Purification of L-glutamate decarboxylase by affinity chromatography.
L-Glutamate decarboxylase (L-glutamate 1-carboxy-lyase, EC 4.1.1.15) from rat brain synaptosomal extract was partially purified by affinity chromatography. On further purification by DEAE-Sephadex A 50 and Sephadex G-200, L-glutamate decarboxylase was purified to greater extent. It was found that a single affinity chromatography by appropriate elution gave a highly purified protein giving a single band of high specific activity on polyacrylamide gradient gel slab electrophoresis with minimal contamination. Substrate specificity of the purified enzyme was modified in the presence of 6-azauracil or phenylalanine resulting in decreased specificity to L-glutamate and increased specificity to L-aspartate.
D000818:Animals; D001224:Aspartic Acid; D001921:Brain / Q000201:enzymology*; D002262:Carboxy-Lyases / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D003609:Dactinomycin / Q000494:pharmacology; D005968:Glutamate Decarboxylase / Q000302:isolation & purification* / Q000378:metabolism; D009474:Neurons / Q000201:enzymology; D010649:Phenylalanine / Q000494:pharmacology; D051381:Rats; D014498:Uracil / Q000031:analogs & derivatives / Q000494:pharmacology
D003609:Dactinomycin; D001224:Aspartic Acid; D010649:Phenylalanine; D014498:Uracil; D002262:Carboxy-Lyases; D005968:Glutamate Decarboxylase