通过酵母转运突变体的互补作用对哺乳动物氨基酸转运体或调节剂进行表达克隆。
Expression cloning of a mammalian amino acid transporter or modifier by complementation of a yeast transport mutant.
作者信息
Lin G, Cai X, Johnstone R M
机构信息
Department of Biochemistry, McGill University, Montreal, Quebec, Canada.
出版信息
J Cell Physiol. 1997 Dec;173(3):351-60. doi: 10.1002/(SICI)1097-4652(199712)173:3<351::AID-JCP7>3.0.CO;2-M.
A cDNA clone named L21 was isolated from L6 rat muscle cells by complementation of a yeast proline transport mutant. L21 cDNA has 2,268 bp and codes for a peptide of 228 amino acids with four potential transmembrane domains. The amino acid sequence of L21 shows no homology to any known proteins. Expression of L21 cDNA enables the mutant yeast to grow in proline as the sole nitrogen source and to transport proline, alpha-aminoisobutyric acid (AIB) and leucine. The Km for proline is about 1.0 mM. The substrate specificity of L21 expressed in yeast shows no striking similarity to known mammalian amino acid transporters.
通过对酵母脯氨酸转运突变体进行互补，从L6大鼠肌肉细胞中分离出一个名为L21的cDNA克隆。L21 cDNA有2268个碱基对，编码一个含有四个潜在跨膜结构域的228个氨基酸的肽段。L21的氨基酸序列与任何已知蛋白质均无同源性。L21 cDNA的表达使突变酵母能够以脯氨酸作为唯一氮源生长，并转运脯氨酸、α-氨基异丁酸（AIB）和亮氨酸。脯氨酸的米氏常数约为1.0 mM。在酵母中表达的L21的底物特异性与已知的哺乳动物氨基酸转运体没有明显相似性。
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