Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:1.2.1.13 (glyceraldehyde-3-phosphate dehydrogenase)
6,511 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

We performed experiments to determine whether nitric oxide promoted the formation of intracellular S-nitrosothiol adducts in human neutrophils. At concentrations sufficient to inhibit chemoattractant-induced superoxide anion production, nitric oxide caused a depletion of measurable intracellular glutathione as determined by both the monobromobimane HPLC method and the glutathione reductase recycling assay. The depletion of glutathione could be shown to be due to the formation of intracellular S-nitrosoglutathione as indicated by the ability of sodium borohydride treatment of cytosol to result in the complete recovery of measurable glutathione. The formation of intracellular S-nitrosylated compounds was confirmed by the capacity of cytosol derived from nitric oxide-treated cells to ADP-ribosylate glyceraldehyde-3-phosphate dehydrogenase. Depletion of intracellular glutathione was accompanied by a rapid and concomitant activation of the hexose monophosphate shunt (HMPS) following exposure to nitric oxide. Kinetic studies demonstrated that nitric oxide-dependent activation of the HMPS was reversible and paralleled nitric oxide-induced glutathione depletion. Synthetic preparations of S-nitrosoglutathione shared with nitric oxide the capacity to inhibit superoxide anion production and activate the HMPS. These data suggest that nitric oxide may regulate cellular functions via the formation of intracellular S-nitrosothiol adducts and the activation of the HMPS.
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PMID:Nitric oxide reacts with intracellular glutathione and activates the hexose monophosphate shunt in human neutrophils: evidence for S-nitrosoglutathione as a bioactive intermediary. 817 Sep 69

After exposure of monkey kidney epithelial cells to a reduced concentration of K, a known mitogenic signal, the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (G3PD) is activated by a cytosolic protein whose function appears to be novel. A monospecific antibody was used as an immunoprobe to study the contribution of this G3PD modifier protein (MP) to signal transduction. Raising the extracellular Na concentration as well as lowering the K concentration of the medium increased the amount of MP in cytosol and also activated G3PD. Metabolic labeling of cells followed by preparation of detergent-soluble (cytosolic) and detergent-resistant (cytoskeletal) fractions, immunoprecipitation, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis of radiolabeled immune precipitates suggested that the protein was also associated with cytoskeleton. Depolymerization of the microtubules with colchicine or nocodazole increased cytosolic immunoreactive MP, whereas cytochalasin D had no effect. Taxol, which stabilizes microtubules, blocked the effects of colchicine or nocodazole. When tubulin, actin, and intermediate filament fractions of the cytoskeleton were prepared, blotted, and probed with specific antibodies, MP was found in the tubulin fraction. These observations suggest that MP is associated with the microtubules and can be displaced into the cytosol, wherein it could activate G3PD and thereby stimulate glycolytic production of ATP during mitogenic signal transduction.
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PMID:Glyceraldehyde-3-phosphate dehydrogenase modifier protein is associated with microtubules in kidney epithelial cells. 818 94

An upshift of the growth temperature from 26 to 40 degrees C in the presence of calcium leads to the aggregation of Kluyveromyces marxianus cells and to the formation of flocs. Analysis of cell wall proteins, either by sodium dodecyl sulphate-polyacrylamide gel electrophoresis of extractable mannoproteins or by immunolocalization, revealed an accumulation of a protein with Mr 37 kDa(p37), upon flocculation. Immunological studies confirmed the homology of this protein with the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH). When mRNA isolated from cells growing at 40 degrees C was translated in vitro, a 35 kDa newly labelled protein was synthesized and immunoprecipitation assays showed that this protein is recognized by p37-antiserum, suggesting that the 35 kDa polypeptide might be an unglycosylated precursor for of p37. The results indicated that the presence of this cell wall mannoprotein closely related to GAPDH is dependent on the growth temperature, suggesting its role as adhesin.
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PMID:Flocculation of Kluyveromyces marxianus is induced by a temperature upshift. 821 93

