使用荧光共振能量转移光谱法测量的MscL通道门控过程中涉及的构象变化。
Conformational changes involved in MscL channel gating measured using FRET spectroscopy.
作者信息
Corry Ben, Rigby Paul, Liu Zhen-Wei, Martinac Boris
机构信息
School of Biomedical, Biomolecular and Chemical Science, The University of Western Australia, Crawley, Australia.
出版信息
Biophys J. 2005 Dec;89(6):L49-51. doi: 10.1529/biophysj.105.072009. Epub 2005 Sep 30.
Abstract
We demonstrate that fluorescence resonance energy transfer spectroscopy is a powerful tool for in situ structural analysis of multimeric membrane proteins by measuring the conformational changes involved in gating the mechanosensitive ion channel of large conductance. Ensemble analysis is used to analyze the intensity of light emitted by AlexaFluor-labeled cysteine mutants reconstituted into artificial liposomes before and after acceptor photobleaching. The diameter of the protein is found to increase by 16 A upon channel activation.
摘要
我们证明，荧光共振能量转移光谱法是用于多聚体膜蛋白原位结构分析的强大工具，通过测量参与大电导机械敏感离子通道门控的构象变化来实现。整体分析用于分析受体光漂白前后重构到人工脂质体中的AlexaFluor标记的半胱氨酸突变体发出的光强度。发现通道激活后蛋白质直径增加了16埃。
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