Reactivation of substrate-inactivated brain glutamate decarboxylase.
The effects of ATP and inorganic phosphate (Pi) on the reactivation of glutamate apodecarboxylase by its cofactor pyridoxal-5'-phosphate (pyridoxal-P) was studied. Apoenzyme was prepared by preincubation with glutamate. Apoenzyme prepared with glutamate alone was reactivated slowly and incompletely by adding a saturating concentration of pyridoxal-P (20 microM). Reactivation was slightly enhanced by 1-10 mM Pi. Reactivation by pyridoxal-P plus Pi was greatly enhanced by the presence of low concentrations (less than 100 microM) of ATP during the preparation of apoenzyme with glutamate. Reactivation was much lower if Pi was omitted. Enhancement of reactivation by ATP was due to its effect during apoenzyme formation, since ATP did not enhance reactivation if added only during reactivation and since the enhancing effect persisted after the removal of free ATP by chromatography on Sephadex G-25 after apoenzyme preparation and before reactivation. Reactivation was inhibited by high concentrations of ATP (greater than 100 microM), possibly by competition of ATP for the cofactor binding site. Four factors (glutamate, pyridoxal-P, ATP, and Pi) control a cycle of inactivation and reactivation that appears to be important in the regulation of brain glutamate decarboxylase.
D000255:Adenosine Triphosphate / Q000008:administration & dosage*; D000818:Animals; D001051:Apoenzymes / Q000378:metabolism*; D001059:Apoproteins / Q000378:metabolism*; D001921:Brain / Q000201:enzymology*; D002262:Carboxy-Lyases / Q000378:metabolism*; D004347:Drug Interactions; D004789:Enzyme Activation; D005968:Glutamate Decarboxylase / Q000378:metabolism*; D005971:Glutamates / Q000008:administration & dosage; D018698:Glutamic Acid; D010710:Phosphates / Q000008:administration & dosage*; D011732:Pyridoxal Phosphate / Q000008:administration & dosage*; D013552:Swine
D001051:Apoenzymes; D001059:Apoproteins; D005971:Glutamates; D010710:Phosphates; D018698:Glutamic Acid; D011732:Pyridoxal Phosphate; D000255:Adenosine Triphosphate; D002262:Carboxy-Lyases; D005968:Glutamate Decarboxylase