Incubation of glyceraldehyde-3-phosphate dehydrogenase (GAPD) with sodium nitroprusside (SNP) decreased its activity in concentration- and time-dependent fashion in the presence of a thiol compound, with DTT being more effective than GSH. Both forward and backward reactions were effected. Coinciding with this, HgCl2-sensitive labelling of the protein by [32P]NAD+ also increased, indicating the stimulation of ADP-ribosylation. Treatment with SNP of GAPD samples from rabbit muscle, sheep brain and yeast inactivated the dehydrogenase activity of the three, but only the mammalian proteins showed ADP-ribosylation activity. The SNP-modified protein of rabbit muscle GAPD, freed from the reagent by Sephadex filtration showed a concentration-dependent restoration of the dehydrogenase activity on preincubation with DTT and GSH. Such thiol-treated preparations also gave increased ADP-ribosylation activity with DTT, and to a lesser extent with GSH. The SNP-modified protein was unable to catalyze this activity with the native yeast enzyme and native and heat-inactivated muscle enzyme. It was possible to generate the ADP-ribosylation activity in muscle GAPD, by an NO-independent mechanism, on dialysis in Tris buffer under aerobic conditions, and on incubating with NADPH, but not NADH, in muscle and brain, but not yeast, enzymes. The results suggest that the inverse relationship of the dehydrogenase and ADP-ribosylation activities is coincidental but not correlated.
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PMID:Inverse relationship of the dehydrogenase and ADP-ribosylation activities in sodium-nitroprusside-treated glyceraldehyde-3-phosphate dehydrogenase is coincidental. 821 90

Nitric oxide-stimulated modification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) by [adenylate-32P]NAD has been interpreted in recent reports as ADP-ribosylation. Incubations of GAPDH with the NO-releasing agent sodium nitroprusside (SNP) and NAD resulted, however, in essentially equal incorporation of radiolabel from the adenine, phosphate, and nicotinamide moieties to the extent of approximately 0.02 mol of NAD.mol of GAPDH-1. Modification of GAPDH by free adenosine 5'-diphosphoribose (ADP-ribose) was only 10% of that by NAD. Exposure of GAPDH modified by NAD in the presence of SNP to HgCl2, which acts at thiol linkages, released two products. Both contained nicotinamide and adenylate but did not cochromatograph with NAD. GAPDH activity was inhibited by SNP in a dose-dependent manner in the presence of NAD. When inhibition was 80%, with 1 mM SNP and 1 mM dithiothreitol, covalent modification with NAD was < 2%. This result is consistent with the conclusion that inhibition of GAPDH activity by SNP in the presence of NAD is due primarily to active-site nitrosylation, as reported by other workers, and is not due to the minor modification with NAD. These results demonstrate that NO-stimulated modification of GAPDH with NAD is not ADP-ribosylation as previously reported but rather is covalent binding of NAD through a NO-dependent thiol intermediate, possibly providing an example of an unexpected, altered reactivity of a nitrosylated protein.
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PMID:Stimulation by nitric oxide of an NAD linkage to glyceraldehyde-3-phosphate dehydrogenase. 832 4

We recently identified an enzymatically active glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12; GAPDH) as a major protein on the surface of group A streptococci (SDH), which exhibits multiple binding activity to various mammalian proteins. We now report that the SDH molecule also functions as an ADP-ribosylating enzyme, which, in the presence of NAD, is auto-ADP-ribosylated. In a crude cell wall extract of group A streptococci, SDH is the only protein that is ADP-ribosylated. SDH found in the streptococcal cytoplasmic fraction could not be ADP-ribosylated in the presence of NAD. Treatment of ADP-ribosylated SDH with the cytoplasmic fraction removed the ADP-ribose from SDH, suggesting the presence of an ADP-ribosyl hydrolase in the cytoplasmic compartment. The covalent linkage of ADP-ribose to SDH was stable to neutral hydroxylamine, sensitive to HgCl2, and inhibitable by free cysteine, indicating that the modification was at a cysteine residue of SDH. In addition to its auto-ADP-ribosylation activity, purified SDH or streptococcal cell wall extracts were able to transfer the ADP-ribose moiety of NAD specifically to free cysteine, resulting in a true thioglycosidic linkage. Treatment of purified SDH or the crude cell wall extract with sodium nitroprusside, which spontaneously generates nitric oxide, was found to stimulate the ADP-ribosylation of SDH in a time-dependent manner. ADP-ribosylation and nitric oxide treatment inhibited the GAPDH activity of SDH. Since ADP-ribosylation and nitric oxide are involved in signal transduction events, the ADP-ribosylating activity of SDH may enable communication between host and parasite during infection by group A streptococci.
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PMID:Glyceraldehyde-3-phosphate dehydrogenase on the surface of group A streptococci is also an ADP-ribosylating enzyme. 836 77

