Actin modifies Ca2+ block of epithelial Na+ channels in planar lipid bilayers

被引:25
作者
Berdiev, BK
Latorre, R
Benos, DJ
Ismailov, II
机构
[1] Univ Alabama, Dept Physiol & Biophys, Birmingham, AL 35294 USA
[2] Univ Chile, Fac Ciencias, Dept Biol, Santiago, Chile
[3] Ctr Estudios Cient, Valdivia, Chile
关键词
D O I
10.1016/S0006-3495(01)76190-5
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The mechanism by which the cytoskeletal protein actin affects the conductance of amiloride-sensitive epithelial sodium channels (ENaC) was studied in planar lipid bilayers. In the presence of monomeric actin, we found a decrease in the single-channel conductance of alpha -ENaC that did not occur when the internal [Ca2+](free) as buffered to <10 nM. An analysis of single-channel kinetics demonstrated that Ca2+ induced the appearance of long-lived closed intervals separating bursts of channel activity, both in the presence and in the absence of actin. In the absence of actin, the duration of these bursts and the time spent by the channel in its open, but not in its short-lived closed state, were inversely proportional to [Ca2+]. This, together with a lengthening of the interburst intervals. translated into a dose-dependent decrease in the single-channel open probability. In contrast, a [Ca2+]-dependent decrease in a-ENaC conductance in the presence of actin was accompanied by lengthening of the burst intervals with no significant changes in the open or closed (both short- and long-lived) times. We conclude that Ca2+ acts as a "fast-to-intermediate" blocker when monomeric actin is present, producing a subsequent attenuation of the apparent unitary conductance of the channel.
引用
收藏
页码:2176 / 2186
页数:11
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