Title : Glutamate-dependent active-site labeling of brain glutamate decarboxylase.

Pub. Date : 1990 Aug

PMID : 1973454






6 Functional Relationships(s)
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Compound Name
Protein Name
Organism
1 Glutamate-dependent active-site labeling of brain glutamate decarboxylase. Glutamic Acid glutamate-ammonia ligase Rattus norvegicus
2 To test this prediction, the effect of glutamate on the incorporation of pyridoxal-P into rat-brain GAD was studied by incubating GAD with [32P]pyridoxal-P, followed by reduction with NaBH4 to link irreversibly the cofactor to the enzyme. Glutamic Acid glutamate-ammonia ligase Rattus norvegicus
3 Adding glutamate to the reaction mixture strongly stimulated labeling of GAD, as expected. Glutamic Acid glutamate-ammonia ligase Rattus norvegicus
4 4-Deoxypyridoxine 5"-phosphate (deoxypyridoxine-P), a close structural analogue of pyridoxal-P, was a competitive inhibitor of the activation of glutamate apodecarboxylase by pyridoxal-P (Ki = 0.27 microM) and strongly inhibited glutamate-dependent labeling of GAD. Glutamic Acid glutamate-ammonia ligase Rattus norvegicus
5 Both proteins could be purified by immunoaffinity chromatography on a column prepared with a monoclonal antibody to GAD, and both were labeled in a glutamate-dependent, deoxypyridoxine-P-sensitive manner, indicating that both were GAD. Glutamic Acid glutamate-ammonia ligase Rattus norvegicus
6 Both proteins could be purified by immunoaffinity chromatography on a column prepared with a monoclonal antibody to GAD, and both were labeled in a glutamate-dependent, deoxypyridoxine-P-sensitive manner, indicating that both were GAD. Glutamic Acid glutamate-ammonia ligase Rattus norvegicus