Martinac Boris
Victor Chang Cardiac Research Institute, Lowy Packer Building, Darlinghurst, NSW 2010, Australia; St Vincent's Clinical School, University of New South Wales, Darlinghurst, NSW 2010, Australia.
Prog Biophys Mol Biol. 2017 Nov;130(Pt B):192-197. doi: 10.1016/j.pbiomolbio.2017.06.014. Epub 2017 Jun 23.
Different types of fluorescence spectroscopy approaches have over the last two decades become important techniques in studies of ion channel structure and dynamics. Many fluorescence methods have been used to examine a huge variety of ion channels. Any fluorescence study of ion channels requires the presence of fluorophores, which may be intrinsic to the channel protein, attached either extrinsically to the protein, or be simply located nearby the channel to monitor local conditions such as for many of the ion-sensitive dyes. Many ion channel studies utilize protein-bound or intrinsic protein fluorophores. Single-molecule Förster resonance energy transfer (smFRET) spectroscopy has been particularly useful in gaining detailed structural information for multimeric membrane proteins including ion channels. This technique presents a major advancement in studies of structural dynamics of these membrane proteins. Although it has required different approaches to protein labelling, control of the protein state, as well as careful analysis of the orientations, geometries, and number of fluorescent probes, the smFRET methodology provides an excellent tool for studying the structure of ion channels.
在过去二十年中，不同类型的荧光光谱方法已成为研究离子通道结构和动力学的重要技术。许多荧光方法已被用于检测各种各样的离子通道。任何关于离子通道的荧光研究都需要有荧光团，荧光团可能是通道蛋白固有的，也可能是外在附着于蛋白的，或者只是位于通道附近以监测局部条件，例如许多离子敏感染料的情况。许多离子通道研究利用与蛋白结合的或蛋白固有的荧光团。单分子荧光共振能量转移（smFRET）光谱在获取包括离子通道在内的多聚体膜蛋白的详细结构信息方面特别有用。这项技术在这些膜蛋白的结构动力学研究中取得了重大进展。尽管它需要不同的蛋白质标记方法、对蛋白质状态的控制以及对荧光探针的方向、几何形状和数量进行仔细分析，但smFRET方法为研究离子通道的结构提供了一个极好的工具。
