Physicochemical evidence for the existence of two pyridoxal 5'-phosphate binding sites on glutamate dehydrogenase and characterization of their functional role.
Kinetic studies of pyridoxal 5'-phosphate binding to glutamate dehydrogenase (EC 1.4.1.3) has provided evidence for two specific binding sites, chemically identified as Lys 126 and Lys 333. Use of protecting ligands permitted the selective modification of only one of these lysines, and showed that (1) Lys 333 modification results in depolymerisation of the enzyme into active hexamers; (2) Lys 126-modified enzyme was 92% inactivated. The residual activity was desensitized to GTP. The inactivation process was cooperative, maximum inactivation occurring as soon as half of the Lys 126 were modified.
D000818:Animals; D001665:Binding Sites; D002417:Cattle; D005969:Glutamate Dehydrogenase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007656:Ketoglutaric Acids / Q000494:pharmacology; D007700:Kinetics; D008024:Ligands; D008099:Liver / Q000201:enzymology; D008239:Lysine / Q000378:metabolism; D008970:Molecular Weight; D009249:NADP / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D011732:Pyridoxal Phosphate / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000494:pharmacology
D007656:Ketoglutaric Acids; D008024:Ligands; D009249:NADP; D011732:Pyridoxal Phosphate; D006160:Guanosine Triphosphate; D005969:Glutamate Dehydrogenase; D008239:Lysine