Lee W S, Wells R G, Sabbag R V, Mohandas T K, Hediger M A
Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115.
J Clin Invest. 1993 May;91(5):1959-63. doi: 10.1172/JCI116415.
We have recently cloned, sequenced, and characterized a rat kidney cDNA (D2) that stimulates cystine as well as dibasic and neutral amino acid transport. In order to evaluate the role of this protein in human inherited diseases such as cystinuria, we have isolated a human D2 clone (D2H) by low stringency screening of a human kidney cDNA library using the radiolabeled D2 insert as a probe. The D2H cDNA is 2284 nucleotides long and encodes a 663 amino acid protein that is 80% identical to the rat D2 amino acid sequence and 86% to that of the rabbit homologue rBAT. Microinjection of in vitro transcribed D2H cRNA into Xenopus oocytes induced uptake of cystine as well as dibasic and neutral amino acids in a pattern similar to that of rat D2 and rabbit rBAT. Both neutral and dibasic amino acids inhibited the D2H-induced uptake of cystine. Northern blot analysis demonstrated that D2H, like D2 and rBAT, is expressed strongly in the kidney and intestine. Southern blot analysis of genomic DNA from a panel of mouse-human somatic cell hybrids showed that the human gene for D2H resides on chromosome 2.
我们最近克隆、测序并鉴定了一种大鼠肾脏cDNA（D2），它能刺激胱氨酸以及二碱基和中性氨基酸的转运。为了评估这种蛋白质在诸如胱氨酸尿症等人类遗传性疾病中的作用，我们使用放射性标记的D2插入片段作为探针，通过对人肾脏cDNA文库进行低严谨度筛选，分离出了一个人类D2克隆（D2H）。D2H cDNA长2284个核苷酸，编码一种663个氨基酸的蛋白质，该蛋白质与大鼠D2氨基酸序列的同源性为80%，与兔同源物rBAT的同源性为86%。将体外转录的D2H cRNA显微注射到非洲爪蟾卵母细胞中，诱导胱氨酸以及二碱基和中性氨基酸的摄取，其模式与大鼠D2和兔rBAT相似。中性和二碱基氨基酸均抑制D2H诱导的胱氨酸摄取。Northern印迹分析表明，D2H与D2和rBAT一样，在肾脏和肠道中强烈表达。对一组小鼠 - 人类体细胞杂种的基因组DNA进行Southern印迹分析表明，人类D2H基因位于2号染色体上。
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