f1fo atp synthase: Topics by WorldWideScience.org

Double-lock ratchet mechanism revealing the role of ?SER-344 in F_oF₁ ATP synthase

ATP is synthesized by ATP synthase (F_OF₁-ATPase). Its rotary electromotor (F_O) translocates protons (in some organisms sodium cations) and generates torque to drive...Full Text Available

2011-03-08

Phosphorylation of sites 3 and 2 in rabbit skeletal muscle glycogen synthase by a multifunctional protein kinase (ATP-citrate lyase kinase)

In a majority of living organisms, F_oF₁ ATP synthase performs the fundamental process of ATP synthesis. Despite the simple net reaction formula, ADP + P_i → ATP + H₂O,...Full Text Available

2011-03-22

Activation and Stiffness of the Inhibited States of F₁-ATPase Probed by Single-molecule Manipulation

A multifunctional protein kinase, purified from rat liver as ATP-citrate lyase kinase, has been identified as a glycogen synthase kinase. This kinase catalyzed incorporation of up to 1.5 mol of and)2numberSPO4/mol of synthase subunit associated with a decrease in the glycogen synthase activity ratio from 0.85 to a value of 0.15. Approximately 65-70% of the TUPO4 was incorporated into site 3 and 30-35% into site 2 as determined by reverse phase high performance liquid chromatography. This multifunctional kinase was distinguished from glycogen synthase kinase-3 on the basis of nucleotide and protein substrate specificities. Since the phosphate contents in glycogen synthase of sites 3 and 2 are altered in diabetes and by insulin administration, the possible involvement of the multifunctional kinase was explored. Glycogen synthase purified from ...

1985-10-05

Regulation of starch synthesis in potato tubers

F₁-ATPase (F₁), a soluble portion of F_oF₁-ATP synthase (F_oF₁), is an ATP-driven motor in which γϵ subunits rotate in...Full Text Available

2010-04-09

Open top chamber experiment ``Edelmannshof``: carbohydrate and energy metabolism in needles and trunks from Norway spruce fumigated with unfiltered or charcoal-filtered air in comparison with untreated trees

Following tuber excision from the mother plant sucrose synthase activity fell from 3,120 to 960 nmol/g.f. wt./h within 7 days and starch synthesis ({sup 14}C sucrose incorporated into isolated discs) from 23 to 7 nmol/g.f. wt./h. While the maximum catalytic activity of sucrose synthase was more than sufficient to account for the observed rate of starch synthesis a maximum of 27% of sucrose incorporated by discs was converted into starch within 3 h. This compared with 80% conversion of {sup 14}C glucose incorporated. Tuber excision also reduced the rate of starch biosynthesis with glucose as a substrate (from 206 to 64 nmol/g.f. wt./h). The activities of UDPG-pyrophosphorylase, PPi-PFK, ATP-PFK, starch synthase and hexokinase (glucose or fructose substrates) were unaffected by tuber removal. ADPG pyrophosphorylase activity was reduced from 8,000 to 4,500 nmol/g.f. wt./h. Preliminary experiments indicate ...

1990-05-01

Vascular ATP-sensitive potassium channels are over-expressed and partially regulated by nitric oxide in experimental septic shock

As part of a five-year-lasting open-top-chamber experiment at the `Edelmannshof` site (Welzheimer Wald) we analyzed Norway spruce trees (Picea abies (L.) Karst.) with regard to carbohydrate and energy (ATP/ADP ratio) metabolism. End products, intermediates, and a regulator of carbon allocation (starch, sucrose, hexoses, fructose-2,6-bisphosphate) as well as enzyme activities involved in sucrose metabolism (sucrose-P synthase, sucrose synthase, acid invertase) were determined in needle and trunk samples of the different treatments (charcoal-filtered, unfiltered, ambient air). Changes of these parameters were documented annually and with regard to needle development (up to 2 years after bud break), and diurnally during an ozone period in midsummer. Samples from trees grown in an unfiltered atmosphere showed relatively high sucrose and hexose contents of the needles. The capacities of sucrose-P synthase ...

1997-12-01

British Library Electronic Table of Contents (United Kingdom)

Purpose To study the activation and expression of vascular (aorta and small mesenteric arteries) potassium channels during septic shock with or without modulation of the NO pathway. Methods Septic shock was induced in rats by peritonitis. Selective inhibitors of vascular KATP (PNU-37883A) or BKCa [iberiotoxin (IbTX)] channels were used to demonstrate their involvement in vascular hyporeactivity. Vascular response to phenylephrine was measured on aorta and small mesenteric arteries mounted on a wire myograph. Vascular expression of potassium channels was studied by PCR and Western blot, in the presence or absence of 1400W, an inducible NO synthase (iNOS) inhibitor. Aortic activation of the transcriptional factor nuclear factor-kappaB (NF-?B) was assessed by electrophoretic mobility shift as...

2011-01-01

Photolabelling kinetics of ATP-ase of sarcoplasmatic reticulum by 3'-ARILAZIDO-ATP

Supramolecular Structures for Photochemical Energy Conversion

... atp-ase azido compounds biochemical reaction kinetics endoplasmic reticulum

1984-05-02

Extracellular ATP4- promotes cation fluxes in the J774 mouse macrophage cell line

OAK B188 The goal of this project is to mimic the energy transduction processes by which photosynthetic organisms harvest sunlight and convert it to forms of energy that are more easily used and stored. The results may lead to new technologies for solar energy harvesting based on the natural photosynthetic process. They may also enrich our understanding and control of photosynthesis in living organisms, and lead to methods for increasing natural biomass production, carbon dioxide removal, and oxygen generation. In our work to date, we have learned how to make synthetic antenna and reaction center molecules that absorb light and undergo photoinduced electron transfer to generate long-lived, energetic charge-separated states. We have assembled a prototype system in which artificial reaction centers are inserted into liposomes (artificial cell-like constructs), where they carry out light-driven transmembrane translocation of hydrogen ions to generate proton motive force. By insertion of ...

2003-08-26

Molecular Cloning and Characterization of the Yew Gene Encoding Squalene Synthase from Taxus cuspidata

Extracellular ATP stimulates transmembrane ion fluxes in the mouse macrophage cell line J774. In the presence of Mg2+, nonhydrolyzable ATP analogs and other purine and pyrimidine nucleotides do not elicit this response, suggesting the presence of a specific receptor for ATP on the macrophage plasma membrane. One candidate for such a receptor is the ecto-ATPase expressed on these cells. We, therefore, investigated the role of this enzyme in ATP-induced /sup 86/Rb+ efflux in J774 cells. The ecto-ATPase had a broad nucleotide specificity and did not hydrolyze extracellular ATP in the absence of divalent cations. /sup 86/Rb+ efflux was not blocked by inhibition of the ecto-ATPase and did not require Ca2+ or Mg2+. In fact, ATP-stimulated /sup 86/Rb+ efflux was inhibited by Mg2+ and correlated with the availability of ATP4- in the medium. In the ...

1987-03-05

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2007-09-01

Immobilization on Chitosan of a Thermophilic Trehalose Synthase from Thermus thermophilus HJ6

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2010-03-01

Cell Cycle-dependent Expression of Chitin Synthase Genes in Aspergillus nidulans

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2001-03-01

Two and 8-azido photoaffinity probes. 2. Studies on the binding process of 2-5A synthetase by photosensitive ATP analogues

The ATP-Binding Cassette Transporter ABCA4: Structural and Functional Properties and Role in Retinal Disease

The photoaffinity probes [#gamma#-"3"2P]2-azidoATP (2-N_3ATP) and [#alpha#-"3"2P]8-azido-ATP (8-N_3ATP) were used to investigate the binding of ATP to highly purified 2-5A synthetase. 2-N_3APT and 8-N_3ATP are substrates for 2-5A synthetase. In this study the authors show that 2- and 8-N_3ATP are competitive inhibitors of the enzymatic conversion of ATP to 2-5A. Ultraviolet irradiation results in the photoinsertion of 2-N_3ATP and 8-N_3ATP into the enzyme. The covalent photoinsertion of [#alpha#-"3"2P]8-N_3ATP into the 2-5A synthetase is proportional to the inactivation of the enzyme as UV irradiation is increased. Photolabeling of 2-5A synthetase is saturated at 1.5 mM 2-N_3ATP and 2.0 mM 8-N_3ATP. Computer analysis ...

ATP overflow in skeletal muscle 1A arterioles

ATP-binding cassette transporters (ABC transporters) utilize the energy of ATP hydrolysis to translocate an unusually diverse set of substrates across cellular membranes. ABCA4, also known as...Full Text Available

2010-01-01

ATP-gated P2X₃ receptors constitute a positive autocrine signal for insulin release in the human pancreatic ? cell

The purpose of this study was to investigate the sources of ATP in the 1A arteriole, and to investigate age-related changes in ATP overflow. Arterioles (1A) from the red portion of the gastrocnemius...Full Text Available

2010-08-15

A New Metal-Binding Site for Yeast Phosphoglycerate Kinase as Determined by the Use of a Metal-ATP Analog

Extracellular ATP has been proposed as a paracrine signal in rodent islets, but it is unclear what role ATP plays in human islets. We now show the presence of an ATP signaling pathway that enhances...Full Text Available

2010-04-06

Structure and Reaction Mechanism of Basil Eugenol Synthase

Suicide substrate β, γ-bidentate Rh(III)ATP (RhATP) was used to map the metal ion-binding site in yeast phosphoglycerate kinase (PGK). Cleavage of the RhATP-inactivated enzyme with...Full Text Available

1997-02-01

Partial Purification of Digitonin-Solubilized ?-Glucan Synthase from Red Beet Root ¹

Phenylpropenes, a large group of plant volatile compounds that serve in multiple roles in defense and pollinator attraction, contain a propenyl side chain. Eugenol synthase (EGS) catalyzes the reductive...Full Text Available

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Partial Purification and Characterization of Indol-3-Ylacetylglucosemyo-Inositol Indol-3-Ylacetyltransferase (Indoleacetic Acid-Inositol Synthase) ¹

An enriched glucan synthase fraction was obtained from red beet root microsomes by sequential extraction with the detergents 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate and digitonin....Full Text Available

1987-04-01

Expression in E. coli and characterization of the catalytic domain of Botrytis cinerea chitin synthase

A procedure is described for the purification of the enzyme indol-3-ylacetylglucose:myo-inositol indol-3-ylacetyltransferase (IAA-myo-inositol synthase). This enzyme...Full Text Available

1990-12-01

Differentially regulated malate synthase genes participate in carbon and nitrogen metabolism of S. cerevisiae.

BackgroundChitin synthase 3a (CHS3a) from Botrytis cinerea (Bc) catalyses the multiple transfer of N-acetylglucosamine (GlcNAc) residues to the...Full Text Available

Differential expression of prostaglandin-H synthase isoenzymes in normal and activated keratinocytes in vivo and in vitro.

We have isolated a second gene (MLS1), which in addition to DAL7, encodes malate synthase from S. cerevisiae. Expression of the two genes is specific for their physiological roles in carbon and nitrogen...Full Text Available

1992-11-11

Dexamethasone and nitric oxide synthase gene expression in brain.

Normal mouse epidermis constitutively expresses prostaglandin-H synthase 1 (PGHS-1) but no PGHS-2. Acute inflammation and epidermal hyperplasia, (hyperplastic transformation), as evoked in adult mouse...Full Text Available

1995-07-01

Cottonseed Malate Synthase ¹

Systemic administration of lipopolysaccharide (LPS), which causes endotoxemia and systemic inflammation, has been reported to induce expression of the gene for type II inducible nitric oxide synthase...Full Text Available

1997-03-01

Carotenoid Crystal Formation in Arabidopsis and Carrot Roots Caused by Increased Phytoene Synthase Protein Levels

Malate synthase (EC 4.1.3.2), an enzyme unique to the glyoxylate cycle, was purified to homogeneity from cotyledons of 72-hours, darkgrown cotton (Gossypium hirsutum L.) seedlings....Full Text Available

1987-08-01

Carbon Partitioning in Eelgrass (Regulation by Photosynthesis and the Response to Daily Light-Dark Cycles).

BackgroundAs the first pathway-specific enzyme in carotenoid biosynthesis, phytoene synthase (PSY) is a prime regulatory target. This includes a number of biotechnological approaches...Full Text Available

Acetolactate Synthase Activity in Developing Maize (Zea mays L.) Kernels

Diel variations in rates of C export, sucrose-phosphate synthase (SPS) and sucrose synthase (SS) activity, and C reserves were investigated in Zostera marina L. (eelgrass) to elucidate the environmental...Full Text Available

1995-08-01

Terpenoid-based defenses in conifers: cDNA cloning, characterization, and functional expression of wound-inducible (E)-?-bisabolene synthase from grand fir (Abies grandis)

Acetolactate synthase (EC 4.1.3.18) activity was examined in maize (Zea mays L.) endosperm and embryos as a function of kernel development. When assayed using unpurified homogenates,...Full Text Available

1988-01-01

Contraction-mediated inactivation of glycogen synthase is accompanied by inactivation of glycogen synthase phosphatase in human skeletal muscle.

(E)-α-Bisabolene synthase is one of two wound-inducible sesquiterpene synthases of grand fir (Abies grandis), and the olefin product of this cyclization reaction...Full Text Available

1998-06-09

Mechanism of adenylate kinase. Dose adenosine 5'-triphosphate bind to the adenosine 5'-monophosphate site

Activities of glycogen synthase (GS) and GS phosphatase were determined on human muscle biopsies before and after isometric contraction at 2/3 maximal voluntary force. Total GS activity did not change...Full Text Available

1989-05-01

Matching ATP Supply and Demand in Mammalian Heart: In Vivo, In Vitro and In Silico Perspectives

Although the subtrate binding properties of adenylate kinase (AK) have been studied extensively by various biochemical and biophysical techniques, it remains controversial whether uncomplexed adenosine 5'-triphosphate (ATP) binds to the adenosine 5'-monophosphate (AMP) site of AK. The authors present two sets of experiments which argue against binding of ATP to the AMP site. (a) /sup 31/P nuclear magnetic resonance titration of ATP with AK indicated a 1:1 stoichiometry on the basis of changes in coupling constants and line widths. This ruled out binding of ATP to both sites. (b) ATP and MgATP were found to behave similarly by protecting AK from spontaneous inactivation while AMP showed only a small degree of protection. Such inactivation could also be protected or reversed by dithioerythritol and is most likely due to oxidation of sulfhydryl ...

1987-10-06

Determination of Adenosine Triphosphate on Marine Particulates:Synthesis of Methods for Use on OTEC Samples

Although the heart rapidly adapts cardiac output to match the body’s circulatory demands, the regulatory mechanisms ensuring that sufficient ATP is available to perform the required...Full Text Available

2010-02-01

Radiolabeling of a wound-inducible pyridoxal phosphate utilizing protein from tomato: evidence for its identification as ACC synthase

Adenosine triphosphate (ATP) is an indicator of living biomass in marine particulates. This report details the method used by Lawrence Berkeley Laboratory to analyze particulate ATP in samples taken from oligotrophic, tropical ocean waters. It represents a synthesis of previously published methods.

1982-08-01

Biosynthesis of hyaluronan: direction of chain elongation

Aminocyclopropane 1-carboxylic acid (ACC) synthase, a pyridoxal phosphate utilizing enzyme, catalyzes the conversion of S-adenosylmethionine to ACC, the rate limiting step in the biosynthesis of the plant hormone, ethylene. Ethylene, besides being involved in normal plant growth processes, is also produced in response to stress, e.g. wounding, pathogen infection, etc. The authors report the partial purification (400 fold) of ACC synthase from wounded pink tomato pericarp by classical techniques including ammonium sulfate precipitation, ion exchange and phenyl sepharose chromatography. Further purification results in a decrease in specific activity apparently due to the instability of the enzyme and the low levels present in plant tissue. Radiolabeling of a pyridoxal phosphate-utilizing protein in the ACC synthase enriched fraction was achieved. Evidence that this radiolabeled protein is ACC synthase ...

1986-06-08

A high-performance liquid chromatography-based radiometric assay for sucrose-phosphate synthase and other UDP-glucose requiring enzymes

The mechanism of hyaluronan biosynthesis in vertebrates had been proposed to occur at the reducing end of growing chains. This mechanism was questioned because a recombinant synthase appeared to add...Full Text Available

2006-09-15

Thio Wax Ester Biosynthesis Utilizing the Unspecific Bifunctional Wax Ester Synthase/Acyl Coenzyme A:Diacylglycerol Acyltransferase of Acinetobacter sp. Strain ADP1

A method for product analysis that eliminates a problematic step in the radiometric sucrose-phosphate synthase assay is described. The method uses chromatography on a boronate-derivatized high-performance liquid chromatography column to separate the labeled product, (14C)sucrose phosphate, from unreacted uridine 5{prime}-diphosphate-(14C)glucose (UDP-Glc). Direct separation of these compounds eliminates the need for treatment of the reaction mixtures with alkaline phosphatase, thereby avoiding the problem of high background caused by contaminating phosphodiesterase activity in alkaline phosphatase preparations. The method presented in this paper can be applied to many UDP-Glc requiring enzymes; here the authors show its use for determining the activities of sucrose-phosphate synthase, sucrose synthase, and uridine diphosphate-glucose pyrophosphorylase in plant extracts.