Cytotoxic action of membrane lipid peroxidation product 4-hydroxynonenal (HNE) is due mainly to its facile reactivity with proteins (Esterbauer, H., Schaur, R. J., and Zollner, H. (1991) Free Radical Biol. Med. 11, 77-80). In the present study, the detailed mechanism of HNE modification of a key enzyme in intermediary metabolism, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), is studied mainly focusing on the formation of HNE-amino acid adducts in the enzyme. When GAPDH (1 mg/ml) was treated with 0-2 mM HNE in sodium phosphate buffer (pH 7.2) for 2 h at 37 degrees C, the enzyme was inactivated by HNE in a concentration-dependent manner. The loss of enzyme activity was associated with the loss of free sulfhydryl groups. Following its reduction with NaBH4, amino acid analysis of the HNE-modified enzyme demonstrated that histidine and lysine residues were also modified. At concentrations lower than 0.5 mM, HNE reacts preferentially with cysteine and lysine residues. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the HNE-modified enzyme suggested the formation of intra- and intermolecular cross-links of the enzyme subunit. The HNE-dependent loss of amino acid residues was accompanied by the generation of protein-linked carbonyl derivatives as assessed by reduction with NaB[3H]H4 and reaction with 2,4-dinitrophenylhydrazine. Thus, the conjugation of all the amino acids appears to involve Michael addition type reactions in which the carbonyl function of HNE would be preserved. The modified histidine residues were quantitatively recovered as the HNE-histidine adduct. However, only 28% of the missing lysine could be accounted for as the HNE-lysine derivative, and only 15.6% of the modified cysteine could be accounted for as the HNE-cysteine thioether derivative. It is proposed that the carbonyl groups of the HNE-derived Michael addition products may undergo secondary reactions with the amino acid groups of lysine residues to yield inter- and intrasubunit cross-links.
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PMID:Covalent attachment of 4-hydroxynonenal to glyceraldehyde-3-phosphate dehydrogenase. A possible involvement of intra- and intermolecular cross-linking reaction. 845 10

Nitric oxide (NO) stimulates the auto-ADP-ribosylation of the glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) which results in the inhibition of enzyme activity. In the present work we show that addition of exogenous NO or IL-1 beta-induced intracellular NO generation cause GAPDH ADP-ribosylation and inhibition of enzyme activity. Incubation of RINm5F cells with sodium nitroprusside (SNP) for 18 h caused a time- and dose-dependent inhibition of GAPDH activity. Half-maximal inhibition of GAPDH activity was observed with 80 microM of the NO donor, with maximal inhibition after roughly 6 h of incubation. In parallel, SNP induced endogenous ADP-ribosylation of GAPDH measured by a decreased incorporation of [32P]ADP-ribose from [32P]NAD+ in the cytosol of the SNP-treated cells. Stimulation of endogenous NO production by inducing the NO synthase by exposure to the cytokine IL-1 beta results in decreased GAPDH activity. IL-1 beta (10(-9) M) inhibited GAPDH activity about 55%, compared with control values. Production of nitrite and inhibition of GAPDH was reversed by the NO synthase inhibitor NG-monomethyl-L-arginine, indicating that endogenous generated NO was the effective molecule. Again, GAPDH inhibition was associated with NO-stimulated endogenous ADP-ribosylation of the enzyme. Western blot analysis of GAPDH excluded degradation of GAPDH by NO. NO-stimulated auto-ADP-ribosylation resulted in inhibition of the glycolytic enzyme GAPDH and may be relevant as a cytotoxic effect of NO. In concert with its inhibitory actions on iron-sulfur enzymes like aconitase and electron transport proteins of the respiratory chain NO may mediate autocytotoxic effect in beta-cells.
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PMID:Exogenous nitric oxide (NO) generation or IL-1 beta-induced intracellular NO production stimulates inhibitory auto-ADP-ribosylation of glyceraldehyde-3-phosphate dehydrogenase in RINm5F cells. 845 67

