通过取代半胱氨酸可及性方法（SCAM™）进行跨膜蛋白拓扑结构绘图：在脂质特异性膜蛋白拓扑发生中的应用
Transmembrane protein topology mapping by the substituted cysteine accessibility method (SCAM(TM)): application to lipid-specific membrane protein topogenesis.
作者信息
Bogdanov Mikhail, Zhang Wei, Xie Jun, Dowhan William
机构信息
Department of Biochemistry and Molecular Biology, University of Texas-Houston, Medical School, Houston, TX 77030, USA.
出版信息
Methods. 2005 Jun;36(2):148-71. doi: 10.1016/j.ymeth.2004.11.002.
Abstract
We provide an overview of lipid-dependent polytopic membrane protein topogenesis, with particular emphasis on Escherichia coli strains genetically altered in their lipid composition and strategies for experimentally determining the transmembrane organization of proteins. A variety of reagents and experimental strategies are described including the use of lipid mutants and thiol-specific chemical reagents to study lipid-dependent and host-specific membrane protein topogenesis by substituted cysteine site-directed chemical labeling. Employing strains in which lipid composition can be controlled temporally during membrane protein synthesis and assembly provides a means to observe dynamic changes in protein topology as a function of membrane lipid composition.
摘要
我们概述了脂质依赖性多跨膜蛋白的拓扑形成过程，特别强调了脂质组成发生基因改变的大肠杆菌菌株，以及通过实验确定蛋白质跨膜组织的策略。文中描述了多种试剂和实验策略，包括利用脂质突变体和硫醇特异性化学试剂，通过取代半胱氨酸定点化学标记来研究脂质依赖性和宿主特异性膜蛋白的拓扑形成。使用在膜蛋白合成和组装过程中脂质组成可随时间控制的菌株，为观察蛋白质拓扑结构随膜脂质组成变化的动态改变提供了一种方法。
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