1991-05-01

Pharmacokinetics and pharmacodynamics of terbogrel, a combined thromboxane A₂ receptor and synthase inhibitor, in healthy subjects

The bifunctional wax ester synthase/acyl coenzyme A (acyl-CoA):diacylglycerol acyltransferase (WS/DGAT) from Acinetobacter sp. strain ADP1 (formerly Acinetobacter calcoaceticus...Full Text Available

2005-02-01

Campylobacter jejuni Fatty Acid Synthase II: Structural and functional analysis of ?-hydroxyacyl-ACP dehydratase (FabZ)

AimsTo characterize the pharmacokinetics of terbogrel, a new combined thromboxane A₂ (TxA₂) receptor and synthase inhibitor, in healthy human subjects after...Full Text Available

2004-07-01

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Biological effects of exogenous adenosine 5 prime -triphosphate on cultured mammalian cells: Evidence for a receptor mechanism and its regulation by desensitization

Fatty acid biosynthesis is crucial for all living cells. In contrast to higher organisms, bacteria use a type II fatty acid synthase (FAS II) composed of a series of individual proteins, making...Full Text Available

2009-03-06

Adenosine triphosphate loading thallium-201 myocardial scintigraphy. Optimal dose and diagnostic accuracy

Exogenous adenosine 5{prime}-triphosphate (ATP) mobilized intracellular calcium in human carcinoma A43l cells and in Swiss 3T3 and 3T6 mouse fibroblasts by increasing inositol trisphosphate similar to well down growth factors (platelet-derived growth factor (PDGF), epidermal growth factor (EGF), bradykinin (BK), serum). Calcium mobilization was examined by video imaging of fura-2 fluorescence is single cells, following the radioactive isotope {sup 45}Ca, and monitoring the decrease in fluorescence of cells loaded with chlortetracycline. Uridine 5{prime}-triphosphate, but not other nucleotides, mimicked ATP. Single-cell analysis revealed synchronous responses in 10 sec to ATP, BK or serum, while PDGF (3T3) and EGF (A431) produced slower signals with significant cell-to-cell variation. PDGF desensitized 3T3 cells to ATP and BK added 100 sec later but ATP or BK did not desensitized to ...

1989-01-01

C-106 tank process ventilation test

Adenosine triphosphate (ATP) is an alternative to dipyridamole or adenosine in thallium-201 myocardial scintigraphy. However, the optimal dose of ATP has not been determined. A Doppler guide wire study showed the coronary flow velocity at a dose of 0.15 mg/kg of ATP was equal or higher than that at 0.14 mg/kg of adenosine or 0.56 mg/kg of dipyridamole. ATP was given intravenously to 67 patients with coronary artery disease at 0.15 mg/kg/min for 6 min. Thallium-201 was injected at 3 min, followed by immediate and delayed (3 hrs) tomographic imaging. There was no serious side effect during examination, although chest pain (26%), dyspnea (17%), and flushing (33%) were common. The sensitivity and specificity to detect coronary artery disease were 98 and 100%, respectively. The sensitivity to detect left anterior descending artery, left circumflex artery, and right coronary artery lesions was 94, 59 and 77%, ...

1995-01-01

Some ABCA3 mutations elevate ER stress and initiate apoptosis of lung epithelial cells

Project W-320 Acceptance Test Report for tank 241-C-106, 296-C-006 Ventilation System Acceptance Test Procedure (ATP) HNF-SD-W320-012, C-106 Tank Process Ventilation Test, was an in depth test of the 296-C-006 ventilation system and ventilation support systems required to perform the sluicing of tank C-106. Systems involved included electrical, instrumentation, chiller and HVAC. Tests began at component level, moved to loop level, up to system level and finally to an integrated systems level test. One criteria was to perform the test with the least amount of risk from a radioactive contamination potential stand point. To accomplish this a temporary configuration was designed that would simulate operation of the systems, without being connected directly to the waste tank air space. This was done by blanking off ducting to the tank and connecting temporary ducting and an inlet air filter and housing to the recirculation system. This configuration would eventually ...

1998-07-20

Radiosensitization of human lung fibroblasts by chemicals that decrease ATP levels

BackgroundABCA3 transporter (ATP-binding cassette transporter of the A subfamily) is localized to the limiting...Full Text Available

2011-01-01

P2Y receptors on astrocytes and microglia mediate opposite effects in astroglial proliferation

Radiosensitization by lactate, pyruvate, nalidixic acid and novobiocin was studied in exponentially growing SH-18L human lung fibroblasts. All the chemicals had a slight radiosensitizing effect at a low concentration and a definite effect at a higher one. Decreases in the D_0 and/or Dq values were present in each dose survival curve. Fibroblasts incubated with the low concentration of each chemical for 24 hrs after X irradiation showed no reduction in intracellular ATP content, whereas, the higher concentration produced a significant decrease. These observations suggest that the decrease in the ATP content may be involved in the radiosensitization of human fibroblasts at high concentrations of these chemicals. In contrast, radiosensitization at a low concentration is not explained by a relationship to ATP content. Different mechanisms may be involved in radiosensitization at low and high concentrations of these chemicals. ...

British Library Electronic Table of Contents (United Kingdom)

Nucleotides released upon brain injury signal to astrocytes and microglia playing an important role in astrogliosis, but the participation of microglia in the purinergic modulation of astrogliosis is still unclear. Highly enriched astroglial cultures and co-cultures of astrocytes and microglia were used to investigate the influence of microglia in the modulation of astroglial proliferation mediated by nucleotides. In highly enriched astroglial cultures, adenosine-5?-triphosphate (ATP), adenosine 5?-O-(3-thio)-triphosphate (ATP?S), adenosine 5?-O-(3-thio)-diphosphate (ADP?S; 0.01?1?mM), and adenosine-5?-diphosphate (ADP; 0.1?1?mM) increased proliferation up to 382%, an effect abolished in co-cultures containing 8% of microglia. The loss of ATP proliferative effect in co-cultures is supporte...

2011-01-01

Mitochondrial Potassium ATP Channels and Retinal Ischemic Preconditioning

Sucrose accumulation in mature sweet melon fruits. [Cucumis melo

PURPOSETo examine the mechanisms of ischemic preconditioning (IPC) related to the opening of mitochondrial KATP (mKATP) channels in the retina.Full Text Available

2006-05-01

Pore properties and pharmacological features of the P2X receptor channel in airway ciliated cells

Mesocarp tissue from sucrose-accumulating sweet melon (Cucumis melo cv. Galia) showed sucrose synthase activity (ca 1 nkat/gfw) while soluble acid invertase and sucrose phosphate synthase activities were not observed. Sucrose uptake into mesocarp discs was linear with sucrose concentration (1-500 mM) and unaffected by PCMBS and CCCP. Sucrose compartmentation into the vacuole also increased linearly with sucrose concentration as indicated by compartmental efflux kinetics. Mesocarp discs incubated in /sup 14/C-fructose + UDP-glu synthesized /sup 14/C-sucrose and efflux kinetics indicated that the /sup 14/C-sucrose was compartmentalized. These data support the hypothesis that two mechanisms are involved in sucrose accumulation in sweet melon: (1) compartmentation of intact sucrose and (2) synthesis of sucrose via sucrose synthase and subsequent compartmentation in the vacuole.

1987-04-01

Interaction of Extracellular Domain 2 of the Human Retina-specific ATP-binding Cassette Transporter (ABCA4) with All-trans-retinal*

Airway ciliated cells express an ATP-gated P2X receptor channel of unknown subunit composition (P2X_cilia) which is modulated by Na⁺ and by long exposures to ATP. P2X_cilia...Full Text Available

2006-03-15

Evaluation of indexes for tumor growth and effect of radiation therapy using MRS; Experimental studies on SCCVII tumor bearing mice

The retina-specific ATP-binding cassette (ABC) transporter, ABCA4, is essential for transport of all-trans-retinal from the rod outer segment discs in the retina and is associated with...Full Text Available

2010-06-18

Differential use of two cyclic electron flows around photosystem I for driving CO₂-concentration mechanism in C₄ photosynthesis

Changes in the phosphate metabolism of the transplanted SCCVII (squamous cell carcinoma VII) tumor were studied before and after irradiation using in vivo [sup 31]P-magnetic resonance spectroscopy (MRS). Phosphocreatine (PCr), [gamma]-, [alpha]-, [beta]-ATP, inorganic phosphate (Pi), phosphomonoester (PME), and phosphodiester (PDE) were detected in [sup 31]P-MR spectra of tumor tissues. The curve fitting method was employed for calculating each peak of [sup 31]P-MR spectra (PCr, [beta]-ATP, Pi, PME, PDE). The values of PCr, PCr/Pi, [beta]-ATP/Pi and pH decreased in proportion to the growth of tumor, whereas that of Pi, PME and PDE increased. But the changes of [beta]-ATP were minimal in degree. Based on the fact that the ratio of PCr/Pi highly correlated with tumor volume and showed the most remarkable change among various parameters, it might be the useful index for the tumor growth. Because the ...

1992-03-01

Diagnostic value of adenosine-triphosphate radionuclide ventriculography in coronary artery disease. Comparison with conventional exercise stress method

Whereas linear electron flow (LEF) in photosynthesis produces both ATP and NADPH, the cyclic electron flow (CEF) around photosystem I has been shown to produce only ATP. Two alternative routes have...Full Text Available

2005-11-15

ATP Hydrolysis in Eg5 Kinesin Involves a Catalytic Two-water Mechanism*^?

The purpose of this study was to assess the effect of adenosine-triphosphate (ATP) on hemodynamics and left ventricular (LV) function, and to examine the value of ATP radionuclide-ventriculography (RNVG) for detecting coronary artery disease (CAD) in comparison with standard exercise RNVG. Thirty-eight consecutive patients with suspected CAD underwent ATP (0.18 mg/Kg/min) RNVG and symptom-limited exercise RNVG. Coronary angiography revealed 27 patients with CAD (Group S) and 11 without CAD (Group N). In group S, exercise induced an increase in LV end-diastolic volume (LVEDV) and LV end-systolic volume (LVESV), and a decrease in LV ejection fraction (LVEF). In contrast, ATP infusion caused LVESV to decrease in both groups, improving LVEF. Only 14 of 39 segments whose wall motion was deteriorated by exercise showed worsening regional wall motion during ATP infusion. The sensitivity ...

1995-07-01

Transient increase in glucose 1,6-bisphosphate in human skeletal muscle during isometric contraction.

Motor proteins couple steps in ATP binding and hydrolysis to conformational switching both in and remote from the active site. In our kinesin·AMPPPNP crystal structure, closure of the active...Full Text Available

2010-02-19

The endogenous hydrogen sulfide producing enzyme cystathionine-? synthase contributes to visceral hypersensitivity in a rat model of irritable bowel syndrome

Changes in glucose 1,6-bisphosphate and regulators of glucose-1,6-bisphosphate synthase and phosphatase during isometric contraction have been determined. Biopsies were obtained from the quadriceps...Full Text Available

1989-03-15

Prostaglandin H₂ synthase-1 and -2 expression in guinea pig gestational tissues during late pregnancy and parturition

BackgroundThe pathogenesis of visceral hypersensitivity, a characteristic pathophysiological feature of irritable bowel syndrome (IBS), remains elusive. Recent studies suggest a...Full Text Available

Polyketide synthase chemistry does not direct biosynthetic divergence between 9- and 10-membered enediynes

Increased intrauterine prostaglandin (PG) production is crucial for the initiation of parturition. To investigate the mechanisms controlling intrauterine PG synthesis, we examined the expression of...Full Text Available

2005-12-15

Mycophenolic acid inhibits activation of inducible nitric oxide synthase in rodent fibroblasts

Enediynes are potent antitumor antibiotics that are classified as 9- or 10-membered according to the size of the enediyne core structure. However, almost nothing is known about enediyne core biosynthesis,...Full Text Available

2010-06-22

Molecular cloning and functional expression of geranylgeranyl pyrophosphate synthase from Coleus forskohlii Briq

Mycophenolate mofetil (MMF) is an immunosuppressive drug that acts as a selective inhibitor of inosine monophosphate dehydrogenase (IMPDH). MMF has recently been shown to inhibit the enzymatic activity...Full Text Available

2003-05-01

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Identification of Phloem Involved in Assimilate Loading in Leaves by the Activity of the Galactinol Synthase Promoter1

BackgroundIsopentenyl diphosphate (IPP), a common biosynthetic precursor to the labdane diterpene forskolin, has been biosynthesised via a non-mevalonate pathway. Geranylgeranyl...Full Text Available

Identification and Characterization of CPS1 as a Hyaluronic Acid Synthase Contributing to the Pathogenesis of Cryptococcus neoformans Infection?

The definition of “minor” veins in leaves is arbitrary and of uncertain biological significance. Generally, the term refers to the smallest vein classes in the leaf, believed to function...Full Text Available

2000-07-01

Genetic Association Analysis of NOS1 and Methamphetamine-Induced Psychosis Among Japanese

Cryptococcus neoformans is a pathogenic yeast that often causes devastating meningoencephalitis in immunocompromised individuals. We have previously identified the C. neoformans...Full Text Available

2007-08-01

Endogenous Nitric Oxide Protects Bacteria Against a Wide Spectrum of Antibiotics

The neuronal nitric oxide synthase gene (NOS1) is located at 12q24, a susceptibility region for schizophrenia, and produces nitric oxide (NO). NO has been reported to play important...Full Text Available

2011-03-01

Detection and Identification of Bartonella Species Pathogenic for Humans by PCR Amplification Targeting the Riboflavin Synthase Gene (ribC)

Bacterial nitric oxide synthases (bNOS) are present in many Gram-positive species and have been demonstrated to synthesize NO from arginine in vitro and in vivo. However, the physiological role...Full Text Available

2009-09-11

Characterization of Geraniol Synthase from the Peltate Glands of Sweet Basil1

Several Bartonella species have now been implicated as human pathogens. The recovery of these fastidious organisms in the clinical microbiology laboratory remains difficult, and current...Full Text Available

2003-03-01

A Bifunctional Geranyl and Geranylgeranyl Diphosphate Synthase Is Involved in Terpene Oleoresin Formation in Picea abies1[W][OA

The monoterpene fraction of the lemon-scented sweet basil (Ocimum basilicum) cv Sweet Dani consists mostly of citral (a mixture of geranial and neral), with lower levels of geraniol...Full Text Available

2004-01-01

Does adenine incorporation into nucleic acids measure total microbial production:: a response to comments by Fuhrman et al

The conifer Picea abies (Norway spruce) defends itself against herbivores and pathogens with a terpenoid-based oleoresin composed chiefly of monoterpenes (C₁₀) and diterpenes...Full Text Available

2010-02-01

Depression of calcium pump activity in renal cortex of vitamin D-deficient rats with secondary hyperparathyroidism

The uptake and incorporation of (/sup 3/H)adenine as a measure of total microbial (bacteria and unicellular algae) nucleic acid synthesis depends on the validity of several assumptions. A unique characteristic of the (/sup 3/H)adenine method is the ability to measure the specific radioactivity (nCi pmol/sup -1/) of the immediate precursor pool of the adenine incorporated into nucleic acids. This measurement permits correction for isotope dilution when the added radiotracer mixes with exogenous pools of structurally related compounds before uptake and with endogenous pools after transport into the cells. The immediate precursors to (/sup 3/H)adenine incorporation into cellular RNA and DNA are ATP and dATP. Since the intracellular dATP pool is in isotopic equilibrium with the ATP pool, one can determine the specific radioactivity of both precursor pools by measuring that of the intracellular ...

1986-11-01

Depression of calcium pump activity in renal cortex of vitamin D-deficient rats with secondary hyperparathyroidism

To examine the hormonal regulation of the ATP-dependent Ca{sup 2+} pump in the kidneys, the ATP-dependent Ca{sup 2+} uptake by the basolateral membrane vesicles in the renal cortex was measured using radioactive calcium ({sup 45}Ca{sup 2+}) in rats with vitamin D deficiency or rats undergoing thyroparathyroidectomy. The V{sub max} of the Ca{sup 2+} pump activity was increased not only by administering calcitriol, but also by normalizing the serum calcium level in vitamin D-deficient rats. PTH suppressed the Ca{sup 2+} pump activity in normocalcemic vitamin D-deficient rats. Thyroparathyroidectomy did not affect the Ca{sup 2+} pump activity in the kidneys of normal rats. It was concluded that the ATP-dependent Ca{sup 2+} pump activity was depressed by secondary hyperparathyroidism in vitamin D-deficient rats. (author).

1990-01-01

Triosephosphates Modulate Leaf Mitochondrial Phosphorylation by Inhibition and Uncoupling of Electron Transport ¹

To examine the hormonal regulation of the ATP-dependent Ca"2"+ pump in the kidneys, the ATP-dependent Ca"2"+ uptake by the basolateral membrane vesicles in the renal cortex was measured using radioactive calcium ("4"5Ca"2"+) in rats with vitamin D deficiency or rats undergoing thyroparathyroidectomy. The V_m_a_x of the Ca"2"+ pump activity was increased not only by administering calcitriol, but also by normalizing the serum calcium level in vitamin D-deficient rats. PTH suppressed the Ca"2"+ pump activity in normocalcemic vitamin D-deficient rats. Thyroparathyroidectomy did not affect the Ca"2"+ pump activity in the kidneys of normal rats. It was concluded that the ATP-dependent Ca"2"+ pump activity was depressed by secondary hyperparathyroidism in vitamin D-deficient rats. (author).

Transport of Arginine and Aspartic Acid into Isolated Barley Mesophyll Vacuoles ¹

The effect of TP (triosephosphates:glyceraldehyde-3 phosphate, GAP, +dihydroxyacetone phosphate, DHAP) on respiration, phosphorylation and matrix ATP/ADP ratios of isolated oat mesophyll mitochondria...Full Text Available

1985-11-01

The efflux of a fluorescent probe is catalyzed by an ATP-driven extrusion system in Lactococcus lactis.

