Isolation and characterization of a Na-H antiporter-deficient mutant of LLC-PK1 cells.
Isolation and characterization of a Na-H antiporter-deficient mutant of LLC-PK1 cells.
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LLC-PK1 细胞 Na-H 逆向转运蛋白缺陷突变体的分离和表征。
DOI：
10.1152/ajpcell.1986.251.5.c825
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Slayman,CW
中科院分区：
文献类型：
--
作者：
Agarwal,N;Haggerty,JG;Adelberg,EA;Slayman,CW
LLC-PK 1是一种来源于猪肾上皮的已建立的细胞系;它在体外分化形成极化的上皮片层，能够介导溶质和水的转运。我们已经使用了Pouyssegur等人（Proc.Natl. Acad. Sci. USA. 81：4833-4837，1984）以分离Na-H反向转运蛋白活性缺陷的LLC-PK 1细胞的突变体。该突变体在pH 7.0及以上的存在和不存在碳酸氢盐的情况下正常生长;然而，在pH 6.5时，它需要碳酸氢盐来生长。因此，Na-H反向转运蛋白的细胞质碱化对于在不存在碳酸氢盐的酸性pH下的生长是必不可少的。在不存在碳酸氢盐的情况下，反向转运蛋白对于圆顶的形成（矢量水运输的结果）也是必不可少的。
LLC-PK1 is an established cell line derived from pig kidney epithelium; it differentiates in vitro to form a polarized epithelial sheet, capable of the vectorial transport of solutes and water. We have used a modification of the "proton-suicide" method of Pouyssegur et al. (Proc. Natl. Acad. Sci. USA. 81:4833-4837, 1984) to isolate a mutant of LLC-PK1 cells that is deficient in Na-H antiporter activity. The mutant grows normally at pH 7.0 and above in the presence and absence of bicarbonate; at pH 6.5, however, it requires bicarbonate for growth. Alkalinization of the cytoplasm by the Na-H antiporter thus appears to be essential for growth at acidic pH in the absence of bicarbonate. The antiporter is also essential for the formation of domes (a consequence of vectorial water transport) in the absence of bicarbonate.