Kobayashi Y, Kaufman D L, Tobin A J
J Neurosci. 1987 Sep;7(9):2768-72. doi: 10.1523/JNEUROSCI.07-09-02768.1987.
Glutamic acid decarboxylase (GAD;E.C. 4.1.1.15) catalyzes the production of GABA, the major inhibitory neurotransmitter in the mammalian brain. We recently isolated a lambda gt-11 recombinant, lambda-GAD, that contains the cDNA for GAD from feline brain (Kaufman et al., 1986). Interestingly, the beta-galactosidase-GAD fusion protein encoded by lambda GAD is enzymatically active, catalyzing the conversion of glutamate to CO2 and GABA. Here we report the nucleotide sequence of feline GAD cDNA. It consists of 2265 bases, with a continuous open reading frame of 625 codons. The derived sequence contains the sequence Asn-Pro-His-Lys, which is identical to sequence at the pyridoxal phosphate-binding site of porcine DOPA decarboxylase (Bossa et al., 1977). The first ATG sequence in the open reading frame begins at nucleotide residue 118. The 585 codons 3' to this putative initiation site predict an amino acid composition, N-terminal residue, and molecular size consistent with published characterizations of GAD.
谷氨酸脱羧酶(GAD;E.C. 4.1.1.15)催化生成γ-氨基丁酸(GABA),它是哺乳动物大脑中的主要抑制性神经递质。我们最近分离出一种λgt-11重组体,λ-GAD,它包含来自猫脑的GAD cDNA(考夫曼等人,1986年)。有趣的是,由λ-GAD编码的β-半乳糖苷酶-GAD融合蛋白具有酶活性,催化谷氨酸转化为二氧化碳和GABA。在此我们报告猫GAD cDNA的核苷酸序列。它由2265个碱基组成,有一个625个密码子的连续开放阅读框。推导的序列包含天冬酰胺-脯氨酸-组氨酸-赖氨酸序列,这与猪多巴脱羧酶的磷酸吡哆醛结合位点的序列相同(博萨等人,1977年)。开放阅读框中的第一个ATG序列始于核苷酸残基118。该假定起始位点下游的585个密码子预测的氨基酸组成、N端残基和分子大小与已发表的GAD特征一致。