[Non-equivalency of active centers of alpha-ketoglutarate dehydrogenase detected by modification of histidine residues].
Using selective chemical modification of histidine residues of the alpha-ketoglutarate dehydrogenase component within the alpha-ketoglutarate dehydrogenase complex, the existence of interconvertible forms of the enzyme was demonstrated. These forms are distinguished by kinetics of inactivation caused by diethylpyrocarbonate. The interconversion of the enzyme forms involves alpha-ketoglutarate. Studies on substrate effects on the inactivation kinetics of individual enzyme forms revealed the non-equivalency of the enzyme active centers within the dimeric molecule of the alpha-ketoglutarate dehydrogenase component. The accessibility of an essential histidine residue in the active center of a neighbouring substrate-free monomer to the modifier increases as a result of interaction of the enzyme active centers during alpha-ketoglutarate binding by one of the subunits. The non-equivalency of the active centers manifests itself in different rates of interaction and in the unequal stability of binding of alpha-ketoglutarate to the alternate sites of the dimer. It is assumed that the biphasic kinetics of inactivation of pigeon breast muscle alpha-ketoglutarate dehydrogenase is due to tight binding of alpha-ketoglutarate in one of active centers of the enzyme dimeric molecule.
D000818:Animals; D001665:Binding Sites; D010856:Columbidae; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D066298:In Vitro Techniques; D007655:Ketoglutarate Dehydrogenase Complex / Q000032:analysis / Q000037:antagonists & inhibitors*; D007656:Ketoglutaric Acids / Q000494:pharmacology; D007658:Ketone Oxidoreductases / Q000037:antagonists & inhibitors*; D007700:Kinetics; D008954:Models, Biological; D009132:Muscles / Q000201:enzymology; D013379:Substrate Specificity
D007656:Ketoglutaric Acids; D007658:Ketone Oxidoreductases; D007655:Ketoglutarate Dehydrogenase Complex; D004047:Diethyl Pyrocarbonate
Neékvivalentnost' aktivnykh tsentrov alpha-ketoglutaratdegidrogenazy,vyiavliaemaia pri modifikatsii gistidinovykh ostatkov.