Effects of aldosterone on biosynthesis, traffic, and functional expression of epithelial sodium channels in A6 cells.

Effects of aldosterone on biosynthesis, traffic, and functional expression of epithelial sodium channels in A6 cells.
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DOI:
10.1085/jgp.20028559
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发表时间:
2002-05
期刊:
The Journal of general physiology
影响因子:
--
通讯作者:
Canessa CM
Canessa CM
中科院分区:
其他
文献类型:
--
作者:
Alvarez de la Rosa D;Li H;Canessa CM

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集合管通过调节上皮 Na+ 通道 (ENaC) 的丰度/活性来调节 Na+ 转运。在这项研究中,我们利用 A6 细胞系中表达的内源通道研究了 ENaC 的合成、降解、内吞作用和活性,以及​​醛固酮对这些过程的影响。使用一组新产生的针对两栖动物 ENaC 三个亚基的特异性抗体进行了生化研究。我们的结果表明,醛固酮可同时转录和翻译 α、β 和 γ 亚基,并增强这两个过程:分别增加两倍和四倍。新通道的生物合成之后,可以在 α 和 β 亚基中获得内切糖苷酶 H 抗性寡糖,并且在 α 的情况下,出现对还原剂具有抗性的形式。整个亚基池的半衰期(t 1/2 40-70 分钟)比顶膜通道部分(t 1/2 12-17 分钟)长。醛固酮诱导顶膜中三个亚基的丰度增加四倍,而开放概率、单通道动力学或 ENaC 亚基的降解率没有显着变化。因此,醛固酮反应可以通过顶端通道丰度的增加来解释,这至少部分归因于亚基的从头合成。
The collecting duct regulates Na+ transport by adjusting the abundance/activity of epithelial Na+ channels (ENaC). In this study we have investigated the synthesis, degradation, endocytosis, and activity of ENaC and the effects of aldosterone on these processes using endogenous channels expressed in the A6 cell line. Biochemical studies were performed with a newly raised set of specific antibodies against each of the three subunits of the amphibian ENaC. Our results indicate simultaneous transcription and translation of α, β, and γ subunits and enhancement of both processes by aldosterone: two- and fourfold increase, respectively. The biosynthesis of new channels can be followed by acquisition of endoglycosidase H–resistant oligosacharides in α and β subunits and, in the case of α, by the appearance of a form resistant to reducing agents. The half-life of the total pool of subunits (t 1/2 40–70 min) is longer than the fraction of channels in the apical membrane (t 1/2 12–17 min). Aldosterone induces a fourfold increase in the abundance of the three subunits in the apical membrane without significant changes in the open probability, kinetics of single channels, or in the rate of degradation of ENaC subunits. Accordingly, the aldosterone response could be accounted by an increase in the abundance of apical channels due, at least in part, to de novo synthesis of subunits.
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