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重构于平面磷脂双分子层中的人血小板表面膜上一种主要为阴离子选择性通道的传导和阻断特性。

Conduction and blocking properties of a predominantly anion-selective channel from human platelet surface membrane reconstituted into planar phospholipid bilayers.

作者信息

Manning S D, Williams A J

机构信息

Department of Cardiac Medicine, University of London, United Kingdom.

出版信息

J Membr Biol. 1989 Jul;109(2):113-22. doi: 10.1007/BF01870850.

Abstract

We have investigated the basic properties of a predominantly anion-selective channel derived from highly purified human platelet surface membrane. Single channels have been reconstituted into planar phospholipid bilayers by fusion of membrane vesicles and recorded under voltage-clamp conditions. The channel is found to have the following properties: (i) Channel activity occurs in bursts of openings separated by long closed periods. (ii) The current-voltage relationship is nonlinear. Channel current is seen to rectify, with less current flowing at positive than at negative voltages. Rectification may be due to asymmetric block by HEPES/Tris buffers. In 450 mM KCl, 5 mM HEPES/Tris, pH 7.2, the single channel conductance at -40 mV is approximately 160 pS and at +40 mV is approximately 90 pS. (iii) The conductance-concentration relationship follows a simple saturation curve. Half maximal conductance is achieved at a concentration of approximately 1000 mM KCl, and the curve saturates at a conductance of approximately 500 pS. (iv) Reversal potentials interpreted in terms of the Goldman-Hodgkin-Katz equation indicate a Cl: K permeability ratio of 4:1. (v) The channel accepts all of the halides as well as a number of other anions. The following sequence of relative anion permeabilities (in the presence of K+) is obtained: F- less than acetate- less than gluconate- less than Cl- less than Br- less than I- less than NO3- less tha SCN-.(vi) Cations as large as TEA+ are permeant. (vii) Current through the channel is blocked in the presence of DIDS, SITS and ATP, but not by Zn2+.

摘要

我们研究了一种主要为阴离子选择性通道的基本特性,该通道源自高度纯化的人血小板表面膜。通过膜囊泡融合将单通道重构到平面磷脂双分子层中,并在电压钳条件下进行记录。发现该通道具有以下特性:(i)通道活性以长时间关闭期分隔的开放脉冲形式出现。(ii)电流-电压关系是非线性的。通道电流表现出整流现象,正向电压下的电流小于负向电压下的电流。整流可能是由于HEPES/Tris缓冲液的不对称阻断所致。在450 mM KCl、5 mM HEPES/Tris、pH 7.2条件下,-40 mV时的单通道电导约为160 pS,+40 mV时约为90 pS。(iii)电导-浓度关系遵循简单的饱和曲线。大约在1000 mM KCl浓度时达到最大电导的一半,曲线在约500 pS的电导处饱和。(iv)根据戈德曼-霍奇金- Katz方程解释的反转电位表明Cl:K渗透率比为4:1。(v)该通道可接受所有卤化物以及许多其他阴离子。得到以下相对阴离子渗透率序列(在存在K+的情况下):F- < 醋酸根- < 葡萄糖酸根- < Cl- < Br- < I- < NO3- < SCN-。(vi)像TEA+这样大的阳离子是可通透的。(vii)在存在DIDS、SITS和ATP时,通过通道的电流被阻断,但不受Zn2+影响。

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