Specific modification of the GTP binding sites of rat 5'-adenylic acid aminohydrolase by periodate-oxidized GTP.
1. Rat skeletal muscle AMP deaminase (AMP aminohydrolase, EC3.5.4.6) can be inactivated by incubation with the periodate-oxidized analogue of the enzyme inhibitor GTP. 2. Nucleoside triphosphates and KCl at high concentrations protect against inactivation, while ADP has no effect. 3. The inactivation can be reversed by the addition of GTP and amino acids and made irreversible by reduction with NaBH4. This indicates that, in the binding of the oxidized GTP to the enzyme, a Schiff base is formed between the aldehyde groups of the inhibitor and amino groups of the enzyme. 4. The kinetic properties of the reduced (oxidized GTP)-AMP deaminase derivative indicate that the loss of activity results from an increase in Km while no appreciable change in V is observed; consequently, the enzyme shows positive homotropic cooperativity even in the presence of optimal KCl concentration. 5. Since the treated enzyme shows kinetic properties similar to those of the native enzyme in the presence of GTP, and since the loss of sensitivity to GTP is directly proportional to the degree of inactivation, it is concluded that the oxidized GTP specifically modifies the binding sites for GTP. 6. Binding of the radioactive oxidized GTP shows that two binding sites for this reagent exist in the AMP deaminase molecule.
D000659:AMP Deaminase / Q000037:antagonists & inhibitors*; D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D001894:Borohydrides; D006160:Guanosine Triphosphate / Q000494:pharmacology*; D007700:Kinetics; D009132:Muscles / Q000201:enzymology; D009709:Nucleotide Deaminases / Q000037:antagonists & inhibitors*; D010084:Oxidation-Reduction; D010504:Periodic Acid; D011189:Potassium Chloride / Q000494:pharmacology; D011485:Protein Binding; D051381:Rats; D014633:Valine / Q000494:pharmacology
D001894:Borohydrides; D010504:Periodic Acid; D000244:Adenosine Diphosphate; D011189:Potassium Chloride; D006160:Guanosine Triphosphate; D009709:Nucleotide Deaminases; D000659:AMP Deaminase; D014633:Valine