The transport of arginine into isolated barley (Hordeum vulgare L.) mesophyll vacuoles was investigated. In the absence of ATP, arginine uptake was saturable with a K_m...Full Text Available

1991-10-01

The accuracy and reverse effects of thallium myocardial SPECT using adenosine triphosphate loading in the diagnosis of coronary artery disease. Comparison with other loading methods semiquantitatively

Many bacteria, both gram positive and gram negative, extrude in an energy-dependent manner the fluorescent pH indicator 2',7'-bis-(2-carboxyethyl)-5[and -6]-carboxyfluorescein (BCECF) (D. Molenaar,...Full Text Available

1992-05-01

The DEXD/H-box RNA Helicase DDX19 Is Regulated by an ?-Helical Switch*S?

The adverse effects and diagnostic accuracy of thallium-201 myocardial SPECT after intravenous infusion of adenosine triphosphate (ATP) were studied and compared with SPECT examinations with other type of loading. The subjects of the study were 147 patients with or without coronary artery disease, who underwent some type of loading SPECT and coronary arteriography (CAG) within 30 days. Myocardial ischemia was evaluated qualitatively in SPECT and was compared with CAG for the diagnostic accuracy of coronary artery disease. The degree of myocardial uptake defect was also calculated semiquantitatively using visual scoring method and compared with coronary artery severity score. The adverse effects occurred in 46.7% of ATP loading SPECT which was more frequent that DIP loading SPECT, but all of them were transient and mild. As for the diagnostic ability the ATP loading SPECT was as effective as other type of loading in ...

1999-09-01

Rapid Phytochrome-mediated Changes in Adenosine 5?-Triphosphate Content of Etiolated Bean Buds ¹

DEXD/H-box RNA helicases couple ATP hydrolysis to RNA remodeling by an unknown mechanism. We used x-ray crystallography and biochemical analysis of the human DEXD/H-box...Full Text Available

2009-04-17

P2X purinoceptors mediate an endothelium-dependent hyperpolarization in longitudinal smooth muscle of anterior mesenteric artery in young chickens

This study was designed to determine the effects of red and far red irradiation on ATP metabolism in etiolated bean buds (Phaseolus vulgaris L. var. Red Kidney). Compared to dark controls,...Full Text Available

1974-01-01

Molecular characterization of cytoplasmic male sterility conditioned by Gossypium harknessii cytoplasm (CMS-D2) in upland cotton

Background and purpose:The chicken anterior mesenteric artery contains an outer longitudinal smooth muscle layer, whose neural regulation remains to be elucidated. ATP evokes a depolarization...Full Text Available

2009-10-01

British Library Electronic Table of Contents (United Kingdom)

Cytoplasmic male sterility (CMS) is a maternally inherited trait that fails to produce functional pollen grains. The CMS system is widely employed to facilitate the utilization of heterosis in major crops. However, little is known about the CMS associated genes in Upland cotton (Gossypium hirsutum). The objective of this study was to compare CMS cotton (CMS-D2) with the cytoplasm from G. harknessii and its isogenic maintainer line with the normal fertile Upland cotton cytoplasm to identify CMS-D2 specific gene(s) and to develop CMS-specific sequence characterized amplified region (SCAR) markers. Based on Southern blot analysis using 10 mitochondrial gene-specific probes (cob, cox2, atp6, atp9, nad3, cox3, atpA, cox1, nad6 and nad9), three probes (cox3, atpA, and nad6) revealed restriction ...

2011-01-01

Kinetics of nucleotide transport in rat heart mitochondria studied by a rapid filtration technique

Increased expression of cholesterol transporter ABCA1 is highly correlated with severity of dementia in AD hippocampus

A rapid filtration technique has been used to measure at room temperature the kinetics of ADP and ATP transport in rat heart mitochondria in the millisecond time range. Transport was stopped by cessation of the nucleotide supply, without the use of a transport inhibitor, thus avoiding any quenching delay. The kinetics of ({sup 14}C)ADP transport in energized mitochondria were apparently monophasic. The rate of transport of ({sup 14}C)ATP in energized mitochondria was 5-10 times lower than that of ({sup 14}C)ADP. Upon uncoupling, the rate of ({sup 14}C)ATP uptake was enhanced, and that of ({sup 14}C)ADP uptake was decreased. However, the two rates did not equalize, indicating that transport was not exclusively electrogenic. Transport of ({sup 14}C)ADP and ({sup 14}C)ATP by resting mitochondria followed biphasic kinetics. Depletion of nucleotides in resting mitochondria resulted in a greater decrease in ...

1990-10-01

1 2 3 4 5 6 7 8 9 10 11

Identification of genetic variation and haplotype structure of the canine ABCA4 gene for retinal disease association studies

To gain insight into ATP-binding cassette transporter A1 (ABCA1) function and its potential role in AD pathology, we analyzed the expression of the cholesterol transporter ABCA1...Full Text Available

2010-03-08

Gyrase B Inhibitor Impairs HIV-1 Replication by Targeting Hsp90 and the Capsid Protein*

Over 200 mutations in the retina specific member of the ATP-binding cassette transporter super-family (ABCA4) have been associated with a diverse group of human retinal diseases....Full Text Available

2010-10-01

Energy metabolism of the untrained muscle of elite runners as observed by 31P magnetic resonance spectroscopy: evidence suggesting a genetic endowment for endurance exercise.

Chemical genetics is an emerging approach to investigate the biology of host-pathogen interactions. We screened several inhibitors of ATP-dependent DNA motors and detected the gyrase B inhibitor coumermycin...Full Text Available

2010-12-10

Efficient Phagocytosis Requires Triacylglycerol Hydrolysis by Adipose Triglyceride Lipase*

The purpose of this study was to investigate whether genetically determined properties of muscle metabolism contribute to the exceptional physical endurance of world-class distance runners. ATP, phosphocreatine,...Full Text Available

1988-12-01

Diterpene Biosynthesis in Maize Seedlings in Response to Fungal Infection ¹

Macrophage phagocytosis is an essential biological process in host defense and requires large amounts of energy. To date, glucose is believed to represent the prime substrate for ATP production in macrophages....Full Text Available

2010-06-25

Cytosolic Phosphofructokinase from Spinach Leaves ¹

A cell-free system which catalyzes the biosynthesis of terpene hydrocarbons when supplemented with mevalonate, Mn²⁺, and ATP was prepared from the scutellum-embryonic axis region of maize...Full Text Available

1979-09-01

Conditional deletion of Abca3 in alveolar type II cells alters surfactant homeostasis in newborn and adult mice

Cytosolic ATP-phosphofructokinase (PFK) from spinach leaves (Spinacia oleracea L.) was inhibited by submillimolar concentrations of free Mg²⁺. The free Mg²⁺ concentration...Full Text Available

1989-08-01

Comparison of McAuley/fasting insulin indices with ATP III clinical criteria for the diagnosis of insulin resistance in type 2 diabetes mellitus

ATP-binding cassette A3 (ABCA3) is a lipid transport protein required for synthesis and storage of pulmonary surfactant in type II cells in the alveoli. Abca3 was conditionally deleted...Full Text Available

2010-05-01

Chemiosmotic model for plant mitochondria

Objective:To estimate the prevalence of insulin resistant syndrome (IRS) among newly diagnosed patients with type 2 diabetes and to test their validity against two indices...Full Text Available

2011-07-01

Anaerobic Growth Yields of Aerobacter cloacae and Escherichia coli

A balanced application of chemiosmosis to plant mitochondria is presented. Electron transport carriers are sequenced to achieve proton translocation across the inner membrane. The processes of proton translocation, use, and release are integrated to yield a description of the effective proton gradient available for metabolite transport and ATP synthesis. 27 references, 6 figures.

1983-12-01

Adenosine 5?-triphosphate sulphurylase from Saccharomyces cerevisiae

Aerobacter cloacae UW-C83 and Escherichia coli K-12 were grown under various anaerobic environments. Y_atp values were calculated by determination...Full Text Available

1967-10-01

A TRAP Transporter for Pyruvate and Other Monocarboxylate 2-Oxoacids in the Cyanobacterium Anabaena sp. Strain PCC 7120?

1. ATP sulphurylase from Saccharomyces cerevisiae was purified 140-fold by using heat treatment, DEAE-cellulose chromatography and Sepharose 6B gel filtration. 2. The enzyme was stable...Full Text Available

1973-07-01

Synthesis of high specific activity tritium labelled [2-{sup 3}H]-adenosine-5`-triphosphate

In the cyanobacterium Anabaena sp. strain PCC 7120, open reading frames (ORFs) alr3026, alr3027, and all3028 encode a tripartite ATP-independent...Full Text Available

2010-11-01

Mapping of the nucleotide-binding sites in the ADP/ATP carrier of beef heart mitochondria by photolabeling with 2-azido[#alpha#-"3"2P]adenosine diphosphate

A procedure for high level tritium labelling at the C2-H position of adenosine 5`-triphosphate ([2-{sup 3}H]-ATP, 1), based on the tritiodehalogenation reaction of 2-bromoadenosine 5`-triphosphate (2) has been elaborated. This precursor was prepared in a six-step synthesis from guanosine. The tritiodehalogenation of (2) for three hours over palladium oxide in phosphate buffer yielded tritium labelled ATP with high specific activity, in good chemical yield. (author).

1996-08-01

Bacterial radiosensitization by using radiation processing in combination with essential oil: Mechanism of action

2-Azido[#alpha#-"3"2P]adenosine diphosphate (2-azido[#alpha#-"3"2P]ADP) has been used to photolabel the ADP/ATP carrier in beef heart mitochondria. In reversible binding assays carried out in the dark, this photoprobe was found to inhibit ADP/ATP transport in beef heart mitochondria and to bind to two types of specific sites of the ADP/ATP carrier characterized by high-affinity binding (K/sub d/ = 20 #mu#M) and low-affinity binding (K/sub d/ = 400 #mu#M). In contrast, it was unable to bind to specific carrier sites in inverted submitochondrial particles. Upon photoirradiation of beef heart mitochondria in the presence of 2-azido[#alpha#-"3"2P]ADP, the ADP/ATP carrier was covalently labeled. After purification, the photolabeled carrier protein was cleaved chemically by acidolysis or cyanogen bromide and enzymatically with the Staphylococcus aureus V8 protease. In the ADP/ATP carrier ...

Homology analyses of the protein sequences of fatty acid synthases from chicken liver, rat mammary gland, and yeast

Spice extracts under the form of essential oils were tested for their efficiency to increase the relative radiosensitivity of Listeria monocytogenes and Escherichia coli O157H7 in culture media. The two pathogens were treated by gamma-irradiation alone or in combination with oregano essential oil to evaluate their mechanism of action. The membrane murein composition, and the intracellular and extracellular concentration of ATP was determined. The bacterial strains were treated with two irradiation doses: 1.2 kGy to induce cell damage and 3.5 kGy to cause cell death for L. monocytogenes. A dose of 0.4 kGy to induce cell damages, 1.1 kGy to obtain viable but nonculturable (VBNC) state and 1.3 kGy to obtain a lethal dose was also applied on E. coli O157H7. Oregano essential oil was used at 0.020% and 0.025% (w/v), which is the minimum inhibitory concentration (MIC) for L. monocytogenes. For E. coli O157H7, a concentration of 0.006% and 0.025% (w/v) which is the ...

2009-07-15

Identification of actively filling sucrose sinks. [Solanum tuberosum; Phaseolus lunatus; Manihot esculenta; Liquidambar styraciflua L. ; Carya illinoinensis

Homology analyses of the protein sequences of chicken liver and rat mammary gland fatty acid synthases were carried out. The amino acid sequences of the chicken and rat enzymes are 67% identical. If conservative substitutions are allowed, 78% of the amino acids are matched. A region of low homologies exists between the functional domains, in particular around amino acid residues 1059-1264 of the chicken enzyme. Homologies between the active sites of chicken and rat and of chicken and yeast enzymes have been analyzed by an alignment method. A high degree of homology exists between the active sites of the chicken and rat enzymes. However, the chicken and yeast enzymes show a lower degree of homology. The DADPH-binding dinucleotide folds of the {beta}-ketoacyl reductase and the enoyl reductase sites were identified by comparison with a known consensus sequence for the DADP- and FAD-binding dinucleotide folds. The active sites of all of the enzymes are primarily in ...

1989-11-01

Differential expression of farnesyl diphosphate synthase gene from Withania somnifera in different chemotypes and in response to elicitors

Certain actively filling plant sucrose sinks such as a seed, a tuber, or a root can be identified by measuring the uridine diphosphate and pyrophosphate-dependent metabolism of sucrose. Sucrolysis in both active and quiescent sucrose sinks was tested and sucrose synthase was found to be the predominant sucrose breakdown activity. Sucrolysis via invertases was low and secondary in both types of sinks. Sucrose synthase activity dropped markedly, greater than fivefold, in quiescent sinks. The test are consistent with the hypothesis that the sucrose filling activity, i.e. the sink strength, of these plant sinks can be measured by testing the uridine diphosphate and pyrophosphate-dependent breakdown of sucrose. Measuring the initial reactions of sucrolysis shows much promise for use in agriculture crop and tree improvement research as a biochemical test for sink strength.

1989-04-01

British Library Electronic Table of Contents (United Kingdom)

Withania somnifera (L.) Dunal (Family, Solanaceae), commonly known as Ashwagandha is one of the most valuable medicinal plants synthesizing large number of pharmacologically active secondary metabolites known as withanolides. Though the plant has been well characterized in terms of phytochemical profiles as well as pharmaceutical activities, not much is known about the genes responsible for biosynthesis of these compounds. In this study, we have characterized a gene encoding farnesyl diphosphate synthase (FPPS; EC 2.5.1.10), a key enzyme in the pathway of biosynthesis of isoprenoids, from W. somnifera. The full-length cDNA of Withania somnifera FPPS (WsFPPS) of 1,253?bps encodes a polypeptide of 343 amino acids. The amino acid sequence homology and phylogenetic analysis suggest that WsFPPS...

2011-01-01

100

Characterization of the mannan synthase promoter from guar (Cyamopsis tetragonoloba)

British Library Electronic Table of Contents (United Kingdom)

Guar seed gum, consisting primarily of a high molecular weight galactomannan, is the most cost effective natural thickener, having broad applications in the food, cosmetics, paper, pharmaceutical and petroleum industries. The properties of the polymer can potentially be enhanced by genetic modification. Development of suitable endosperm-specific promoters for use in guar is desirable for metabolic engineering of the seed gum. A ~1.6?kb guar mannan synthase (MS) promoter region has been isolated. The MS promoter sequence was fused with the GUS reporter gene and overexpressed in the heterologous species alfalfa (Medicago sativa). The potential strength and specificity of the MS promoter was compared with those of the constitutive 35S promoter and the seed specific ?-phaseolin promoter. Quant...

2011-01-01

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101

c-JUN N-TERMINAL KINASE MODULATES OXIDANT STRESS AND PEROXYNITRITE FORMATION INDEPENDENT OF INDUCIBLE NITRIC OXIDE SYNTHASE IN ACETAMINOPHEN HEPATOTOXICITY

Up-Regulation of the RhoA/Rho-Kinase Signaling Pathway in Corpus Cavernosum from Endothelial Nitric-Oxide Synthase (NOS), but Not Neuronal NOS, Null Mice

Acetaminophen (APAP) overdose, which causes liver injury in animals and humans, activates c-jun N-terminal kinase (JNK). Although it was shown that the JNK inhibitor SP600125 effectively reduced...Full Text Available

2010-07-01

102

Nucleotide sequence and genetic analysis of a 13.1-kilobase-pair Pseudomonas denitrificans DNA fragment containing five cob genes and identification of structural genes encoding Cob(I)alamin adenosyltransferase, cobyric acid synthase, and bifunctional cobinamide kinase-cobinamide phosphate guanylyltransferase.

We tested the hypothesis that the basal release of nitric oxide (NO) from endothelial cells modulates contractile activity in the corpus cavernosum (CC) via inhibition of the RhoA/Rho-kinase signaling...Full Text Available

2010-04-01

103

Insect-Induced Conifer Defense. White Pine Weevil and Methyl Jasmonate Induce Traumatic Resinosis, de Novo Formed Volatile Emissions, and Accumulation of Terpenoid Synthase and Putative Octadecanoid Pathway Transcripts in Sitka Spruce1[w

A 13.1-kb DNA fragment carrying Pseudomonas denitrificans cob genes has been sequenced. The nucleotide sequence and genetic analysis revealed that this fragment contained five different cob genes named...Full Text Available

1991-10-01

104

Expression Levels of Thymidylate Synthase, Thymidylate Phosphorylase and Dihydropyrimidine Dehydrogenase in Head and Neck Squamous Cell Carcinoma: Preliminary Study

Stem-boring insects and methyl jasmonate (MeJA) are thought to induce similar complex chemical and anatomical defenses in conifers. To compare insect- and MeJA-induced terpenoid responses, we analyzed...Full Text Available

2005-01-01

105

Enzymic synthesis of 1-O-(indol-3-ylacetyl)-beta-D-glucose. Purification of the enzyme from Zea mays, and preparation of antibodies to the enzyme.

