Cloning, characterization, and gene organization of K-Cl cotransporter from pig and human kidney and C. elegans.
Cloning, characterization, and gene organization of K-Cl cotransporter from pig and human kidney and C. elegans.
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DOI：
10.1152/ajprenal.1998.275.4.f550
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发表时间：
1998-10
期刊：
影响因子：
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通讯作者：
E. Holtzman;S Kumar;C. Faaland;F. Warner;P. Logue;S. J. Erickson;G. Ricken;J. Waldman;Shiv Kumar;P. Dunham
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文献类型：
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作者：
E. Holtzman;S Kumar;C. Faaland;F. Warner;P. Logue;S. J. Erickson;G. Ricken;J. Waldman;Shiv Kumar;P. Dunham
我们分离并鉴定了人、猪和秀丽隐杆线虫K-Cl共转运蛋白的cDNA。猪和人的同源物有94%相同，分别含有1，085和1，086个氨基酸。C. elegans K-Cl协同转运蛋白克隆（CE-KCC 1）含有1，003个氨基酸。哺乳动物K-Cl共转运蛋白与CE-KCC 1有约45%的相似性。三个克隆的亲水性分析表明典型的KCC拓扑结构模式，具有12个跨膜片段，跨膜结构域5和6之间的大细胞外环（KCC特有）和大COOH末端结构域。人KCC 1在多种组织中广泛表达。该KCC 1基因跨越23 kb，由24个外显子组成，而CE-KCC 1基因跨越3.5 kb，包含10个外显子。瞬时稳定转染的人胚肾细胞（HEK-293）表达人、猪和C. elegans K-Cl共转运蛋白满足2个（猪）或5个（人和C. elegans）的K-Cl共转运蛋白表达增加的标准。采用的标准是基础钾-氯共转运;刺激共转运肿胀，N-乙基马来酰亚胺，星形孢菌素，降低细胞镁浓度;和二次刺激钠-钾-氯共转运。
We isolated and characterized the cDNAs for the human, pig, and Caenorhabditis elegansK-Cl cotransporters. The pig and human homologs are 94% identical and contain 1,085 and 1,086 amino acids, respectively. The deduced protein of the C. elegans K-Cl cotransporter clone (CE-KCC1) contains 1,003 amino acids. The mammalian K-Cl cotransporters share ∼45% similarity with CE-KCC1. Hydropathy analyses of the three clones indicate typical KCC topology patterns with 12 transmembrane segments, large extracellular loops between transmembrane domains 5 and 6 (unique to KCC), and large COOH-terminal domains. Human KCC1 is widely expressed among various tissues. This KCC1 gene spans 23 kb and is organized in 24 exons, whereas the CE-KCC1 gene spans 3.5 kb and contains 10 exons. Transiently and stably transfected human embryonic kidney cells (HEK-293) expressing the human, pig, and C. elegans K-Cl cotransporter fulfilled two (pig) or five (human and C. elegans) criteria for increased expression of the K-Cl cotransporter. The criteria employed were basal K-Cl cotransport; stimulation of cotransport by swelling, N-ethylmaleimide, staurosporine, and reduced cell Mg concentration; and secondary stimulation of Na-K-Cl cotransport.