BRAIN GLUTAMATE-DECARBOXYLASE CLONED IN LAMBDA-GT-11 - FUSION PROTEIN PRODUCES GAMMA-AMINOBUTYRIC-ACID
BRAIN GLUTAMATE-DECARBOXYLASE CLONED IN LAMBDA-GT-11 - FUSION PROTEIN PRODUCES GAMMA-AMINOBUTYRIC-ACID
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DOI：
10.1126/science.3518061
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发表时间：
1986-05-30
期刊：
影响因子：
56.9
通讯作者：
TOBIN, AJ
中科院分区：
文献类型：
--
作者：
KAUFMAN, DL;MCGINNIS, JF;TOBIN, AJ
谷氨酸脱羧酶（GAD; E.C. 4.1.1.15）将谷氨酸盐转化为γ。氨基丁酸（GABA）是脊椎动物中枢神经系统中的主要抑制性神经递质。本报告描述了通过免疫学筛选λ gt-11脑互补DNA表达文库来分离GAD互补DNA克隆。该克隆产生的融合蛋白催化谷氨酸转化为GABA和二氧化碳，证实其身份为GAD。针对β-半乳糖苷酶从溶液中去除GAD酶活性，表明该活性与融合蛋白相关。在免疫印迹实验中，所有三种可用的GAD抗血清与融合多肽和脑提取物中的两种主要多肽（分子大小，60，000和66，000道尔顿）反应。
Glutamate decarboxylase (GAD; E.C. 4.1.1.15) 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.