IntroductionPharyngo-laryngeal tumors classified as T3-4, N0-3, M0, are conventionally treated by mutilating surgery (total (pharyngo)-laryngectomy). Neo-adjuvant chemotherapy with...Full Text Available

106

Both the Fas Ligand and Inducible Nitric Oxide Synthase Are Needed for Control of Parasite Replication within Lesions in Mice Infected with Leishmania major whereas the Contribution of Tumor Necrosis Factor Is Minimal

The enzyme indol-3-ylacetylglucose synthase (UDP-glucose:indol-3-ylacetate beta-D-glucosyltransferase) catalyses the reaction: [formula: see text] This is the first step in the series of reactions leading...Full Text Available

1991-10-15

107

Serine-324 of myosin's heavy chain is photoaffinity-labeled by 3 prime (2 prime )-O-(4-benzoylbenzoyl)adenosine triphosphate

Following infection with the protozoan parasite Leishmania major, C57BL/6 mice develop a small lesion that heals spontaneously. Resistance to infection is associated with the development...Full Text Available

2003-09-01

108

Helminthosporium maydis T toxin decreased calcium transport into mitochondria of susceptible corn

A portion of the active site of rabbit skeletal myosin near the ribose ring of ATP can be labeled by the photoaffinity analogue 3{prime}(2{prime})-O-(4-benzoylbenzoyl)adenosine triphosphate (Bz{sub 2}ATP). The specificity of the photolabeling was assured by first trapping ({sup 14}C)Bz{sub 2}ATP at the active site by use of thiol cross-linking agents. Five radioactive peptides were isolated by high-performance liquid chromatography after extensive trypsin and subtilisin digestion of photolabeled myosin subfragment 1. Four of these peptides were sequenced by Edman techniques, and all originated from a region with the sequence Gly-Glu-Ile-Thr-Val-Pro-Ser-Ile-Asp-Asp-Gln, which corresponds to rabbit myosin heavy chain residues 312-328. The fifth labeled peptide had an amino acid composition appropriate for residues 312-328. Amino acid composition, radiochemical analysis, and sequence data indicate that Ser-324 is the major ...

1989-05-02

109

Motor Proteins Have Highly Correlated Brownian Engines

The effects of purified Helminthosporium maydis T (HmT) toxin on active Ca/sup 2 +/ transport into isolated mitochondria and microsomal vesicles were compared for a susceptible (T) and a resistant (N) strain of corn (Zea mays). ATP, malate, NADH, or succinate could drive /sup 45/Ca/sup 2 +/ transport into mitochondria of corn roots. Ca/sup 2 +/ uptake was dependent on the proton electrochemical gradient generated by the redox substrates or the reversible ATP synthetase, as oligomycin inhibited ATP-driven CA/sup 2 +/ uptake while KCN inhibited transport driven by the redox substrates. Purified native HmT toxin completely inhibited Ca/sup 2 +/ transport into T mitochondria at 5 to 10 nanograms per milliliter while transport into N mitochondria was decreased slightly by 100 nanograms per milliliter toxin. Malate-driven Ca/sup 2 +/ transport in T mitochondria was frequently more inhibited by 5 nanograms per milliliter toxin ...

1984-04-01

110

CERN Document Server

Two headed motor proteins, such as kinesin and dynein, hidrolyze environmental ATP in order to propel unidirectionally along cytoskeletal filaments such as microtubules. In the case of kinesin, protein heads bind primarily on the alpha tubulin site of asymmetric alpha-beta 8nm-long tubulin dimers that constitute the microtubular protofilaments. Kinesin dimers overcome local binding forces up to 5pN and are known to move on protofilaments with ATP concentration-dependent speeds while hydrolizing on average one ATP molecule per 8nm step. The salient features of protein trajectories are the distinct abrupt usually 8nm-long steps from one tubulin dimer to the next interlaced with long quiescent binding periods at a tubulin site. Discrete walks of this type are characterized by substantially reduced variances compared to pure biased random walks, and as a result rule out flashing-type ratchet models as possible mechanisms for ...

1998-01-01

111

Hydrolysis of adenosine 5'-triphosphate: an isotope-labeling study

Retrospective Monte Carlo dose calculations with limited beam weight information

A combination of /sup 18/O-labeling experiments and kinetic studies to clarify the nonenzymatic hydrolytic pathways of adenosine 5'-triphosphate (ATP) at pH values ranging from 0 to 8.3 has been used in this experiment. In 1 N and 0.1 N HCl, the data are consistent with the hypothesis that hydrolysis occurs by addition-elimination, with initial attack 93% ..gamma.. and 7% ..beta..; both lead only to ADP + P/sub i/. In the subsequent hydrolysis of the ADP to AMP + P/sub i/, attack is 83% ..beta.. and 17% ..cap alpha... At pH 8.3, the data are consistent with the hypothesis that hydrolsysis occurs by elimination-addition. Over the entire pH range studied, no oxygen exchange between water and ATP, ADP, or P/sub i/ was detected. Nonenzymatic hydrolysis and isotopic analysis of the resultant P/sub i/ comprise a preferred means of assaying the isotopic enrichment of (..gamma..-/sup 18/O)ATP to be used in studies of ...

1982-12-15

112

An important unresolved issue in outcomes analysis for lung complications is the effect of poor or completely lacking heterogeneity corrections in previously archived treatment plans. To estimate this effect, we developed a novel method based on Monte Carlo (MC) dose calculations which can be applied retrospectively to RTOG/AAPM-style archived treatment plans (ATP). We applied this method to 218 archived nonsmall cell lung cancer lung treatment plans that were originally calculated either without heterogeneity corrections or with primitive corrections. To retrospectively specify beam weights and wedges, beams were broken into Monte Carlo-generated beamlets, simulated using the VMC++ code, and mathematical optimization was used to match the archived water-based dose distributions. The derived beam weights (and any wedge effects) were then applied to Monte Carlo beamlets regenerated based on the patient computed tomography densities. Validation of the process was ...

2007-01-01

113

Structural features underlying selective inhibition of protein kinase CK2 by ATP site-directed tetrabromo-2-benzotriazole

Previous studies using in vitro cell culture systems have shown the role of the dynamin-related GTPase Opa1 in apoptosis prevention and mitochondrial DNA (mtDNA) maintenance. However, it remains to...Full Text Available

2011-07-01

114

Selective stimulation of catecholamine release from bovine adrenal chromaffin cells by an ionotropic purinergic receptor sensitive to 2-methylthio ATP

Two novel crystal structures of Zea mays protein kinase CK2α catalytic subunit, one in complex with the specific inhibitor 4,5,6,7-tetrabromobenzotriazole (TBB) and another...Full Text Available

2001-11-01

115

Purinergic receptors are involved in tooth-pulp evoked nocifensive behavior and brainstem neuronal activity

Background2-Methylthioadenosine 5'-triphosphate (2-MeSATP), formerly regarded as a specific P2Y (metabotropic) purinergic receptor agonist, stimulates Ca²⁺influx and...Full Text Available

116

P2X1 receptor mobility and trafficking; regulation by receptor insertion and activation

BackgroundTo evaluate whether P2X receptors are involved in responses to noxious pulp stimulation, the P2X₃and P2X_2/3receptor agonist α,β-methyleneATP...Full Text Available

117

NMR of a synthetic peptide spanning the triphosphate binding site of adenosine 5'-triphosphate in actin

P2X1 receptors for ATP contribute to signalling in a variety of cell types and following stimulation undergo rapid desensitisation (within 1 s), and require ∼5 min to recover. In HEK293 cells...Full Text Available

2010-06-01

118

Interaction of N-methyl-2-alkenyl-4-quinolones with ATP-dependent MurE ligase of Mycobacterium tuberculosis: antibacterial activity, molecular docking and inhibition kinetics

The amino acid residues 114-118 in actin were found to be implicated strongly in the binding of nucleotide, and as would be expected for such an important binding site, they are located in a completely conserved region of the actin sequence. A 19-residue peptide with the actin sequence 106-124 was synthesized in order to span the putative triphosphate binding site. Proton NMR spectra of the actin peptide 114-118 in the presence and absence of ATP indicated that Arg-116 and Lys-118 are particularly involved in binding ATP. A strong binding of ATP to the peptide 106-124 also was measured. Tripolyphosphate bound to the peptide 106-124 somewhat more weakly than ATP. Binding involved residues 115-118 and 121-124, indicating the presence of a reverse turn between these segments. Proton resonances were assigned by using two-dimensional double quantum correlated spectroscopy, one-dimensional spin decoupling ...

1987-03-10

119

Identification of a nucleoside triphosphate binding site on calf thymus RNA polymerase II

ObjectivesThe aim of this study was to comprehensively evaluate the antibacterial activity and MurE inhibition of a set of N-methyl-2-alkenyl-4-quinolones found...Full Text Available

2011-08-01

120

Expression of Genes Encoding F₁-ATPase Results in Uncoupling of Glycolysis from Biomass Production in Lactococcus lactis

A nucleoside triphosphate binding site on calf thymus RNA polymerase II was identified by using photoaffinity analogues of adenosine 5'-triphosphate and guanosine 5'-triphosphate. Both radiolabeled 8-azidoadenosine 5'-triphosphate (8-N3ATP) and radiolabeled 8-azidoguanosine 5'-triphosphate (8-N3GTP) bound to a single polypeptide of this enzyme. This polypeptide has a molecular mass of 37 kilodaltons and an isoelectric point of 5.4. Ultraviolet (UV) irradiation was necessary for photolabeling to occur. In addition, no labeling occurred when the probe was prephotolyzed or when the enzyme was inactivated. Furthermore, photolabeling of the enzyme could be decreased by preincubation with natural substrates. To provide evidence that the radiolabeled polypeptide forms a part of the domain of the nucleoside triphosphate binding site, experiments were performed using unlabeled 8-N3ATP. Although this unlabeled analogue ...

1986-01-14

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121

Evolution of a bifunctional enzyme: 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase.

We studied how the introduction of an additional ATP-consuming reaction affects the metabolic fluxes in Lactococcus lactis. Genes encoding the hydrolytic part of the F₁ domain...Full Text Available

2002-09-01

122

Disruption of the ATP-binding Cassette B7 (ABTM-1/ABCB7) Induces Oxidative Stress and Premature Cell Death in Caenorhabditis elegans*

The bifunctional rat liver enzyme 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase (ATP:D-fructose-6-phosphate 2-phosphotransferase/D-fructose-2,6-bisphosphate 2-phosphohydrolase, EC 2.7.1.105/EC...Full Text Available

1989-12-01

123

Characterization of the major phosphoprotein and its kinase on the surface of the rat adipocyte

X-linked sideroblastic anemia with ataxia (XLSA/A) is a rare inherited disorder characterized by mild anemia and ataxia. XLSA/A is caused by mutations in the ABCB7 gene, which encodes...Full Text Available

2011-06-17

124

Changes in the profile structure of the sarcoplasmic reticulum membrane induced by phosphorylation of the Ca2+ ATPase enzyme in the presence of terbium: a time-resolved x-ray diffraction study.

Intact rat fat cell exposed to 12.5 ..mu..M (..gamma..-32P)ATP incorporate label into specific proteins within minutes. By solubilizing the reaction mixture with SDS which bypasses the subcellular fractionation steps, the labeled proteins can be identified in autoradiographs of SDS-PAGE gels. The most prominently labeled protein has an M/sub r/ of 42,000. Localization of this component to the cell surface can be made on the basis of inhibition of phosphorylation by addition of a protein derived from the rat brain with protein kinase inhibitory property, susceptibility of the phosphorylated protein to the tryptic digestion, inhibition of phosphorylation of this protein after brief exposure to melittin. To rule out the possibility that the cell surface protein might be a mitochondrial contaminant from broken cells, /sup 32/Pi-labeled and (..gamma..-/sup 32/P)ATP-labeled cells were chromatographed on a rabbit anti-pyruvate dehydrogenase ...

1986-12-01

125

Autocrine regulation of T-cell activation by ATP release and P2X₇ receptors

The design of the time-resolved x-ray diffraction experiments reported in this and an accompanying paper was based on direct measurements of enzyme phosphorylation using [gamma-32P]ATP that were employed...Full Text Available

1994-05-01

126

Thymidylate synthase expression and genotype have no major impact on the clinical outcome of colorectal cancer patients treated with 5-fluorouracil

T-cell activation requires the influx of extracellular calcium, although mechanistic details regarding such activation are not fully defined. Here, we show that P2X₇ receptors play a key...Full Text Available

2009-06-01

127

British Library Electronic Table of Contents (United Kingdom)

Background and objectives.Thymidylate synthase (TS) expression levels appear to be related to response to 5-fluorouracil-(5-FU)-based chemotherapy in colorectal cancer (CRC) patients. Three polymorphisms have been proposed as modulators of TS expression: a tandemly repeated sequence (2R/3R) in the 5? UTR, a SNP (G > C) within the 3R allele and a 6 bp deletion in the 3' UTR.To evaluate the influence of TS expression and polymorphisms on clinical outcome of 5-FU-treated patients we performed a comprehensive genetic analysis on 63 CRC patients.Methods.TS expression levels were analyzed in normal and tumor tissues. TS coding sequence and UTR polymorphisms were investigated on DNA from normal tissue. LOH analysis was performed to determine tumor genotype.Results.A difference in disease-free sur...

2011-01-01

128

The Structure of Sucrose Phosphate Synthase from Halothermothrix orenii Reveals Its Mechanism of Action and Binding Mode

Sucrose phosphate synthase activity in rice grown at elevated CO sub 2 and temperature

Sucrose phosphate synthase (SPS) catalyzes the transfer of a glycosyl group from an activated donor sugar, such as uridine diphosphate glucose (UDP-Glc), to a saccharide acceptor D-fructose 6-phosphate (F6P), resulting in the formation of UDP and D-sucrose-6'-phosphate (S6P). This is a central regulatory process in the production of sucrose in plants, cyanobacteria, and proteobacteria. Here, we report the crystal structure of SPS from the nonphotosynthetic bacterium Halothermothrix orenii and its complexes with the substrate F6P and the product S6P. SPS has two distinct Rossmann-fold domains with a large substrate binding cleft at the interdomain interface. Structures of two complexes show that both the substrate F6P and the product S6P bind to the A-domain of SPS. Based on comparative analysis of the SPS structure with other related enzymes, the donor substrate, nucleotide diphosphate glucose, binds to the B-domain of SPS. Furthermore, we propose a ...

2008-01-01

129

Crystal structures of two novel sulfonylurea herbicides in complex with Arabidopsis thaliana acetohydroxyacid synthase

Rice (Oryza sativa L.) was grown at 330 and 660 {mu}L CO{sub 2} L{sup {minus}1} and at 40/33/37, 34/27/31, and 28/21/25{degree}C day/night/paddy water temperatures respectively. Sucrose phosphate synthase (SPS) activity was measured at saturating substrate concentrations at 59 days after planting. SPS activity increased 2 and 3 fold with increasing CO{sub 2} at 28 and 34{degree}C air temperatures respectively. At 40{degree}C SPS activity decreased by 37% at elevated CO{sub 2} and most plants failed to reach maturity. Similar responses were found in leaf samples taken in the dark. These results indicate that SPS, an enzyme involved in the regulation of C partitioning in leaves, increases in activity at elevated CO{sub 2}. This is in contrast to previous results with soybean. The changes in SPS activity will also be discussed in relation to leaf starch/sucrose ratios.

1990-05-01

130

A rapid, radiometric assay for sucrose synthase

Acetohydroxyacid synthase (AHAS; EC 2.2.1.6) is the first enzyme in the biosynthetic pathway of the branched-chain amino acids. It catalyzes the conversion of two molecules of pyruvate into 2-acetolactate or one molecule of pyruvate and one molecule of 2-ketobutyrate into 2-aceto-2-hydroxybutyrate. AHAS requires the cofactors thiamine diphosphate (ThDP), Mg{sup 2+} and FAD for activity. The herbicides that target this enzyme are effective in protecting a broad range of crops from weed species. However, resistance in the field is now a serious problem worldwide. To address this, two new sulfonylureas, monosulfuron and monosulfuron ester, have been developed as commercial herbicides in China. These molecules differ from the traditional sulfonylureas in that the heterocyclic ring attached to the nitrogen atom of the sulfonylurea bridge is monosubstituted rather than disubstituted. The structures of these compounds in complex with the catalytic subunit of Arabidopsis ...

2009-08-17

131

Single-molecular analysis of the binding state of myosin V and actin

Investigations of sucrose synthase in maize root tips have required development of a means to circumvent the rapid decline of activity observed after extraction dialysis and either synthetic or degradative assays. Several protease inhibitors were tested; although PMSF increased initial activity, no inhibitor prevented the drop in activity with time. Western blot analysis indicated that activity decline was not associated with protein degradation. Therefore, a procedure was developed which (1) shortened extraction-to-assay period from ca. 24 hours to 7 minutes, (2) simplified previous assays and (3) reduced the amount of tissue required. Extract was desalted with spun columns and the {sup 14}C-UDPG product recovered with DEAE ion exchange paper. The minute quantities of product recovered can be concealed by the presence of trace impurities in the {sup 14}C-sucrose utilized. DEAE ion exchange paper was used to remove interfering radio-labelled compounds from the {sup ...

1990-05-01

132

Rapid toxicity testing based on mitochondrial respiratory activity

A class-V myosin, myosin V, one of 18 known classes of actin-based motor proteins, plays a role in transporting organelles within a cell. Unlike myosin-II, which functions as an assembly in the thick filaments of muscle, myosin V is a two-headed processive motor protein, which functions as a single molecule: myosin V performs many consecutive steps before it detaches from an actin filament accompanied by catalytic cycles of ATP (adenosine 5'-triphosphate) ase. The mechanism of such chemomechanical steps is explained by a 'Hand-over-hand model' in which two heads of myosin V alternately repeat single-headed and double-headed bindings with an actin filament. To investigate the binding state of myosin V at several key nucleotide states during ATP hydrolysis, we measured the mechanical properties of a single myosin V - actin complex by applying an external load with optical trap.

