Protein reconstitution into freestanding planar lipid membranes for electrophysiological characterization
Protein reconstitution into freestanding planar lipid membranes for electrophysiological characterization
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DOI：
10.1038/nprot.2015.003
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发表时间：
2015-01-01
期刊：
影响因子：
14.8
通讯作者：
Winterhalter, Mathias
中科院分区：
文献类型：
--
作者：
Gutsmann, Thomas;Heimburg, Thomas;Winterhalter, Mathias
The reconstitution of channel-forming proteins into planar lipid bilayers enables their functional characterization at very low (sometimes below attomolar) concentrations. We describe the three main approaches used in our laboratories (the Mueller-Rudin technique, in which the bilayers contain an organic solvent, the Montal-Mueller or solvent-free technique, and a method for membrane reconstitution via liposome formation), and we discuss their respective advantages and limitations. Despite the differences in the reconstitution procedures, subsequent protein characterization is based on the same electrophysiological technique. A transmembrane electric field is applied, inducing an ion current and allowing conclusions to be drawn on apparent pore sizes, or suggesting functional properties such as channel opening and closing upon ligand binding, pH-induced conformational changes, ion selectivity or substrate specificity.
