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将美西螈肠上皮细胞基底外侧膜中的内向整流钾通道重组到平面脂质双分子层中。

Reconstitution of an inwardly rectifying potassium channel from the basolateral membranes of Necturus enterocytes into planar lipid bilayers.

作者信息

Costantin J, Alcalen S, de Souza Otero A, Dubinsky W P, Schultz S G

机构信息

Department of Physiology and Cell Biology, University of Texas Medical School, Houston 77225.

出版信息

Proc Natl Acad Sci U S A. 1989 Jul;86(13):5212-6. doi: 10.1073/pnas.86.13.5212.

Abstract

Basolateral membrane vesicles from Necturus enterocytes, highly (greater than 20-fold) enriched in Na+,K+-ATPase, were reconstituted into planar lipid bilayers. The principal channel activity observed is selective for K+ over Na+ and Cl-. This K+ channel is blocked by Ba2+ and Leiurus quinquestriatus venom but is not affected by Ca2+ over the range of 10(-3) to less than 10(-7) M and is not inhibited by charybdotoxin. L. quinquestriatus venom also markedly reduces the conductance of the basolateral membrane of intact villus cells of Necturus small intestine. The open-time probability (Po) of the channel displays a voltage-dependence characteristic of an "inward rectifier"; i.e., the channel inactivates when the basolateral membrane is depolarized and Po increases with increasing hyperpolarization of that barrier. Assuming that similar properties prevail under physiological conditions, this characteristic could provide, in part, an explanation for the parallelism between Na+-pump and K+-leak activities of the basolateral membrane observed in this epithelium. Thus, an increase in rheogenic Na+-pump activity at the basolateral membrane would hyperpolarize that barrier and, in turn, increase the open time of this K+ channel.

摘要

将富含Na⁺,K⁺-ATP酶(富集程度超过20倍)的美西螈肠上皮细胞基底外侧膜囊泡重构成平面脂双层。观察到的主要通道活性对K⁺的选择性高于Na⁺和Cl⁻。该K⁺通道被Ba²⁺和金环蛇毒阻断,但在10⁻³至小于10⁻⁷M的Ca²⁺浓度范围内不受影响,也不被蝎毒素抑制。金环蛇毒还显著降低了美西螈小肠完整绒毛细胞基底外侧膜的电导。该通道的开放时间概率(Po)表现出“内向整流器”的电压依赖性特征;即,当基底外侧膜去极化时通道失活,且Po随着该屏障超极化程度的增加而增加。假设在生理条件下也具有类似特性,这一特征可以部分解释在该上皮细胞中观察到的基底外侧膜Na⁺泵和K⁺泄漏活性之间的平行关系。因此,基底外侧膜上生电Na⁺泵活性的增加会使该屏障超极化,进而增加该K⁺通道的开放时间。

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