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一种来自非洲爪蟾卵母细胞膜的钙离子激活通道被重组到平面双层膜中。

A Ca2+-activated channel from Xenopus laevis oocyte membranes reconstituted into planar bilayers.

作者信息

Young G P, Young J D, Deshpande A K, Goldstein M, Koide S S, Cohn Z A

出版信息

Proc Natl Acad Sci U S A. 1984 Aug;81(16):5155-9. doi: 10.1073/pnas.81.16.5155.

Abstract

Plasma membrane fractions from Xenopus laevis oocytes were incorporated into planar lipid bilayers. We show the existence of numerous Ca2+-activated nonspecific channels that are more permeable to anions. These channels are activated by Ca2+ at micromolar concentration but not by Mg2+, Zn2+, or Mn2+, even at millimolar concentrations. Decreasing Ca2+ concentration to less than 1 microM decreases the time of channel opening until channels close completely in the absence of Ca2+ and in the presence of EGTA. I- and Br- are more permeable through this channel than Cl-. The time during which the channels remain open is also voltage-dependent, with the channels switching off at higher voltages in both polarities. Single-channel activity shows a conductance of 380 pS in 1 M NaCl and 1 mM CaCl2, with an average open lifetime of 1.5 s at 40 mV. Similar channels are found in different stages of oocyte maturation. These observations support the hypothesis that an increase in oocyte-free Ca2+ activates directly these channels, and the resultant Cl- efflux forms the ionic basis for the fertilization potential in X. laevis.

摘要

非洲爪蟾卵母细胞的质膜组分被整合到平面脂质双分子层中。我们发现存在大量Ca2+激活的非特异性通道,这些通道对阴离子的通透性更高。这些通道在微摩尔浓度的Ca2+作用下被激活,但即使在毫摩尔浓度下,Mg2+、Zn2+或Mn2+也不能激活它们。将Ca2+浓度降低到小于1微摩尔会减少通道开放时间,直到在没有Ca2+和存在乙二醇双(2-氨基乙基醚)四乙酸(EGTA)的情况下通道完全关闭。碘离子(I-)和溴离子(Br-)通过该通道的通透性比氯离子(Cl-)更高。通道保持开放的时间也依赖于电压,在两种极性下,通道在较高电压时关闭。单通道活性在1 M氯化钠(NaCl)和1 mM氯化钙(CaCl2)中显示出380皮安(pS)的电导,在40毫伏(mV)时平均开放寿命为1.5秒。在卵母细胞成熟的不同阶段发现了类似的通道。这些观察结果支持这样的假说,即卵母细胞中游离Ca2+的增加直接激活这些通道,并且由此产生的Cl-外流形成了非洲爪蟾受精电位的离子基础。

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