The molecular mechanisms involved in the degradation of individual cellular proteins are probably unique and characteristic. We have investigated in rat liver the degradation of glyceraldehyde-3-phosphate dehydrogenase, an abundant cytosolic enzyme of the glycolytic pathway. Immunoblot analysis of isolated liver lysosomes from rats treated with lysosomal inhibitors show that this protein is degraded, at least in part, by a lysosomal pathway. This pathway was further investigated by incubating the enzyme with lysosomes in a cell-free system, followed by proteolysis measurements, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of lysosomes, and electron microscopic immunocytochemistry. We postulate that the degradative mechanism of glyceraldehyde-3-phosphate dehydrogenase includes a temperature-dependent lysosomal pathway, different from classical nonspecific macroautophagy. The postulated pathway involves: binding of the enzyme to the lysosomal membrane, entry into the lysosomal matrix, and degradation. This cell-free system, which can also incorporate in vitro synthesized proteins, should allow further advances toward clarifying the complex signals that regulate protein degradation as well as its close interrelationship with protein synthesis.
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PMID:Uptake and degradation of glyceraldehyde-3-phosphate dehydrogenase by rat liver lysosomes. 848

Recent studies have implicated angiotensin II (angiotensin) in the pathogenesis of cardiac hypertrophy and heart failure. Heart failure is associated with alterations in intracellular Ca2+ movements mediated by sarcolemmal (SL) and sarcoplasmic reticular (SR) membranes in cardiac myocytes. As it was suspected that alteration gene expression of proteins responsible for controlling transmembrane Ca2+ fluxes may contribute to loss of Ca2+ homeostasis in failing hearts, we undertook a study of the effect of angiotensin on the expression of some target genes in the myocardium. Specifically, we tested the effect of angiotensin on mRNA abundance of cardiac Ca(2+)-transport genes including SL Na+/Ca2+ exchange (EX), SR ryanodine receptor (RYR), and sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA). The mRNA abundance of target gene was assessed by Northern blot assay in (i) direct hormonal stimulation of cultured isolated neonatal and adult rat myocytes and (ii) adult rat hearts after implantation of osmotic mini-pumps for delivery of hormone. In all experiments, Northern blot data were normalized using cDNA (Glyceraldehyde 3-phosphate dehydrogenase signal, GAPDH) hybridization to RNA samples. The results indicate that the ratios of EX/GAPDH, RYR/GAPDH, and SERCA2/GAPDH signals were decreased by 51.6%, 55.0%, and 49.4% respectively after neonatal cardiac myocytes were treated (24 h) with 10(-7) M angiotensin. These decreases were blocked completely by treatment with angiotensin subtype 1 (AT1) receptor antagonist (losartan), whereas angiotensin subtype 2 (AT2) receptor antagonist (PD123319) treatment had no effect on the angiotensin-mediated decrease in target gene mRNA abundance. In contrast, angiotensin had no effect on EX, RYR nor SERCA2 gene mRNA abundance in cultured adult myocytes. In a separate series of experiments wherein adult male Sprague-Dawley rats were infused with different dose of angiotensin for 3 days via osmotic mini-pump, we did not detect any alterations in mRNA abundance of cardiac EX/GAPDH, RYR/GAPDH or SERCA/GAPDH genes in either left or right ventricular samples. Thus our results indicate that, in neonatal rat myocytes, angiotensin affects SL and SR calcium transport gene expression by direct agonism of AT1-receptors. As the infusion of low and high dose angiotensin did not affect the expression of target genes in adult hearts, we suggest that the mechanisms for transduction of the angiotensin signaling in neonatal and adult myocytes may be different and may depend on the stage of development. We conclude that regulation of myocardial Ca(2+)-transport gene mRNA abundance by angiotensin may differ among neonatal and adult animals. Nonetheless, our finding with respect to neonatal preparation led us to believe that in neonatal myocytes, the mRNA abundance of SL Na+/Ca2+ exchange, SR ryanodine receptor, and SR Ca(2+)-ATPase are all decreased in response to stimulation by angiotensin.
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PMID:Altered mRNA abundance of calcium transport genes in cardiac myocytes induced by angiotensin II. 876 48


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