鉴定钠钾ATP酶α亚基中一个与哇巴因敏感性差异有关的区域。 - Suppr | 超能文献

鉴定钠钾ATP酶α亚基中一个与哇巴因敏感性差异有关的区域。

Identification of a region within the Na,K-ATPase alpha subunit that contributes to differential ouabain sensitivity.

作者信息

Emanuel J R, Graw S, Housman D, Levenson R

机构信息

Department of Cell Biology, Yale University School of Medicine, New Haven, Connecticut 06510.

出版信息

Mol Cell Biol. 1989 Sep;9(9):3744-9. doi: 10.1128/mcb.9.9.3744-3749.1989.

DOI:10.1128/mcb.9.9.3744-3749.1989

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC362435/

Abstract

To analyze determinants within the Na,K-ATPase alpha subunit that contribute to differential ouabain sensitivity, we constructed and expressed a panel of chimeric cDNA molecules between ouabain-resistant and ouabain-sensitive alpha subunit cDNAs. When introduced into ouabain-sensitive monkey CV-1 cells, ouabain-resistant rat alpha 1 subunit cDNA and chimeras in which the 5' end of ouabain-sensitive human alpha 1 or rat alpha 2 subunit cDNA was replaced by the 5' end of rat alpha 1 subunit cDNA conferred resistance to 100 microM ouabain. Monkey cells transfected with the reciprocal chimeras were unable to survive selection in 1 microM ouabain. Rat alpha 2 subunit cDNA and a chimera in which the 5' end of rat alpha 1 subunit cDNA was replaced by the 5' end of rat alpha 2 subunit cDNA conferred resistance to 0.5 microM ouabain. These results suggest that determinants of ouabain resistance reside within the amino-terminal portions of the rat alpha 1 and alpha 2 subunits. Expression of chimeric alpha subunit cDNAs should prove useful for elucidating the structural basis of Na,K-ATPase function.

摘要

为了分析钠钾ATP酶α亚基中导致哇巴因敏感性差异的决定因素，我们构建并表达了一组在耐哇巴因和敏感的α亚基cDNA之间的嵌合cDNA分子。当将耐哇巴因的大鼠α1亚基cDNA以及人敏感的α1或大鼠敏感的α2亚基cDNA的5′端被大鼠α1亚基cDNA的5′端取代的嵌合体导入敏感的猴CV-1细胞时，细胞对100微摩尔哇巴因产生抗性。用反向嵌合体转染的猴细胞在1微摩尔哇巴因中无法在选择中存活。大鼠α2亚基cDNA以及大鼠α1亚基cDNA的5′端被大鼠α2亚基cDNA的5′端取代的嵌合体对0.5微摩尔哇巴因产生抗性。这些结果表明，耐哇巴因的决定因素存在于大鼠α1和α2亚基的氨基末端部分。嵌合α亚基cDNA的表达对于阐明钠钾ATP酶功能的结构基础应是有用的。