Grace; Mirapurkar Shubhangi; R. Krishna Mohan; Dasgupta Debjani

2011-01-01