2006-03-21

133

Radioactive-electrophoretic assay of adenosine 5'-triphosphate sulfurylase activity in crude extracts with sulfate or selenate as a substrate

The need exists for rapid and inexpensive methods to determine the health effects of environmental contaminants on biological systems. One of the current research approaches for assessing cytotoxicity is to monitor the respiratory activity of the mitochondrion, a sensitive, nonspecific subcellular target site. Detected changes in mitochondrial function after the addition of a test chemical could be correlated to toxic effects. Mitochondrial respiration can be characterized by three indices: state 3 and state 4 respiratory rates, and the respiratory control ratio (RCR). State 4, the idle or resting state, results when coupled mitochondrial respire in a medium containing inorganic phosphate and a Kreb's cycle substrate in the absence of a phosphate acceptor such as adenosine diphosphate (ADP). In the presence of ADP the respiration rate increases to a maximum (state 3), accompanied by phosphorylation of ADP to adenosine triphosphate (ATP). The ratio of state ...

1990-05-01

134

Effect of pH on the phase separation in the ternary aqueous system containing the hydrophilic ionic liquid 1-butyl-3-methylimidazolium bromide and the kosmotropic salt potassium citrate at T=298.15K

An assay method for ATP sulfurylase is presented which employs Na/sub 2/(35)SO/sub 4/ as a substrate and measures the production of labeled adenosine 5'-phosphosulfate and 3'-phosphoadenosine 5'-phosphosulfate by low-voltage, hanging paper strip electrophoresis. The method is applicable to crude bacterial or mammalian extracts and accurately measures picomole amounts of product(s). Na/sub 2/(/sup 75/)SeO/sub 4/ can also be employed as a substrate, if the unstable radioactive product, adenosine 5'-phosphoselenate, is converted to elemental /sup 75/Se degrees by inclusion of reduced glutathione in the reaction mixture. The same paper strip electrophoretic technique can then be used to separate /sup 75/Se degrees from the radiolabeled substrate. The method also has utility for measuring any direct reduction by crude microbial extracts of radioactive selenate to selenite, independent of ATP sulfurylase.

1989-02-01

135

British Library Electronic Table of Contents (United Kingdom)

The goal of this work is to study the effect of the aqueous medium pH on the two-phase equilibrium behaviour of the aqueous {1-butyl-3-methylimidazolium bromide ([C4mim]Br)+potassium citrate} system in order to obtain further information about the salting-out effect produced by the addition of a kosmotropic salt to an aqueous solution of a hydrophilic ionic liquid (IL). For this purpose the phase diagrams and the liquid-liquid equilibrium (LLE) data of the {[C4mim]Br+potassium citrate} aqueous two-phase system (ATPS) were determined experimentally at different pH values. Four medium pH values (5.00, 6.00, 7.00 and 8.00) were assayed at T=298.15K. The effect of pH on the ATPS-promoting capability of citrate anions in the aqueous [C4mim]Br solutions was explained based on the structural hydr...

2011-01-01

136

Direct photoaffinity labeling by nucleotides of the apparent catalytic site on the heavy chains of smooth muscle and Acanthamoeba myosins

Photoaffinity labeling of ATP and NAD"+ binding sites on recombinant human interleukin 2

The heavy chains of Acanthamoeba myosins, IA, IB and II, turkey gizzard myosin, and rabbit skeletal muscle myosin subfragment-1 were specifically labeled by radioactive ATP, ADP, and UTP, each of which is a substrate or product of myosin ATPase activity, when irradiated with uv light at 0"0C. With UTP, as much as 0.45 mol/mol of Acanthamoeba myosin IA heavy chain and 1 mol/mol of turkey gizzard myosin heavy chain was incorporated. Evidence that the ligands were associated with the catalytic site included the observations that reaction occurred only with nucleotides that are substrates or products of the ATPase activity; that the reaction was blocked by pyrophosphate which is an inhibitor of the ATPase activity; that ATP was bound as ADP; and that label was probably restricted to a single peptide following limited subtilisin proteolysis of labeled Acanthamoeba myosin IA heavy chain and extensive cleavage with CNBr and trypsin of labeled turkey ...

137

Kinetic, spectroscopic and chemical modification study of iron release from transferrin; iron(III) complexation to adenosine triphosphate

Interleukin 2 (IL-2) is a T-cell-derived lymphokine critical in the activation and proliferation of T cells, B cells, and lymphokine-activated killer cells. It is a glycoprotein of #approx#15,500 daltons that is synthesized and secreted after activation by antigen or mitogen. By using the analogs 8-azidoadensoine 5'-[#gamma#-"3"2P]triphosphate ([#gamma#-"3"2P]8N_3ATP) and nicotinamide 2-azidoadenine [adenylate-"3"2P]dinucleotide ([#alpha#-"3"2P]2N_3NAD"+) as photoaffinity probes, the authors have detected specific, metal ion-requiring nucleotide binding sites on recombinant human IL-2 (rhIL-2). The specificity of these nucleotide interactions with rhIL-2 was demonstrated by saturation effects and by competition by the parent nucleotides at physiologically relevant concentrations. Saturation of photoinsertion into rhIL-2 occurred at 50 #mu#M [#gamma#-"3"2P]8N_3ATP. Saturation of photoinsertion with [#alpha#-"3"2P]2N_3NAD"+ was observed at 180 ...

138

Insulin-like growth factor II receptor is phosphorylated by a tyrosine kinase in adipocyte plasma membranes

Amino acids other than those that serve as ligands have been found to influence the chemical properties of transferrin iron. The catalytic ability of pyrophosphate to mediate transferrin iron release to a terminal acceptor is largely quenched by modification non-liganded histine groups on the protein. The first order rate constants of iron release for several partially histidine modified protein samples were measured. A statistical method was employed to establish that one non-liganded histidine per metal binding domain was responsible for the reduction in rate constant. These results imply that the iron mediated chelator, pyrophosphate, binds directly to a histidine residue on the protein during the iron release process. EPR spectroscopic results are consistent with this interpretation. Kinetic and amino acid sequence studies of ovotransferrin and lactoferrin, in addition to human serum transferrin, have allowed the tentative assignment of His-207 in the N-terminal domain and His-535 ...

1985-01-01

139

Expression of Na(+)-H+ exchange and ATP-dependent proton extrusion in growing rat IMCD cells

Incorporation of /sup 32/P from (gamma-32P)ATP into tyrosine residues of the insulin-like growth factor (IGF)-II receptor was observed in a Triton X-100-insoluble fraction of rat adipocyte plasma membranes. IGF-II receptor phosphorylation proceeded to a stoichiometry of approximately 0.5 mol of phosphate/IGF-II binding site after 10 min of incubation at 4 degrees C. A Km for ATP of 6 microM was calculated for this phosphorylation reaction. Addition of IGF-II caused an approximately 2-fold increase in tyrosine phosphorylation of the IGF-II receptor in this preparation. In contrast, phosphorylation of angiotensin II by the Triton X-100 washed membranes was not stimulated by IGF-II. Incubation of purified receptor immobilized on IGF-II agarose or of receptor-enriched low density microsomal membranes with (gamma-32P)ATP did not result in appreciable incorporation of (/sup 32/P)phosphate into the IGF-II receptor nor into ...

1986-06-15

140

ATP-dependent partitioning of the DNA template into supercoiled domains by Escherichia coli UvrAB

As the last step of urinary acidification, the inner medullary collecting duct (IMCD) is thought to secrete protons into the tubular lumens by means of a H(+)-translocating adenosinetriphosphatase (H(+)-ATPase). However, recent studies have also shown the existence of Na(+)-H+ exchange activity in IMCD cells. Although the physiological function of the antiporter in IMCD cells is unknown, activation of Na(+)-H+ exchange in other cell-culture systems has been suggested to be closely associated with the process of cell growth. Thus presence of Na(+)-H+ exchange may relate to the growth phase of these cells. To examine intracellular pH (pHi) regulation in growing IMCD cells, we studied proton transport by Na(+)-dependent and Na(+)-independent mechanisms by microfluorimetry using the pHi-sensitive dye 2',7'-bis(carboxyethyl)-5,6-carboxyfluorescein acetoxymethyl ester (BCECF/AM). Actively growing cells, defined by (3H)thymidine incorporations, demonstrated an ...

1990-03-01

1 2 3 4 5 6 7 8 9 10 11

141

Sucrose synthesizing enzymes and /sup 14/C-assimilation in the chlorophyllous layer of developing grapefruit. [Citrus paradisi Macf

The helicase action of the Escherichia coli UvrAB complex on a covalently closed circular DNA template was monitored using bacterial DNA topoisomerase I, which specifically removes negative supercoils. In the presence of E. coli DNA topoisomerase I and ATP, the UvrAB complex gradually introduced positive supercoils into the input relaxed plasmid DNA template. Positive supercoils were not produced when E. coli DNA topoisomerase I was replaced by eukaryotic DNA topoisomerase I or when both E. coli and eukaryotic DNA topoisomerases I were added simultaneously. These results suggest that like other DNA helix-tracking processes, the ATP-dependent action of the UvrAM complex on duplex DNA simultaneously generates both positive and negative supercoils, which are not constrained by protein binding but are torsionally strained. The supercoiling activity of UvrAB on UV-damaged DNA was also studied using UV-damaged plasmid DNA and a mutant UvrA protein ...

142

Metabolic and molecular stress responses of gilthead seam bream Sparus aurata during exposure to low ambient temperature: an analysis of mechanisms underlying the winter syndrome

Fixation of /sup 14/CO/sub 2/ and activities of sucrose-synthesizing enzymes, sucrose phosphate synthetase (SPS) and sucrose synthase (SS), were assayed in tissues of developing fruit and source leaves from Citrus paradisi Macf. SPS activity of both the outer, chlorophyllous layer of the fruit (flavedo) and source leaves was 10-fold greater than that of the inner, largely non-chlorophyllous layer of the fruit peel (albedo). In contrast, SS activity of the flavedo was 2-fold greater than that of the albedo and 10-fold greater than that of leaves. Fixation of /sup 14/C-photosynthates in isolated tissues (flavedo 2x > albedo) and their redistribution in intact fruit indicated that flavedo functions as both source and sink. Activities of sucrose-synthesizing enzymes were consistent with this dual function.

1986-04-01

143

British Library Electronic Table of Contents (United Kingdom)

The winter syndrome in the gilthead sea bream Sparus aurata indicates that the species is exposed to critically low temperatures in Mediterranean aquaculture in winter. The present study of metabolic patterns and molecular stress responses during cold exposure was carried out to investigate this ?disease?, in light of the recent concept of oxygen and capacity limited thermal tolerance. The metabolic profile of fuel oxidation was examined by determining the activities of the enzymes hexokinase (HK), aldolase (Ald), pyruvate kinase (PK), l-lactate dehydrogenase (l-LDH), citrate synthase (CS), malate dehydrogenase (MDH) and 3-hydroxyacyl CoA dehydrogenase (HOAD) in heart, red and white muscle after exposure to temperatures of 10, 14 and 18?C. Especially, the increase in LDH activity combined ...

2010-01-01

144

Mechanical wounding induces a nitrosative stress by down-regulation of GSNO reductase and an increase in S-nitrosothiols in sunflower (Helianthus annuus) seedlings

British Library Electronic Table of Contents (United Kingdom)

Nitric oxide (NO) and related molecules such as peroxynitrite, S-nitrosoglutathione (GSNO), and nitrotyrosine, among others, are involved in physiological processes as well in the mechanisms of response to stress conditions. In sunflower seedlings exposed to five different adverse environmental conditions (low temperature, mechanical wounding, high light intensity, continuous light, and continuous darkness), key components of the metabolism of reactive nitrogen species (RNS) and reactive oxygen species (ROS), including the enzyme activities L-arginine-dependent nitric oxide synthase (NOS), S-nitrosogluthathione reductase (GSNOR), nitrate reductase (NR), catalase, and superoxide dismutase, the content of lipid hydroperoxide, hydrogen peroxide, S-nitrosothiols (SNOs), the cellular level of N...

2011-01-01

145

Isoflavone content and apoptotic effect in HT-29 cancer cells of a soy germ extract

British Library Electronic Table of Contents (United Kingdom)

Colon cancer is one of the leading causes of death and a major public health problem in western countries. We examined the isoflavone content of 70% ethanol extract of soy germ (SG) and its apoptotic effect in HT-29 colon cancer cells. Our results showed that the major isoflavones of the SG extract were daidzein and genistein, and it effectively induced apoptosis in HT-29 cancer cells. In addition, nuclear factor-kappa B (NF-kB) expression was reduced in cells treated with the SG extract, which reduced cyclooxygenase-2 (COX-2) and nitric oxide synthase (iNOS) expression. These combined effects ultimately resulted in apoptosis via caspase-3 activation. In addition, daidzein and genistein, the major isoflavones of SG extract, also exerted the apoptotic effect against HT-29 cancer cells. Thes...

2012-01-01

146

Effects of Dietary Glycyrrhizin on Growth and Nonspecific Immunity of White Shrimp, Litopenaeus vannamei

British Library Electronic Table of Contents (United Kingdom)

The growth response, total hemocyte count (THC), respiratory burst (release of superoxide anion), phenoloxidase (PO) activity, superoxide dismutase (SOD) activity, nitric oxide synthase (NOS) activity as well as resistance to the pathogen Vibrio alginolyticus were measured in Litopenaeus vannamei, which had been fed diets supplemented with glycyrrhizin (the aqueous extract of licorice, Glycyrrhiza glabra, roots) at 0, 50, 100, 150, and 200 mg/kg of feed for 8 wk. In the feeding trial, there was no significant difference in survival rate (P > 0.05). Significant higher specific growth rate was observed in treatments with dietary glycyrrhizin than that in the control group (P < 0.05). Shrimp fed a diet with 200 mg/kg of glycyrrhizin had significant higher THC, PO activity, respiratory burst a...

2010-01-01

147

Distribution of NADPH-diaphorase activity in the central nervous system of the young and adult land snail Megalobulimus abbreviatus

British Library Electronic Table of Contents (United Kingdom)

Nitric oxide (NO) is a gas produced through the action of nitric oxide synthase that acts as a neurotransmitter in the central nervous system (CNS) of adult gastropod mollusks. There are no known reports of the presence of NOS-containing neurons and glial cells in young and adult Megalobulimus abbreviatus. Therefore, NADPH-d histochemistry was employed to map the nitrergic distribution in the CNS of young and adult snails in an attempt to identify any transient enzymatic activity in the developing CNS. Reaction was observed in neurons and fibers in all CNS ganglia of both age groups, but in the pedal and cerebral ganglia, positive neurons were more intense than in other ganglia, forming clusters symmetrically located in both paired ganglia. However, neuronal NADPH-d activity in the mesocer...

2010-01-01

148

ALK, the Key Gene for Gelatinization Temperature, is a Modifier Gene for Gel Consistency in Rice

British Library Electronic Table of Contents (United Kingdom)

Abstract Gelatinization temperature (GT) is an important parameter in evaluating the cooking and eating quality of rice. Indeed, the phenotype, biochemistry and inheritance of GT have been widely studied in recent times. Previous map-based cloning revealed that GT was controlled by ALK gene, which encodes a putative soluble starch synthase II-3. Complementation vector and RNAi vector were constructed and transformed into Nipponbare mediated by Agrobacterium. Phenotypic and molecular analyses of transgenic lines provided direct evidence for ALK as a key gene for GT. Meanwhile, amylose content, gel consistency and pasting properties were also affected in transgenic lines. Two of four nonsynonymous single nucleotide polymorphisms in coding sequence of ALK were identified as essential for GT. ...

2011-01-01

149

A Hypothesis: Supplementation with Mushroom-Derived Active Compound Modulates Immunity and Increases Survival in Response to Influenza Virus (H1N1) Infection.

Structural definition of the active site and catalytic mechanism of 3,4-dihydroxy-2-butanone 4-phosphate synthase

We hypothesize that the mushroom-derived active compound may be a potential strategy for increasing survival in response to influenza virus (H1N1) infection through the stimulation of host innate immune response. The validity of the hypothesis can be tested by immune response to influenza infection as seen through survival percentage, virus clearance, weight loss, natural killer cell cytotoxicity, Tumor Necrosis Factor-? (TNF-?) and Interferon-gamma (IFN-?) levels, lytic efficiency in the spleens of mice and inducible nitric oxide synthase mRNA expressions in RAW 264.7 murine macrophage cells. The hypothesis may improve people's quality of life, reduce the medical cost of our healthcare system and eliminate people's fears of influenza outbreak. PMID:21660092

2011-03-20

150

Coordinated responses of phytochelatin synthase and metallothionein genes in black mangrove, Avicennia germinans, exposed to cadmium and copper

X-ray crystal structures of L-3,4-dihydroxy-2-butanone-4-phosphate synthase from Magnaporthe grisea are reported for the E-SO{sub 4}{sup 2-}, E-{sub 4}{sup 2-}-Mg{sup 2+}, E-SO{sub 4}{sup 2-}-Mn{sup 2+}, E-SO{sub 4}{sup 2-}-Mn{sup 2+}-glycerol, and E-SO{sub 4}{sup 2-}-Zn{sup 2+} complexes with resolutions that extend to 1.55, 0.98, 1.60, 1.16, and 1.00 {angstrom}, respectively. Active-site residues of the homodimer are fully defined. The structures were used to model the substrate ribulose 5-phosphate in the active site with the phosphate group anchored at the sulfate site and the placement of the ribulose group guided by the glycerol site. The model includes two Mg{sup 2+} cations that bind to the oxygen substituents of the C2, C3, C4, and phosphate groups of the substrate, the side chains of Glu37 and His153, and water molecules. The position of the metal cofactors and the substrate's phosphate group are further stabilized by an extensive hydrogen-bond ...

