Glyceraldehyde-3-phosphate dehydrogenase modification

M22. Mohr, S., Stamler, J. S., and Brune, B., Mechanism of covalent modification of glyceraldehyde-3-phosphate dehydrogenase at its active site thiol by nitric oxide, peroxynitrite and related nitrosating agents. FEBS Lett. 348, 223-227 (1994). 

Experiments with glyceraldehyde-3-phosphate dehydrogenases from M. fervidus and P. woesei as well as with different mutant enzymes [29] indicate that the peptide chain is largely protected from covalent modification in the native protein conformation. It would appear that the native conformation protects the weak links in the chain against attacking water molecules by steric hindrance and by restricting the torsional freedom of the chain. 

C. Lind, R. Gerdes, I. Schuppe-Koistinen, and LA. Cotgreave, Studies on the mechanism of oxidative modification of human glyceraldehyde-3-phosphate dehydrogenase by glutathione catalysis by glutaredoxin, Biochem. Biophys. Res. Commun. 247 (1998) 481-486. 

Inactivation of alcohol dehydrogenase from yeast with 14C-labeled [3-(3-bromoacetylpyridinio)-propyl]-adenosine pyrophosphate followed by oxidation showed the presence of 1-carboxymethyl histidine66. After inactivation of the enzyme with labeled [3-(4-bromoacetylpyridinio)-propyl]-adenosine pyrophosphate followed by oxidation, S-carboxymethyl cysteine was identified in the protein. In the case of glyceraldehyde-3-phosphate dehydrogenase, treatment with either coenzyme analogue leads to the modification of the cysteine residue. Treatment with [14C]nicotinamide-5-bromo-4-methylimidazole dinucleotide did not reveal any modified amino-acid-residues. The labeled nicotinamide residue split off during the recovery of the inactivated enzyme. Attempts to synthesize an inactivator labeled with a 14C-acetyl residue did not give satisfactory yields. If the enzyme-coenzyme derivative was treated with tritiated sodium boron hydride, tritium could be introduced (Fig. 22). Studies with... 

Buchczyk DP, Grune T, Sies H, Klotz LO (2003) Modifications of glyceraldehydes-3-phosphate dehydrogenase induced by increasing concentrations of peroxynitrite early recognition by the proteasome. Biol Chem 384 237-241... 

The administration of TNT to laboratory animals leads to the excretion of 4-NHOH-DNT, 2-NH2-DNT, and 4-NH2-DNT in the urine [59], and to the formation of covalent adducts with microsomal liver and kidney proteins, hemoglobin, and other blood proteins [60], The acid hydrolysis of adducts yielded mainly 2-NH2-DNT (2-ADNT) and 4-NH2-DNT (4-ADNT). Incubation of rat liver microsomes with TNT and NADPH under aerobic conditions resulted in the formation of NH2-DNTs and the transient metabolite 4-NHOH-DNT [57], The formation of covalent protein adducts with TNT metabolites was enhanced by the presence of 02 and decreased by GSH. This is consistent with the scheme of the TNT adduct formation with the central role of the nitroso metabolite (NO-DNT) reaction with protein or nonprotein thiols (RSH Equation 9.11) [57], The acid hydrolysis of the sulfinamide adduct (RS(0)-NH-DNT) formed after the rearrangement of the semimercaptal (RS-N(OH)-DNT Equation 9.12) will yield NH2-DNT. The mixture of NHOH-DNTs inhibits bacterial glyceraldehyde-3-phosphate dehydrogenase and glucose-6-phosphate dehydrogenase more efficiently than TNT [61]. This was attributed to the covalent modification of protein -SH groups. 

Thiol-containing enzymes are also critical targets for NO. The active-site thiol of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is subject to modification via NO-dependent reactions. This, in turn, leads to reaction with NAD, thus initiating nonenzymatic ADP-ribosylation reactions (Mohr... 

Table 12 Identification of G3PC ARATH (GLYCERALDEHYDE 3-PHOSPHATE DEHYDROGENASE, CYTOSOUC), Species Arabidopsis thaliana, SWISS-PROT (SP) entry P25858 using different PMF tools. The restricting parameters used were Arabidopsis thaliana for the species, a minimum number of 4 matched masses, a maximal tolerance for masses of 60 ppm, at most one missed cleavage of tryptic peptides allowed, and the modifications accepted were oxidised methionines and cysteines treated with iodoacetamide to form carboxyamidomethyl cysteines. The databases queried by each program are listed in the right column. In the score column, the first value is the score of the first candidate protein, followed by either the score of the second candidate protein (if the first one is the correct one)...

Mohr, S., J. S. Stamler, and B. Brune. Posttranslational modification of glyceraldehyde-3-phosphate dehydrogenase by S-nitrosylation and subsequent NADH attachment. J Biol Chem2im 1996 4209-14. 

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