Grigoriev Sergey M, Muro Concepción, Dejean Laurent M, Campo Maria Luisa, Martinez-Caballero Sonia, Kinnally Kathleen W
College of Dentistry, Department of Basic Sciences, New York University, 345 East 24th Street, New York, New York 10010, USA.
Int Rev Cytol. 2004;238:227-74. doi: 10.1016/S0074-7696(04)38005-8.
Electrophysiological techniques have been integral to our understanding of protein translocation across various membranes, and, in particular, the mitochondrial inner and outer membranes. Descriptions of various methodologies (for example, patch clamp, planar bilayers, and tip dip, and their past and potential contributions) are detailed within. The activity of protein import channels of native mitochondrial inner and outer membranes can be studied by directly patch clamping mitochondria and mitoplasts (mitochondria stripped of their outer membrane by French pressing) from various genetically manipulated strains of yeast and mammalian tissue cultured cells. The channel activities of TOM, TIM23, and TIM22 complexes are compared with those reconstituted in proteoliposomes and with those of the recombinant proteins Tom40p, Tim23p, and Tim22p, which play major roles in protein translocation. Studies of the mechanism(s) and the role of channels in protein translocation in mitochondria are prototypes, as the same principles are likely followed in all biological membranes including the endoplasmic reticulum and chloroplasts. The ability to apply electrophysiological techniques to these channels is now allowing investigations into the role of mitochondria in diverse fields such as neurotransmitter release, long-term potentiation, and apoptosis.
电生理技术对于我们理解蛋白质跨各种膜的转运，尤其是线粒体内外膜的转运，不可或缺。本文详细描述了各种方法（例如，膜片钳、平面双层膜和尖端浸入法，以及它们过去和潜在的贡献）。通过直接膜片钳记录来自各种基因操作的酵母菌株和哺乳动物组织培养细胞的线粒体和线粒体球（通过法式压榨去除外膜的线粒体），可以研究天然线粒体内外膜蛋白质导入通道的活性。将TOM、TIM23和TIM22复合物的通道活性与在蛋白脂质体中重组的通道活性以及在蛋白质转运中起主要作用的重组蛋白Tom40p、Tim23p和Tim22p的通道活性进行比较。线粒体中蛋白质转运通道的机制和作用的研究是典型的，因为包括内质网和叶绿体在内的所有生物膜可能都遵循相同的原理。现在，将电生理技术应用于这些通道的能力使得人们能够研究线粒体在神经递质释放、长时程增强和细胞凋亡等不同领域中的作用。