2010-03-08

151

Coordinated responses of phytochelatin synthase and metallothionein genes in black mangrove, Avicennia germinans, exposed to cadmium and copper

To evaluate the role of phytochelatins and metallothioneins in heavy metal tolerance of black mangrove Avicennia germinans, 3-month-old seedlings were exposed to cadmium or copper for 30 h, under hydroponic conditions. Degenerate Mt2 and PCS primers were synthesized based on amino acid and nucleotide alignment sequences reported for Mt2 and PCS in other plant species found in GenBank. Total RNA was isolated from A. germinans leaves and two partial fragments of metallothionein and phytochelatin synthase genes were isolated. Gene expression was evaluated with reverse transcripatase-polymerase chain reaction (RT-PCR) amplification technique. Temporal analysis showed that low Cd{sup 2+} and Cu{sup 2+} concentrations caused a slight (but not significant) increase in AvMt2 expression after a 16 h exposure time, while AvPCS expression showed a significant increase under the same conditions but only after 4 h. Results strongly suggest that the rapid increase in AvPCS ...

2007-08-01

152

Biosynthesis of sucrose and mannitol as a function of leaf age in celery (Apium graveolens L. )

To evaluate the role of phytochelatins and metallothioneins in heavy metal tolerance of black mangrove Avicennia germinans, 3-month-old seedlings were exposed to cadmium or copper for 30 h, under hydroponic conditions. Degenerate Mt2 and PCS primers were synthesized based on amino acid and nucleotide alignment sequences reported for Mt2 and PCS in other plant species found in GenBank. Total RNA was isolated from A. germinans leaves and two partial fragments of metallothionein and phytochelatin synthase genes were isolated. Gene expression was evaluated with reverse transcripatase-polymerase chain reaction (RT-PCR) amplification technique. Temporal analysis showed that low Cd"2"+ and Cu"2"+ concentrations caused a slight (but not significant) increase in AvMt2 expression after a 16 h exposure time, while AvPCS expression showed a significant increase under the same conditions but only after 4 h. Results strongly suggest that the rapid increase in AvPCS expression ...

2007-08-01

153

Tungsten Effects on Phosphate-Dependent Biochemical Pathways are Species and Liver Cell Line Dependent

In celery (Apium graveolens L.), the two major translocated carbohydrates are sucrose and the acyclic polyol mannitol. Their metabolism, however, is different and their specific functions are uncertain. To compare their roles in carbon partitioning and sink-source transitions, developmental changes in /sup 14/CO/sub 2/ labeling, pool sizes, and key enzyme activities in leaf tissues were examined. The proportion of label in mannitol increased dramatically with leaf maturation whereas that in sucrose remained fairly constant. Mannitol content, however, was high in all leaves and sucrose content increased as leaves developed. Activities of mannose-6-P reductase, cytoplasmic and chloroplastic fructose-1,6-bis-phosphatases, sucrose phosphate synthase, and sucrose synthase increased with leaf maturation and decreased as leaves senesced. Ribulose bisphosphate carboxylase and nonreversible glyceraldehyde-3-P dehydrogenase activities rose as leaves ...

1988-01-01

154

British Library Electronic Table of Contents (United Kingdom)

Tungsten, in the form of tungstate, polymerizes with phosphate, and as extensive polymerization occurs, cellular phosphorylation and dephosphorylation reactions may be disrupted, resulting in negative effects on cellular functions. A series of studies were conducted to evaluate the effect of tungsten on several phosphate-dependent intracellular functions, including energy cycling (ATP), regulation of enzyme activity (cytosolic protein tyrosine kinase [cytPTK] and tyrosine phosphatase), and intracellular secondary messengers (cyclic adenosine monophosphate [cAMP]). Rat noncancerous hepatocyte (Clone-9), rat cancerous hepatocyte (H4IIE), and human cancerous hepatocyte (HepG2) cells were exposed to 1-1000 mg/l tungsten (in the form of sodium tungstate) for 24 h, lysed, and analyzed for the ab...

2010-01-01

155

Some sulfonamide drugs inhibit ATPase activity of heat shock protein 90: investigation by docking simulation and experimental validation

British Library Electronic Table of Contents (United Kingdom)

Eight selected sulfonamide drugs were investigated as inhibitors of heat shock protein 90 (Hsp90). The investigation included simulated docking experiments to fit the selected compounds within the binding pocket of Hsp90. The selected molecules were found to readily fit within the ATP-binding pocket of Hsp90 in low-energy poses. The sulfonamides torsemide, sulfathiazole, and sulfadiazine were found to inhibit the ATPase activity of Hsp90 with IC50 values of 1.0, 2.6, and 1.5 mM, respectively. Our results suggest that these well-established sulfonamides can be good leads for subsequent optimization into potent Hsp90 inhibitors.

2011-01-01

156

Nerve-Evoked Constriction of Rat Tail Veins Is Potentiated and Venous Diameter Is Reduced after Chronic Spinal Cord Transection

British Library Electronic Table of Contents (United Kingdom)

Abstract Despite reduced sympathetic activity below the level of a spinal cord injury (SCI), venoconstriction during autonomic dysreflexia increases venous return to the heart. Here, contractions of isometrically mounted tail veins from rats with spinal transection at T4 performed 8???10 weeks earlier are compared with those from sham-operated rats. After SCI, lumen diameter was reduced by ?30% and the contractions evoked by electrical stimulation of the perivascular axons were larger than control. This augmentation of neurovascular transmission was not associated with enhanced sensitivity to ?-adrenoceptor agonists or to adenosine-5?-triphosphate (ATP) although contractions to depolarization with K+ were larger after SCI. The percentage reduction in nerve-evoked contraction after SCI prod...

2011-01-01

157

Mitochondria-localized NAD biosynthesis by nicotinamide mononucleotide adenylyltransferase in Jerusalem artichoke (Helianthus tuberosus L.) heterotrophic tissues

British Library Electronic Table of Contents (United Kingdom)

Current studies in plants suggest that the content of the coenzyme NAD is variable and potentially important in determining cell fate. In cases that implicate NAD consumption, re-synthesis must occur to maintain dinucleotide pools. Despite information on the pathways involved in NAD synthesis in plants, the existence of a mitochondrial nicotinamide mononucleotide adenylyltransferase (NMNAT) activity which catalyses NAD synthesis from nicotinamide mononucleotide (NMN) and ATP has not been reported. To verify the latter assumed pathway, experiments with purified and bioenergetically active mitochondria prepared from tubers of Jerusalem artichoke (Helianthus tuberosus L.) were performed. To determine whether NAD biosynthesis might occur, NMN was added to Jerusalem artichoke mitochondria (JAM)...

2011-01-01

158

Mechanisms of Reproductive Thermotolerance in Gossypium hirsutum: The Effect of Genotype and Exogenous Calcium Application

British Library Electronic Table of Contents (United Kingdom)

Abstract Although photosynthetic thermotolerance has been investigated extensively in cotton leaves, reports on the biochemical influence of the pistil in promoting fertilization thermostability are limited. To evaluate the effect of temperature, genotype, and exogenous calcium application on fertilization and pistil biochemistry in cotton, thermosensitive (cv. ST4554 B2RF) and thermotolerant (cv. VH260) plants were grown under control (30/20-C) or high-temperature (38/20-C) conditions during flowering, and exogenous CaCl2 was applied to flowers 1-day prior to anthesis. Measured pistil parameters included fertilization efficiency; protein concentration; glutathione reductase, superoxide dismutase (SOD) and NADPH oxidase activities; and ATP and calcium levels. Exogenous calcium had no effec...

2011-01-01

159

Impaired mitochondrial energy metabolism and kinetic properties of cytochrome oxidase following acute aluminium phosphide exposure in rat liver

British Library Electronic Table of Contents (United Kingdom)

The present study was designed with an aim to analyze the effect of acute aluminium phosphide (ALP) exposure (10mg/kg b.wt, intragastrically) on the kinetic characteristics of cytochrome oxidase and energy metabolism in male Wistar rat liver mitochondria. Liver mitochondrial preparations from ALP-treated rats exhibited significant decrease (66%) in the activity of cytochrome oxidase suggesting that there was a decrease in the catalytic efficiency of the active oxidase molecules on ALP treatment. The decreased activity of cytochrome oxidase with altered NADH and succinic dehydrogenase activities might have contributed towards a significant decline in state 3 and state 4 respiration as observed. These alterations in the electron transport chain complexes in turn adversely affected the ATP sy...

2010-01-01

160

Fluorocarbons and cardiac arrhythmia: does difluorodichloromethane (FC 12) inhibit cardiac metabolism

Detrital processing in streams exposed to acidic precipitation in the Central Appalachian Mountains

Certain fluorocarbons, such as difluorodichloromethane (FC 12), depress the cardiovascular system by diminution of all the transmembrane ionic conductances in cardiac tissues. Does FC 12 also inhibit active transport and thus enzymatic activity and cellular energy. We measured phosphocreatine (PC), adenosine triphosphate (ATP) and cyclic adenosine monophosphate (AMPc) in rat hearts. Rats were randomly divided into 4 groups; 2 control groups: one breathing a mixture of oxygen (21%) and nitrogen (79%) (group C) and the other breathing the same mixture but simultaneously perfused with 1 microgram/kg/min. epinephrine (groupe E-C); 2 trial groups T and E-T where nitrogen was replaced by FC 12. The maximal FC 12 concentration of 720 micrograms/ml in arterial blood produced no significant difference in the concentrations of these three metabolites compared with controls.

1986-01-01

1 2 3 4 5 6 7 8 9 10 11

161

Conjugation is essential for the anticholestatic effect of NorUrsodeoxycholic acid in taurolithocholic acid induced cholestasis in rat liver

Continuing high rates of acidic deposition in the eastern United States may lead to long-term effects on stream communities, because sensitive catchments are continuing to lose anions and cations. A two-year study of the effects of pH and associated water chemistry variables on detrital processing in three streams with different bedrock geology in the Monongahela National Forest, West Virginia were investigated. Leaf pack processing rates and macroinvertebrate colonization and microbial biomass (ATP concentration) on the packs in the three stream were compared. It was found that macroinvertebrate and microbial communities differed both among streams that differed in their capacity to buffer the effects of acidic precipitation and among years in the same stream; these differences in biotic communities were not large enough to affect rates of leaf processing between the two years of the study, but they did significantly affect processing rates between acidic and ...

162

British Library Electronic Table of Contents (United Kingdom)

Abstract NorUDCA (24 norursodeoxycholic acid), the C23 homolog of ursodeoxycholic acid (UDCA), showed remarkable therapeutic effects in cholestatic Mdr2 (Abcb4) (multidrug resistance protein 2/ATP binding cassette b4) knockout mice with sclerosing/fibrosing cholangitis. In contrast to UDCA, norUDCA is inefficiently conjugated in human and rodent liver, and conjugation has been discussed as a key step for the anticholestatic action of UDCA in cholestasis. We compared the choleretic, anticholestatic, and antiapoptotic properties of unconjugated and taurine conjugated UDCA (C24) and norUDCA (C23) in isolated perfused rat liver (IPRL) and in natrium/taurocholate cotransporting polypeptide (Ntcp) transfected human hepatoma (HepG2) cells. Taurolithocholic acid (TLCA) was used to induce a predomi...

2010-01-01

163

Analysis of even harmonics generation in an isolated electric power system

British Library Electronic Table of Contents (United Kingdom)

Harmonics bred from loads are mainly odd order because the current waveforms have half-wave symmetry. Since the even harmonics are negligibly small, they generally are not measured in electric power systems. However, even harmonics were measured at a 500/275/154 kV substation in Hokuriku Electric Power Company after a transmission line fault was corrected. The even harmonics caused malfunctions of protective digital relays because the relays used 4th harmonics at the input filter as the automatic supervisory signal. This paper describes mechanisms of generation of the even harmonics by comparing the measured waveforms with the ATP-EMTP simulation results. Analysis of these results has clearly shown that there are three different mechanisms of generation of even harmonics. The first mechani...

2009-01-01

164

A PAC containing the human mitochondrial DNA polymerase gamma gene (POLG) maps to chromosome 15q25

Role of mitochondria in sulfolipid biosynthesis by Euglena chloroplasts

The human mitochondrial DNA (mtDNA) is a closed circular, 16,569-bp double-stranded DNA, encoding 13 genes whose protein products are subunits of the oxidative phosphorylation system required for synthesis of most of the ATP consumed by eukaryotic cells. Point mutations of the mtDNA that cause multi-tissue, loss-of-energy syndromes, called mitochondrial encephalomyopathies (e.g., MERRF and MELAS), have been identified. In addition, large-scale deletions of the human mtDNA have been identified and are the molecular bases for the neonatal and adolescent onset loss-of-energy syndromes Pearson and Kearns-Sayer, respectively. 5 refs., 1 fig.

1997-03-01

165

Response of a non-Hodgkin lymphoma to "6"0Co therapy monitored by 31P MRS in situ

Sulfate activation occurs in Euglena mitochondria the authors now find that the sulfate activating enzymes are absent from Euglena chloroplasts. Cells of mutant W/sub 10/BSmL lacking plastids also lack detectable sulfolipid (SL) when grown on /sup 35/SO/sub 4//sup 2 -/ indicating that SL is absent from the mitochondria and is exclusively in the plastids. Plastids alone will convert /sup 35/S-cysteine to /sup 35/SL in the presence of ATP and Mg/sup 2 +/; light is stimulatory. Under similar conditions, chloroplasts and mitochondria incubated together convert /sup 35/SO/sub 4//sup 2 -/ to plastid-localized /sup 35/SL but either organelle incubated alone fails to effect this conversion. Unlabeled cysteine blocks SL labeling from sulfate in the mixed incubation; since cysteine is formed from sulfate by Euglena mitochrondria, cysteine (and other compounds) may move from the mitochondrion to the chloroplast to provide the sulfo group for SL formation. Although ...

1987-04-01

166

Exploring the structural dynamics of the E. coli chaperonin GroEL using translation-libration-screw crystallographic refinement of intermediate states

High quality "3"1P MR spectra (signal to noise ratio (S/N) approximately 18, 15 min acquisition for each spectrum) were consistently obtained with surface coils over a period of 6-week RT. Both transient and steady state alterations in metabolites in response to RT were found in this case. The transient changes occurred during the first 3 hr immediately after the 3rd fractionated RT, these changes include the transient elevation of the PCr resonance, a decrease in PDE and an increase in intracellular pH. The monitoring showed that the metabolites approached steady state approximately 2 hr after the fractionated radiation intervention, suggesting that in vivo MRS can be useful for studying the dynamics of tumor response to RT such as repair of potential lethal damage, growth delay, and reoxygenation etc. The steady-state MR spectra showed the net response to each intervention and can clinically be useful for predicting and measuring the result of the fractionated RT. In this case study, ...

167

Salicylate Treatment Improves Age-Associated Vascular Endothelial Dysfunction: Potential Role of Nuclear Factor kB and Forkhead Box O Phosphorylation

Large rigid-body domain movements are critical to GroEL-mediated protein folding, especially apical domain elevation and twist associated with the formation of a folding chamber upon binding ATP and co-chaperonin GroES. Here, we have modeled the anisotropic displacements of GroEL domains from various crystallized states, unliganded GroEL, ATP?S-bound, ADP-AlFx/GroES-bound, and ADP/GroES bound, using translation-libration-screw (TLS) analysis. Remarkably, the TLS results show that the inherent motions of unliganded GroEL, a polypeptide-accepting state, are biased along the transition pathway that leads to the folding-active state. In the ADP-AlFx/GroES-bound folding-active state the dynamic modes of the apical domains become reoriented and coupled to the motions of bound GroES. The ADP/GroES complex exhibits these same motions, but they are increased in magnitude, potentially reflecting the decreased stability of the complex after nucleotide ...

2004-08-12

168

British Library Electronic Table of Contents (United Kingdom)

We hypothesized that I kappa B kinase (IKK)-mediated nuclear factor kappa B and forkhead BoxO3a phosphorylation will be associated with age-related endothelial dysfunction. Endothelium-dependent dilation and aortic protein expression/phosphorylation were determined in young and old male B6D2F1 mice and old mice treated with the IKK inhibitor, salicylate. IKK activation was greater in old mice and was associated with greater nitrotyrosine and cytokines. Endothelium-dependent dilation, nitric oxide (NO), and endothelial NO synthase phosphorylation were lower in old mice. Endothelium-dependent dilation and NO bioavailability were restored by a superoxide dismutase mimetic. Nuclear factor kappa B and forkhead BoxO3a phosphorylation were greater in old and were associated with increased express...

2011-01-01

169

Impairments of astrocytes are involved in the D-galactose-induced brain aging

Gene discovery in the Acanthamoeba castellanii genome

Astrocyte dysfunction is implicated in course of various age-related neurodegenerative diseases. Chronic injection of D-galactose can cause a progressive deterioration in learning and memory capacity and serve as an animal model of aging. To investigate the involvement of astrocytes in this model, oxidative stress biomarkers, biochemical and pathological changes of astrocytes were examined in the hippocampus of the rats with six weeks of D-galactose injection. D-galactose-injected rats displayed impaired antioxidant systems, an increase in nitric oxide levels, and a decrease in reduced glutathione levels. Consistently, western blotting and immunostaining of glial fibrillary acidic protein showed extensive activation of astrocytes. Double-immunofluorescent staining further showed activated astrocytes highly expressed inducible nitric oxide synthase. Electron microscopy demonstrated the degeneration of astrocytes, especially in the aggregated area of synapse and ...

2008-05-16

170

Duration-controlled swimming exercise training induces cardiac hypertrophy in mice.

