Berger Thomas K, Isacoff Ehud Y
Research Center Caesar, Ludwig-Erhard-Allee 2, 53175, Bonn, Germany,
Methods Mol Biol. 2015;1266:93-106. doi: 10.1007/978-1-4939-2272-7_6.
Understanding the function of ion channels is a major goal of molecular neurophysiology. While standard electrophysiological methods are invaluable tools to investigate the gating of ion channels, the structural rearrangements that mediate the way a channel senses physiological signals and opens and closes its gates cannot be measured electrically in a direct way. Here, we describe a method, based on site-specific labeling of a channel of interest with an environmentally sensitive fluorophore, which makes it possible to monitor conformational changes of ion channels in biological membranes in real time.
了解离子通道的功能是分子神经生理学的一个主要目标。虽然标准的电生理方法是研究离子通道门控的宝贵工具,但介导通道感知生理信号以及打开和关闭其门控方式的结构重排无法直接通过电学方法测量。在此,我们描述了一种基于用环境敏感荧光团对感兴趣的通道进行位点特异性标记的方法,该方法能够实时监测生物膜中离子通道的构象变化。