Inactivation of glutamate dehydrogenase and glutamate synthase from Bacillus megaterium by phenylglyoxal, butane-2,3-dione and pyridoxal 5'-phosphate.
Reaction of phenylglyoxal with glutamate dehydrogenase (EC 1.4.1.4), but not with glutamate synthase (EC 2.6.1.53), from Bacillus megaterium resulted in complete loss of enzyme activity. NADPH alone or together with 2-oxoglutarate provided substantial protection from inactivation by phenylglyoxal. Some 2mol of [14C]Phenylglyoxal was incorporated/mol of subunit of glutamate dehydrogenase. Addition of 1mM-NADPH decreased incorporation by 0.7mol. The Ki for phenylglyoxal was 6.7mM and Ks for competition with NADPH was 0.5mM. Complete inactivation of glutamate dehydrogenase by butane-2,3-dione was estimated by extrapolation to result from the loss of 3 of the 19 arginine residues/subunit. NADPH, but not NADH, provided almost complete protection against inactivation. Butane-2,3-dione had only a slight inactivating effect on glutamate synthase. The data suggest that an essential arginine residue may be involved in the binding of NADPH to glutamate dehydrogenase. The enzymes were inactivated by pyridoxal 5'-phosphate and this inactivation increased 3--4-fold in the borate buffer. NADPH completely prevented inactivation by pyridoxal 5'-phosphate.
D000447:Aldehydes / Q000494:pharmacology*; D001120:Arginine; D001410:Bacillus megaterium / Q000201:enzymology*; D001665:Binding Sites; D002074:Butanones / Q000494:pharmacology*; D003931:Diacetyl / Q000494:pharmacology*; D005969:Glutamate Dehydrogenase / Q000037:antagonists & inhibitors*; D005970:Glutamate Synthase / Q000037:antagonists & inhibitors*; D006037:Glyoxal / Q000031:analogs & derivatives / Q000494:pharmacology*; D007700:Kinetics; D009249:NADP / Q000378:metabolism; D011732:Pyridoxal Phosphate / Q000494:pharmacology*; D000637:Transaminases / Q000037:antagonists & inhibitors*
D000447:Aldehydes; D002074:Butanones; D006037:Glyoxal; D009249:NADP; D011732:Pyridoxal Phosphate; D001120:Arginine; D005970:Glutamate Synthase; D005969:Glutamate Dehydrogenase; D000637:Transaminases; D003931:Diacetyl