Schein S J, Colombini M, Finkelstein A
J Membr Biol. 1976 Dec 28;30(2):99-120. doi: 10.1007/BF01869662.
We have incorporated into planar lipid bilayer membranes a voltage-dependent, anion-selective channel (VDAC) obtained from Paramecium aurelia. VDAC-containing membranes have the following properties: (1) The steady-state conductance of a many-channel membrane is maximal when the transmembrane potential is zero and decreases as a steep function of both positive and negative voltage. (2) The fraction of time that an individual channel stays open is strongly voltage dependent in a manner that parallels the voltage dependence of a many-channel membrane. (3) The conductance of the open channel is about 500 pmho in 0.1 to 1.0 M salt solutions and is ohmic. (4) The channel is about 7 times more permeable to Cl- than to K+ and is impermeable to Ca++. The procedure for obtaining VDAC; AND THE PROPERTIES OF THE CHANNEL ARE HIGHLY REPRODUCIBLE. VDAC activity was found, upon fractionation of the paramecium membranes, to come from the mitochondria. We note that the published data on mitochondrial Cl- permeability suggest that there may indeed be a voltage-dependent Cl- permeability in mitochondria. The method of incorporating VDAC into planar lipid bilayers may be generally useful for reconstituting biological transport systems in these membranes.
我们已将从双小核草履虫获得的电压依赖性阴离子选择性通道(VDAC)整合到平面脂质双分子层膜中。含有VDAC的膜具有以下特性:(1)当跨膜电位为零时,多通道膜的稳态电导最大,并随着正电压和负电压的急剧变化而降低。(2)单个通道保持开放的时间比例强烈依赖于电压,其方式与多通道膜的电压依赖性相似。(3)在0.1至1.0 M盐溶液中,开放通道的电导约为500皮西门子,且呈欧姆特性。(4)该通道对Cl-的通透性比对K+高约7倍,对Ca++不通透。获得VDAC的方法;并且通道的特性具有高度可重复性。在对草履虫膜进行分级分离时发现,VDAC活性来自线粒体。我们注意到,已发表的关于线粒体Cl-通透性的数据表明,线粒体中确实可能存在电压依赖性Cl-通透性。将VDAC整合到平面脂质双分子层中的方法可能普遍适用于在这些膜中重建生物转运系统。