Acanthamoeba castellanii is a free-living amoeba found in soil, freshwater, and marine environments and an important predator of bacteria. Acanthamoeba castellanii is also an opportunistic pathogen of clinical interest, responsible for several distinct diseases in humans. In order to provide a genomic platform for the study of this ubiquitous and important protist, we generated a sequence survey of approximately 0.5 x coverage of the genome. The data predict that A. castellanii exhibits a greater biosynthetic capacity than the free-living Dictyostelium discoideum and the parasite Entamoeba histolytica, providing an explanation for the ability of A. castellanii to inhabit adversity of environments. Alginate lyase may provide access to bacteria within biofilms by breaking down the biofilm matrix, and polyhydroxybutyrate depolymerase may facilitate utilization of the bacterial storage compound polyhydroxybutyrate as a food source. Enzymes for the synthesis and breakdown of cellulose were ...

2005-08-01

171

Antifungal thiopeptide cyclothiazomycin B1 exhibits growth inhibition accompanying morphological changes via binding to fungal cell wall chitin

Exercise training associated with robust conditioning can be useful for the study of molecular mechanisms underlying exercise-induced cardiac hypertrophy. A swimming apparatus is described to control training regimens in terms of duration, load, and frequency of exercise. Mice were submitted to 60- vs 90-min session/day, once vs twice a day, with 2 or 4% of the weight of the mouse or no workload attached to the tail, for 4 vs 6 weeks of exercise training. Blood pressure was unchanged in all groups while resting heart rate decreased in the trained groups (8-18%). Skeletal muscle citrate synthase activity, measured spectrophotometrically, increased (45-58%) only as a result of duration and frequency-controlled exercise training, indicating that endurance conditioning was obtained. In groups which received duration and endurance conditioning, cardiac weight (14-25%) and myocyte dimension (13-20%) increased. The best conditioning protocol to promote physiological ...

2003-11-17

172

British Library Electronic Table of Contents (United Kingdom)

Cyclothiazomycin B1 (CTB1) is an antifungal cyclic thiopeptide isolated from the culture broth of Streptomyces sp. HA 125-40. CTB1 inhibited the growth of several filamentous fungi including plant pathogens along with swelling of hyphae and spores. The antifungal activity of CTB1 was weakened by hyperosmotic conditions, and hyphae treated with CTB1 burst under hypoosmotic conditions, indicating increased cell wall fragility. CTB1-sensitive fungal species contain high levels of cell wall chitin and/or chitosan. Unlike nikkomycin Z, a competitive inhibitor of chitin synthase (CHS), CTB1 did not inhibit CHS activity. Although CTB1 inhibited CHS biosynthesis, the same result was also obtained with a non-specific proteins inhibitor, cycloheximide, which did not reduce cell wall rigidity. These ...

2011-01-01

173

Up-regulation of sucrose metabolizing enzymes in Oncidium goldiana grown under elevated carbon dioxide

T-2 mycotoxin inhibits mitochondrial protein synthesis

Experiments were conducted in controlled growth chambers to evaluate how increase in CO{sub 2} concentration affected sucrose metabolizing enzymes, especially sucrose phosphate synthase (SPS; EC 2.4.1.14) and sucrose synthase (SS; EC 2.4.1.13), as well as carbon metabolism and partitioning in a tropical epiphytic orchid species (Oncidium goldiana). Response of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; EC 4.1.1.39) to elevated CO{sub 2} was determined along with dry mass production, photosynthesis rate, chlorophyll content, total nitrogen and total soluble protein content. After 60 days of growth, there was a 80% and 150% increase in dry mass production in plants grown at 750 and 1100 {mu} l{sup -}1 CO{sub 2}, respectively, compared with those grown at ambient CO{sub 2} (about 370 {mu} l{sup -}1). A similar increase in photosynthesis rate was detected throughout the growth period when measured under growth CO{sub 2} conditions. ...

2001-07-01

174

Study on the variations of molecular structures of some biomolecules induced by free electron laser using FTIR spectroscopy

The authors investigated the effect of T-2 toxin on rat liver mitochondrial protein synthesis. Isolated rat liver mitochondria were supplemented with an S-100 supernatant from rat liver and an external ATP-generating system. An in-vitro assay employing cycloheximide, and inhibitor of cytoplasmic protein synthesis, and chloramphenicol, and inhibitor of mitochondrial protein synthesis, to distinguish mitochondrial protein synthesis from the cytoplasmic process. Amino acid incorporation into mitochondria was dependent on the concentration of mitochondria and was inhibited by chloramphenicol. The rate of uptake of tritium leucine into mitochondrial protein was unaffected by the addition of T-2 toxin and was not a rate-limiting step in incorporation. However, 0.02 micrograms/ml of T-2 toxin decreased the rate of protein synthesis inhibition correlated with the amount of T-2 toxin taken up by the mitochondria. While T-2 toxin is known to inhibit eukaryotic protein ...

1988-01-01

175

Protons and glucose metabolism in shock

In this study, free electron laser (FEL) with selective wavelength was used to induce structure changes of biomolecules, which were characterized by FTIR spectroscopy. For understanding of the interactions between FEL and biomolecules as well as biological tissues, the biomolecules investigated are ATP, ADP, AMP, t-RNA, D-ribose and the complex of SmCl_3-D-ribose. Their FTIR spectra before and after irradiation of FEL show molecular structure variations of the samples after irradiation of FEL, especially the rearrangement of their hydrogen bond networks. Along with the various irradiation wavelengths, irradiation time and molecular structures, the changes after irradiation are different for these molecules. In the FTIR spectra after irradiation, the phenomenon that the bands split into several peaks indicates the existence of several structures, conformations and configurations, which may be prompted by multiple photons process induced by FEL. After irradiation, ...

2007-05-01

176

Phosphorylation of proteins in Clostridium thermohydrosulfuricum

When oxygen is limiting, animals can ferment glucose via several metabolic pathways varying in energetic efficiency and leading to various end products (such as lactate, succinate, or propionate). Because the pH dependence of H/sup +/ production by fermentation is opposite to that by hydrolysis of adenosine triphosphate formed in the fermentation, the total number of moles of protons generated is always two per mole of fermentable substrate. However, two and three times more adenosine triphosphate can be turned over per mole of protons produced in succinate and propionate fermentations, respectively, than in lactate fermentation. At its limit, this advantage would achieve the same balance between H/sup +/ production and H/sup +/ consumption during ATP cycling that is observed in aerobic metabolism, a situation observed in certain alcohol fermentations. Since proton balance during anaerobiosis is clearly adaptable, we consider possible impact and functions of net ...

1983-01-01

177

K"A"T"P channels in mesenchymal stromal stem cells: Strong up-regulation of Kir6.2 subunits upon osteogenic differentiation

Cell extracts of the thermophile Clostridium thermohydrosulfuricum catalyzed the phosphorylation by (..gamma..-/sup 32/P)ATP of several endogenous proteins with M/sub r/s between 13,000 and 100,000. Serine and tyrosine were the main acceptors. Distinct substrate proteins were found in the soluble (e.g., proteins p66, p63, and p53 of M/sub r/s 66,000, 63,000, and 53,000, respectively) and particulate (p76 and p30) fractions, both of which contained protein kinase and phosphatase activity. The soluble fraction suppressed the phosphorylation of particulate proteins and contained a protein kinase inhibitor. Phosphorylation of p53 was promoted by 10..mu..M fructose 1,6-bisphosphate or glucose 1,6-bisphosphate and suppressed by hexose monophosphates, whereas p30 and p13 were suppressed by 5 ..mu..M brain (but not spinach) calmodulin. Polyamines, including the odd polyamines characteristic of thermophiles, modulated the labeling of most of the phosphoproteins. Apart from ...

1986-02-01

178

British Library Electronic Table of Contents (United Kingdom)

The promising use of mesenchymal stromal cells (MSC) in regenerative technologies accounts for necessity of detailed study of their physiology. Proliferation and differentiation of multipotent cells often involve changes in their metabolic state. In the present study, we analyzed the expression of ATP-sensitive potassium (K"A"T"P) channels in MSC and upon in vitro differentiation. K"A"T"P channels are present in many cells and regulate a variety of cellular functions by coupling cell metabolism with membrane potential. Kir6.1, Kir6.2 and SUR2A were expressed in undifferentiated MSC, whereas SUR2B and SUR1 were not detected on cDNA and protein level. Upon adipogenic differentiation Kir6.1 and SUR2A showed a significant reduction of the amount of mRNA by 84% and 95%, respectively, whereas Ki...

2011-01-01

179

Intra-cellular transport of single-headed molecular motors KIF1A

CERN Document Server

Motivated by recent experiments on KIF1A, a representative member of single-headed kinesin motor proteins family, we develop a theoretical model of intra-cellular transport by mutually interacting molecular motors. The model explicitly accounts not only for the hydrolysis of ATP, but also for the ratchet mechanism which is believed to drive each individual KIF1A motor. We study the model by a combination of analytical and numerical techniques. A remarkable feature of this model is that all the parameters in it can be completely evaluated from experimental data. Our results in the dilute limit are in excellent quantitative agreement with the empirical data from single molecule experiments. In the high density regime the predictions of the model also agree qualitatively with the corresponding experimental observations. We derive a phase diagram that shows the influence of hydrolysis and Langmuir kinetics on the collective spatio-temporal organization of the motors. ...

2005-01-01

180

Grain boundary self-diffusion of alloy 800 as affected by sulphur, phosphorous and carbon

Grain boundary self-diffusion of alloy 800 as affected by sulphur, phosphorous and carbon

Using a radioactive tracer method the bulk and grain boundary diffusion of {sup 59}Fe was determined in industrial alloy 800 and melts of alloy 800 with additional P and S in the temperature range 800 to 1000 C. The use of the approximation of Suzuoka was confirmed by autoradiographs. In alloy 800 H the activation energy of grain boundary diffusion of {sup 59}Fe is (209 {+-} 17)kJ/mol. Dissolved elements especially P increase the activation energy of the grain boundary diffusion of Fe by their segregation to the grain boundaries. In addition the influence of the grain boundary diffusion on the growth of creep cavities was investigated in the same materials, and the chemical composition of the creep cavities and grain boundaries were analysed by Auger electron spectroscopy (AES). For alloy 800 + 0.088 wt-%P an enrichment of about 14 at-%P was observed at the grain boundaries. The addition of P clearly enhances the creep strength of alloy 800; this can probably be ...

1999-08-01

1 2 3 4 5 6 7 8 9 10 11

181

Comparison of maintenance energy expenditures and growth yields among several rumen bacteria grown on continuous culture.

Using a radioactive tracer method the bulk and grain boundary diffusion of "5"9Fe was determined in industrial alloy 800 and melts of alloy 800 with additional P and S in the temperature range 800 to 1000 C. The use of the approximation of Suzuoka was confirmed by autoradiographs. In alloy 800 H the activation energy of grain boundary diffusion of "5"9Fe is (209 #+-# 17)kJ/mol. Dissolved elements especially P increase the activation energy of the grain boundary diffusion of Fe by their segregation to the grain boundaries. In addition the influence of the grain boundary diffusion on the growth of creep cavities was investigated in the same materials, and the chemical composition of the creep cavities and grain boundaries were analysed by Auger electron spectroscopy (AES). For alloy 800 + 0.088 wt-%P an enrichment of about 14 at-%P was observed at the grain boundaries. The addition of P clearly enhances the creep strength of alloy 800; this can probably be explained ...

1998-07-06

182

Changes in intracellular Ca2+ levels induced by cytokines and P2 agonists differentially modulate proliferation or commitment with macrophage differentiation in murine hematopoietic cells.

Maintenance energy expenditures were mesured for five rumen bacteria, Selenomonas ruminantium, Butyrivibrio fibrisolvens, Bacteroides ruminicola, Megasphaera elsdenii, and Streptococcus bovis, by using a complex medium with glucose as the carbon source. Large differences (as high as 8.5-fold) in maintenance energy expenditures were seen among these bacteria. The suggestion is made that maintenance requirements could be a significant determinant of bacterial competition in the rumen. Theoretical maximum growth yields, calculated from double reciprocal plots of yield versus dilution rate, were compared to theoretical Y(ATP) values in order to estimate minimum molar adenosine 5'-triphosphate yields from glucose for each bacterium. Results showed that relative yield among the bacteria was growth rate dependent. At high dilution rates, both S. ruminantium and S. bovis produced lactate as their principal fermentation product. At lower dilution rates very little lactate ...

1979-03-01

183

Activation of fat cell adenylate cyclase by protein kinase C

The role of intracellular Ca2+ (Ca2+i) on hematopoiesis was investigated in long term bone marrow cultures using cytokines and agonists of P2 receptors. Cytokines interleukin 3 and granulocyte/macrophage colony stimulator factor promoted a modest increase in Ca2+i concentration ([Ca2+]i) with activation of phospholipase Cgamma, MEK1/2, and Ca2+/calmodulin kinase II. Involvement of protein kinase C was restricted to stimulation with interleukin 3. In addition, these cytokines promoted proliferation (20 times) and an increase in the Gr-1(-)Mac-1+ population with participation of gap junctions (GJ). Nevertheless ATP, ADP, and UTP promoted a large increase in [Ca2+]i, moderate proliferation (6 times), a reduction in the primitive Gr-1(-)Mac-1(-)c-Kit+ population, and differentiation into macrophages without participation of GJ. It is likely that Ca2+i participates as a regulator of hematopoietic signaling: moderate increases in [Ca2+]i would be related to ...

2008-09-05

184

A procedure for batch separation of sup 14 C-hexose from sup 14 C-sucrose

Purified protein kinase C (C-kinase) from guinea pig pancreas and rat brain stimulated adenylate cyclase activity in purified rat adipocyte membranes. Cyclase stimulation occurred over 100 to 1000 mU/ml of C-kinase activity, required greater than 10 ..mu..M calcium, proceeded without a lag, was not readily reversible, and required no exogenous phospholipid. Moreover, C-kinase inhibitors, such as chlorpromazine and palmitoyl carnitine, inhibited selectively adenylate cyclase which was activated by C-kinase and calcium. Depending on assay conditions, 10 nM 12-0-tetradecanoylphorbol-13-acetate (TPA) either enhanced or was required for kinase action on cyclase. Also, TPA plus calcium promoted the quantitative association of C-kinase with membranes. Adenylate cyclase activation by C-kinase was seen both in the presence and absence of exogenous GTP, indicating that the kinase effect does not result from an action on the GTP-binding, inhibitory regulatory component (N/sub i/) of the cyclase ...

1986-05-01

185

The genetic basis of salinity tolerance in Arctic charr (Salvelinus alpinus).

This presentation describes a method for separating {sup 14}C-hexose from {sup 14}C-sucrose in extracts of plant tissue. Portions of ethanol extracts are treated with activated charcoal in microcentrifuge tubes. Aliquots are removed, ethanol evaporated and replaced with reaction mixture that phosphorylates hexose (HEPPS, K{sub 2}HPO{sub 4}, Mg(C{sub 2}H{sub 3}O{sub 2}){sub 2}, ovalbumen, Na{sub 2}ATP, yeast hexokinase). After a time course, the hexokinase reaction is stopped (slowed considerably) to minimize effects of contamination enzyme activities. The stopping agent used is lyxose, a nonphosphorylable analogue of glucose. The strong anionic charge of phosphate introduced through the hexokinase action results in binding (> 95%) of hexose-phosphate to anion-exchange resin. Sucrose remains unbound (> 95%) in solution. This batch ion-exchange is performed in microcentrifuge tubes to allow many samples to be processed simultaneously. Recovery of radiolabel in ...

1991-05-01

186

The effect of elevated carbon dioxide on a Sierra-Nevadan dominant species: Pinus ponderosa

ABSTRACT: BACKGROUND: The capacity to maintain internal ion homeostasis amidst changing conditions is particularly important for teleost fishes whose reproductive cycle is dependent upon movement from freshwater to seawater. Although the physiology of seawater osmoregulation in mitochondria-rich cells of fish gill epithelium is well understood, less is known about the underlying causes of inter- and intraspecific variation in salinity tolerance. We used a genome-scan approach in Arctic charr (Salvelinus alpinus) to map quantitative trait loci (QTL) correlated with variation in four salinity tolerance performance traits and six body size traits. Comparative genomics approaches allowed us to infer whether allelic variation at candidate gene loci (e.g., ATP1alpha1b, NKCC1, CFTR, and cldn10e) could have underlain observed variation. RESULTS: Combined parental analyses yielded genome-wide significant QTL on linkage groups 8, 14 and 20 for salinity tolerance performance ...

2011-09-21

187

Seasonal variation of defense-related gene expression in leaves from Bois noir affected and recovered grapevines.

The impact of increasing atmospheric C0{sub 2} has not been fully evaluated on western coniferous forest species. Two year old seedlings of Pinusponderosa were grown in environmentally controlled chambers under increased C0{sub 2} conditions for 6 months. These trees exhibit morphological, physiological, and biochemical alterations when compared to our controls. Analysis of whole plant biomass distribution has shown no significant effect to the root to shoot ratios, however needles subjected to elevated C0{sub 2} exhibited an increased overall specific needle mass and a decreased total needle area. Morphological changes at the needle level included decreased mesophyll to vascular tissue 91 ratio and variations in starch storage in chloroplasts. The elevated CO{sub 2} increased internal CO{sub 2} concentrations and assimilation of carbon. Biochemical assays revealed that ribulose-bisphosphate carboxylase specific activities increased on per unit area basis with C0{sub 2} treatment ...

