CLONING, EXPRESSION, AND LOCALIZATION OF A RAT-BRAIN HIGH-AFFINITY GLYCINE TRANSPORTER
CLONING, EXPRESSION, AND LOCALIZATION OF A RAT-BRAIN HIGH-AFFINITY GLYCINE TRANSPORTER
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DOI：
10.1073/pnas.89.15.7189
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发表时间：
1992-08-01
影响因子：
11.1
通讯作者：
DAVIDSON, N
中科院分区：
文献类型：
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作者：
GUASTELLA, J;BRECHA, N;DAVIDSON, N
采用PCR和噬斑杂交相结合的方法，从大鼠脑中克隆了一个编码甘氨酸转运蛋白的cDNA。从这个克隆（指定GLYT 1）合成的mRNA指导表达的钠和氯依赖性，高亲和力摄取[H-3]甘氨酸爪蟾卵母细胞。[H-3]由克隆GLYT 1介导的甘氨酸转运被肌氨酸阻断，但不被甲氨基异丁酸或L-丙氨酸阻断，底物特异性类似于先前鉴定的称为系统Gly的甘氨酸摄取系统。原位杂交显示，GLYT 1显着表达在颈脊髓和脑干，两个区域的中枢神经系统，其中甘氨酸是一个假定的神经递质。GLYT 1也在小脑和嗅球中强烈表达，在其他大脑区域中表达水平较低。GLYT 1 cDNA的开放阅读框预测了一种含有633个氨基酸的蛋白质，其分子量几乎等于70 kDa。GLYT 1蛋白的一级结构和亲水性特征表明，该蛋白是钠和氯依赖性转运蛋白超家族的成员，其利用神经递质和相关物质作为底物。
A cDNA clone encoding a glycine transporter has been isolated from rat brain by a combined PCR and plaque-hybridization strategy. mRNA synthesized from this clone (designated GLYT1) directs the expression of sodium- and chloride-dependent, high-affinity uptake of [H-3]glycine by Xenopus oocytes. [H-3]Glycine transport mediated by clone GLYT1 is blocked by sarcosine but is not blocked by methylaminoisobutyric acid or L-alanine, a substrate specificity similar to that described for a previously identified glycine-uptake system called system Gly. In situ hybridization reveals that GLYT1 is prominently expressed in the cervical spinal cord and brainstem, two regions of the central nervous system where glycine is a putative neurotransmitter. GLYT1 is also strongly expressed in the cerebellum and olfactory bulb and is expressed at lower levels in other brain regions. The open reading frame of the GLYT1 cDNA predicts a protein containing 633 amino acids with a molecular mass of almost-equal-to 70 kDa. The primary structure and hydropathicity profile of GLYT1 protein reveal that this protein is a member of the sodium- and chloride-dependent superfamily of transporters that utilize neurotransmitters and related substances as substrates.