Thomas L, Blachly-Dyson E, Colombini M, Forte M
Department of Zoology, University of Maryland, College Park 20742.
Proc Natl Acad Sci U S A. 1993 Jun 15;90(12):5446-9. doi: 10.1073/pnas.90.12.5446.
Voltage-gated ion-channel proteins contain "voltage-sensing" domains that drive the conformational transitions between open and closed states in response to changes in transmembrane voltage. We have used site-directed mutagenesis to identify residues affecting the voltage sensitivity of a mitochondrial channel, the voltage-dependent anion-selective channel (VDAC). Although charge changes at many sites had no effect, at other sites substitutions that increased positive charge also increased the steepness of voltage dependence and substitutions that decreased positive charge decreased voltage dependence by an appropriate amount. In contrast to the plasma membrane K+ and Na+ channels, these residues are distributed over large parts of the VDAC protein. These results have been used to define the conformational transitions that accompany voltage gating of an ion channel. This gating mechanism requires the movement of large portions of the VDAC protein through the membrane.
电压门控离子通道蛋白包含“电压感应”结构域,该结构域可响应跨膜电压的变化驱动通道在开放态和关闭态之间发生构象转变。我们利用定点诱变来鉴定影响线粒体通道(电压依赖性阴离子选择性通道,VDAC)电压敏感性的残基。尽管许多位点的电荷变化没有影响,但在其他位点,增加正电荷的取代也增加了电压依赖性的陡度,而减少正电荷的取代则使电压依赖性相应降低。与质膜钾离子和钠离子通道不同,这些残基分布在VDAC蛋白的大部分区域。这些结果已被用于定义离子通道电压门控所伴随的构象转变。这种门控机制需要VDAC蛋白的大部分区域穿过膜。