1995-01-01

188

Regulation of sucrose synthesis in water stressed leaves

Although Bois noir is one of the main phytoplasma diseases of grapevine, the gene expression and enzyme activities that underlie physiological changes occurring in symptomatic and recovered (with spontaneous or induced symptom remission) plants are mostly unknown. Bois noir symptomatic leaves (September 2006, 2007) and symptomless leaves from infected symptomatic plants (September 2007) of Sangiovese (moderately susceptible) and Chardonnay (highly susceptible) cultivars were collected. Moreover, leaves from infected symptomless plants of both cultivars were harvested in June 2007. Leaves from recovered plants were also collected in the same periods. In recovered plants of both cultivars, class III chitinase and almost every time phenylalanine ammonia-lyase and chalcone synthase expression were increased for all collection periods. In symptomatic leaves of both cultivars, the expressions of the same genes were up-regulated and also those of ?-1,3-glucanase and ...

2011-05-31

189

Gene expression profiles deciphering rice phenotypic variation between Nipponbare (Japonica) and 93-11 (Indica) during oxidative stress.

Alteration in carbon metabolism and carbohydrate partitioning occur in drought stressed plants. Some species accumulate large quantities of starch in the chloroplast, which may be used to support sucrose synthesis under conditions of limited carbon supply. The authors monitored chemical partitioning of carbon between sugars and starch and the activity of sucrose phosphate synthase (SPS) and fructose 1,6 bisphosphatase (FBPase) in the source leaves of water stressed tomatoes. Plants were stressed by withdrawing water for 10 days and rewatered for recovery. Water potential dropped from {minus}0.8 to {minus}2.2MPA in 10 days, but recovered to control level 2 days after rewatering. Photosynthetic rates as measured by the activity of Rubisco followed similar patterns to those of water potential. After 10 days, leaf starch levels decreased to less than 50% of control. Sucrose levels did not increase significantly, but hexose levels increased 3-4 fold during the stress ...

1991-05-01

190

Antioxidant treatment with quercetin ameliorates erectile dysfunction in streptozotocin-induced diabetic rats.

Rice is a very important food staple that feeds more than half the world's population. Two major Asian cultivated rice (Oryza sativa L.) subspecies, japonica and indica, show significant phenotypic variation in their stress responses. However, the molecular mechanisms underlying this phenotypic variation are still largely unknown. A common link among different stresses is that they produce an oxidative burst and result in an increase of reactive oxygen species (ROS). In this study, methyl viologen (MV) as a ROS agent was applied to investigate the rice oxidative stress response. We observed that 93-11 (indica) seedlings exhibited leaf senescence with severe lesions under MV treatment compared to Nipponbare (japonica). Whole-genome microarray experiments were conducted, and 1,062 probe sets were identified with gene expression level polymorphisms between the two rice cultivars in addition to differential expression under MV treatment, which were assigned as Core Intersectional Probesets ...

2010-01-08

191

The effect of phytic acid on the levels of blood glucose and some enzymes of carbohydrate and lipid metabolism.

Oxidative stress is demonstrated to be involved in the pathophysiological mechanism of erectile dysfunction (ED). Quercetin, a potent bioflavonoid, has been reported to have the antioxidant role. In the present study, we examined the effect of quercetin on ED and oxidative stress in streptozotocin (STZ)-induced diabetic rats. Diabetes was induced in Sprague-Dawley rats with a single intravenous injection of STZ. The diabetic rats were then randomized to diabetic group and quercetin therapy groups which were treated with quercetin at different doses of 5, 20 and 50mg/kg per day respectively. At the end of the 8th week, erectile function was assessed by measuring the rise in intracavernous pressure (ICP) following cavernous nerve electrostimulation. Superoxide dismutase (SOD) activity, thiobarbituric acid-reacting substance (TBARS) and nitrite and nitrate (NOx) levels were measured in cavernosum tissue. Endothelial NO synthase (eNOS) expression was determined using ...

2011-06-14

192

Small-molecule screen identifies inhibitors of a human intestinal calcium-activated chloride channel.

In this study, six groups of rats were fed as follows: Groups 1 and 2 were fed formulated diets supplemented with zinc or without zinc respectively. Groups 3 and 4 were fed formulated diets supplemented with zinc plus phytic acid extracted from sweet potato (Ipomea batatas) or commercial phytic acid respectively. Groups 5 and 6 were fed formulated diets supplemented with phytic acid extract from sweet potato or commercial phytic acid respectively. The animals were fed for three weeks and then sacrificed The activities of key enzymes of carbohydrate and lipid metabolism as well as transaminases in the liver were determined. Blood glucose level was also assessed. Phytic acid extract consumption from sweet potato and commercial phytic acid plus zinc supplement lowered blood glucose levels. There was no significant change in the activity of 6-phosphogluconate dehydrogenase among the groups. Similarly, phytic acid supplementation showed no significant decrease in the activity of pyruvate ...

2005-03-01

193

Response of photosynthesis, starch and sucrose synthesis to increased CO sub 2 and lowered O sub 2

Calcium-activated chloride channels (CaCCs) are widely expressed in mammalian tissues, including intestinal epithelia, where they facilitate fluid secretion. Potent, selective CaCC inhibitors have not been available. We established a high-throughput screen for identification of inhibitors of a human intestinal CaCC based on inhibition of ATP/carbachol-stimulated iodide influx in HT-29 cells after lentiviral infection with the yellow fluorescent halide-sensing protein YFP-H148Q/I152L. Screening of 50,000 diverse, drug-like compounds yielded six classes of putative CaCC inhibitors, two of which, 3-acyl-2-aminothiophenes and 5-aryl-2-aminothiazoles, inhibited by >95% iodide influx in HT-29 cells in response to multiple calcium-elevating agonists, including thapsigargin, without inhibition of calcium elevation, calcium-calmodulin kinase II activation, or cystic fibrosis transmembrane conductance regulator chloride channels. These compounds also inhibited ...

2007-12-14

194

Open test assembly (OTA) shear demonstration testing work/test plan

Increasing photosynthesis rate by raising CO{sub 2} and lowering O{sub 2} usually results in measured rates lower than those expected from the kinetics of rubisco. The failure of leaves to utilize the increased efficiency of rubisco has been attributed to limitations in rates of either starch or sucrose synthesis, which lead to a lowering of Pi and a reduction in the rate of ATP synthesis. In sugar beet leaves, increasing CO{sub 2} from 350 to 600 {mu}L L{sup {minus}1} usually increased photosynthesis only about 10%, compared to the 45% expected, and lowering O{sub 2} from 2 to 21% increased photosynthesis only an additional 20%, compared to the additional 40% expected. In some cases, raising CO{sub 2} increased photosynthesis the full 30% with no further increase occurring upon lowering O{sub 2}. In all cases, rates of starch and sucrose synthesis consistently increased in proportion to increasing photosynthesis rate. Metabolites that are predominantly cytosolic ...

1990-05-01

195

One-step purification of Taq DNA polymerase using nucleotide-mimetic affinity chromatography.

This document describes the development testing phase associated with the OTA Shear activity and defines the controls to be in place throughout the testing. The purpose of the OTA Shear Program was to provide equipment that is needed for the processing of 40 foot long, sodium wetted, irradiated core components previously used in the FFTF reactor to monitor fuel and materials tests. There are currently 15 of these OTA test stalks located in the Test Assembly Conditioning Station (TACS) inerted vault. These need to be dispositioned for a shutdown mission to eliminate this highly activated, high dose inventory prior to turnover to the ERC since they must be handled by remote operations. These would also need to be dispositioned for a restart mission to free up the vault they currently reside in. The waste handling and cleaning equipment in the J33M Cell was designed and built for the handling of reactor components up to the standard 12 foot length. This program will provide the equipment ...

1998-07-16

196

Internalized insulin-receptor complexes are unidirectionally translocated to chloroquine-sensitive degradative sites. Dependence on metabolic energy

The thermostable Thermus aquaticus DNA polymerase (Taq Pol) has been the key factor in transforming the initial PCR method into one with huge impact in molecular biology and biotechnology. Therefore, the development of effective affinity adsorbents for the purification of Taq Pol, as well as other DNA polymerases, attracts the attention of the enzyme manufacturers and the research laboratories. In this report we describe a simple protocol for the purification of Taq Pol from E. coli lysates, leading to enzymes of high specific activity and purity. The protocol is based on a single affinity chromatography step, featuring an immobilized ligand selected from a structure-biased combinatorial library of dNTP-mimetic synthetic ligands. The ligand library was screened for its ability to bind and purify Taq Pol from E. coli lysates. One immobilized ligand (mABSGu) of the general formula X-Trz-Y, bearing 9-aminoethylguanine (AEGu) and aniline-2-sulfonic acid (mABS) linked on the triazine ...

2007-01-01

197

Effects of chronic swimming training on cardiac sarcolemmal function and composition.

Insulin receptors on the surface of isolated rat adipocytes were photoaffinity labeled at 12 degrees C with the iodinated photoreactive insulin analogue, 125I-B2 (2-nitro-4-azidophenylacetyl)-des-PheB1-insulin, and the pathways in the intracellular processing of the labeled receptors were studied at 37 degrees C. During 37 degrees C incubations, the labeled 440-kDa insulin receptors were continuously internalized (as assessed by trypsin inaccessibility) and degraded such that up to 50% of the initially labeled receptors were lost by 120 min. Metabolic poisons (0.125-0.75 mM 2,4-dinitrophenol (DNP) and 1-10 mM NaF), which led to dose-dependent depletion of adipocyte ATP pools, inhibited receptor loss, and caused up to 3-fold increase in intracellular receptor accumulation. This effect was due to inhibition of intracellular receptor degradation, and there was no apparent effect of the metabolic poisons on initial internalization of the receptors. Following maximal ...

1988-04-25

198

Direct photoaffinity labeling of gizzard myosin with ["3H]uridine diphosphate places Glu185 of the heavy chain at the active site

Cardiac contractile function is dependent on the integrity and function of the sarcolemmal membrane. Swimming exercise training is known to increase cardiac contractile performance. The purpose of the present study was to examine whether a swimming exercise program would alter sarcolemmal enzyme activity, ion flux, and composition in rat hearts. After approximately 11 wk of exercise training, cardiac myosin and actomyosin Ca2+-adenosinetriphosphatase (ATPase) activity was significantly higher in exercised rat hearts than in sedentary control rat hearts. Glycogen content was increased in plantaris and gastrocnemius muscles from exercised animals as was succinic dehydrogenase activity in gastrocnemius muscle of exercised rats in comparison to sedentary rat preparations. Sarcolemmal vesicles were isolated from hearts of exercise-trained and control rats. Sarcolemmal Na+-K+-ATPase and K+-p-nitrophenylphosphatase activities, Na+-Ca2+ exchange, and passive Ca2+ binding did not differ between ...

1989-04-01

199

Digitonin abolishes free 2-deoxy-D-glucose accumulation in isolated rat adipocytes

The active site of chicken gizzard myosin was labeled by direct photoaffinity labeling with ["3H]UDP. ["3H] UDP was stably trapped at the active site by addition of vanadate (Vi) and Co"2"+. The extraordinary stability of the myosin.Co2+.[3H]UDP.Vi complex (t1/2 greater than 5 days at 0 degrees C) allowed it to be purified free of extraneous ["3H]UDP before irradiation began. Upon UV irradiation, greater than 60% of the trapped ["3H]UDP was photoincorporated into the active site. Only the 200-kDa heavy chain was labeled, confirming earlier results using ["3H]UTP. Extensive tryptic digestion of photolabeled myosin subfragment 1 followed by high performance liquid chromatography separations and removal of nucleotide phosphates by treatment with alkaline phosphatase allowed two labeled peptides to be isolated. Sequencing of the labeled peptides and radioactive counting showed that Glu185 was the residue labeled. Since UDP is a zero-length cross-linker, Glu185 is located at the ...

200

Chelation of intracellular calcium blocks insulin action in the adipocyte

The hypothesis that accumulation against sizable chemical gradients of free (non-phosphorylated) 2-deoxy-D-glucose (2dGlc) in isolated rat adipocytes results from an intracellular compartmentation of free hexose was investigated. Cells exposed to 20 ..mu..g/ml digitonin for 10' demonstrated an increased plasma membrane permeability indexed by increased L-glucose entry rates and cellular (presumably cytosolic) protein and K/sup +/ loss. Functional integrity of intracellular organelles was indicated by the ability of the cells to support ATP-driven /sup 45/Ca/sup 2 +/-uptake. Equilibrium 3-O-methylglucose (3-O-MG, a non-accumulated hexose) levels were unaffected. These data suggest a specific permeabilizing action of digitonin at the plasma membrane having no effect on intracellular organelles or passively distributed solutes. Upon addition of digitonin, free 2dGlc fell from 66.5 +/- 8.9 to 7.4 +/- 2.3 pmol/10/sup 5/ cells, a value not significantly ...

1986-03-05

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201

Chelation of intracellular calcium blocks insulin action in the adipocyte

The hypothesis that intracellular Ca/sup 2 +/ is an essential component of the intracellular mechanism of insulin action in the adipocyte was evaluated. Cells were loaded with the Ca/sup 2 +/ chelator quin-2, by preincubating them with quin-2 AM, the tetrakis(acetoxymethyl) ester of quin-2. Quin-2 loading inhibited insulin-stimulated glucose transport without affecting basal activity. The ability of insulin to stimulate glucose uptake in quin-2-loaded cells could be partially restored by preincubating cells with buffer supplemented with 1.2 mM CaCl/sub 2/ and the Ca/sup 2 +/ ionophore A23187. These conditions had no effect on basal activity and omission of CaCl/sub 2/ from the buffer prevented the restoration of insulin-stimulated glucose uptake by A23187. Quin-2 loading also inhibited insulin-stimulated glucose oxidation and the ability of insulin to inhibit cAMP-stimulated lipolysis without affecting their basal activities. Incubation of cells with 100 ..mu..M quin-2 or quin-2 AM had ...

1987-02-01

202

Altered protein phosphorylation in sciatic nerve from rats with streptozocin-induced diabetes

The hypothesis that intracellular Ca"2"+ is an essential component of the intracellular mechanism of insulin action in the adipocyte was evaluated. Cells were loaded with the Ca"2"+ chelator quin-2, by preincubating them with quin-2 AM, the tetrakis(acetoxymethyl) ester of quin-2. Quin-2 loading inhibited insulin-stimulated glucose transport without affecting basal activity. The ability of insulin to stimulate glucose uptake in quin-2-loaded cells could be partially restored by preincubating cells with buffer supplemented with 1.2 mM CaCl_2 and the Ca"2"+ ionophore A23187. These conditions had no effect on basal activity and omission of CaCl_2 from the buffer prevented the restoration of insulin-stimulated glucose uptake by A23187. Quin-2 loading also inhibited insulin-stimulated glucose oxidation and the ability of insulin to inhibit cAMP-stimulated lipolysis without affecting their basal activities. Incubation of cells with 100 #mu#M quin-2 or quin-2 AM had no effect on intracellular ...

203

ATRX ADD domain links an atypical histone methylation recognition mechanism to human mental-retardation syndrome

The effect of experimental diabetes on the phosphorylation of proteins in the rat sciatic nerve was studied. Nerves from animals made diabetic with streptozocin were incubated in vitro with (/sup 32/P)orthophosphate and divided into segments from the proximal to the distal end, and proteins from each segment were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The principal labeled species were the major myelin proteins, P0, and the basic proteins. After 6 wk of diabetes, the incorporation of isotope into these proteins rose as a function of distance along the nerve in a proximal to distal direction and was significantly higher at the distal end compared with incorporation into nerves from age-matched controls. The overall level of isotope uptake was similar in nerves from diabetic animals and weight-matched controls. The distribution of /sup 32/P among proteins also differed in diabetic nerve compared with both control groups in that P0 and the small basic ...

1987-11-01

204

Neurotoxicity of low-dose repeatedly intranasal instillation of nano- and submicron-sized ferric oxide particles in mice

ATR-X (alpha-thalassemia/mental retardation, X-linked) syndrome is a human congenital disorder that causes severe intellectual disabilities. Mutations in the ATRX gene, which encodes an ATP-dependent chromatin-remodeler, are responsible for the syndrome. Approximately 50% of the missense mutations in affected persons are clustered in a cysteine-rich domain termed ADD (ATRX-DNMT3-DNMT3L, ADD{sub ATRX}), whose function has remained elusive. Here we identify ADD{sub ATRX} as a previously unknown histone H3-binding module, whose binding is promoted by lysine 9 trimethylation (H3K9me3) but inhibited by lysine 4 trimethylation (H3K4me3). The cocrystal structure of ADD{sub ATRX} bound to H3{sub 1-15}K9me3 peptide reveals an atypical composite H3K9me3-binding pocket, which is distinct from the conventional trimethyllysine-binding aromatic cage. Notably, H3K9me3-pocket mutants and ATR-X syndrome mutants are defective in both H3K9me3 binding and localization at ...

2011-07-19

205

Two and 8-azido photoaffinity probes. 1. Enzymatic synthesis, characterization, and biological properties of 2- and 8-azido photoprobes of 2-5A and photolabeling of 2-5A binding proteins

Olfactory tract has been demonstrated to be an important portal for inhaled solid nanoparticle transportation into the central nervous system (CNS). We have previously demonstrated that intranasally instilled Fe_2O_3 nanoparticles could transport into the CNS via olfactory pathway. In this study, we investigated the neurotoxicity and size effect of repeatedly low-dose (130 #mu#g) intranasal exposure of nano- and submicron-sized Fe_2O_3 particles (21 nm and 280 nm) to mice. The biomarkers of oxidative stress, activity of nitric oxide synthases and release of monoamine neurotransmitter in the brain were studied. Our results showed that significant oxidative stress was induced by the two sizes of Fe_2O_3 particles. The activities of GSH-Px, Cu,Zn-SOD, and cNOS significantly elevated and the total GSH and GSH/GSSG ratio significantly decreased in the olfactory bulb and hippocampus after the nano- and submicron-sized Fe_2O_3 particle treatment (p < 0.05). The ...

2009-01-01

206