Brain glutamate decarboxylase cloned in lambda gt-11: fusion protein produces gamma-aminobutyric acid

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Science  30 May 1986:
Vol. 232, Issue 4754, pp. 1138-1140
DOI: 10.1126/science.3518061


Glutamate decarboxylase (GAD; E.C. converts glutamate to gamma-aminobutyric acid (GABA), the major inhibitory neurotransmitter in the vertebrate central nervous system. This report describes the isolation of a GAD complementary DNA clone by immunological screening of a lambda gt-11 brain complementary DNA expression library. The fusion protein produced by this clone catalyzes the conversion of glutamate to GABA and carbon dioxide, confirming its identity as GAD. Antibodies to beta-galactosidase remove GAD enzymatic activity from solution, showing that this activity is associated with the fusion protein. In immunoblotting experiments all three available antisera to GAD reacted with the fusion polypeptide and with two major polypeptides (molecular size, 60,000 and 66,000 daltons) in brain extracts.

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