An improved tripod amphiphile for membrane protein solubilization
An improved tripod amphiphile for membrane protein solubilization
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DOI：
10.1110/ps.9.12.2518
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发表时间：
2000-12-01
期刊：
影响因子：
8
通讯作者：
Gellman, SH
中科院分区：
文献类型：
--
作者：
Yu, SM;McQuade, DT;Gellman, SH
内在膜蛋白代表了由活生物体产生的蛋白质的大部分，并且执行许多关键功能。膜蛋白的结构和功能表征通常需要从天然脂质双层中提取它们，并用小的合成两亲物（例如去污剂）溶解。我们描述了一种具有独特的两亲性结构的小分子的发展，一种“三脚架两亲物”，它可以溶解细菌视紫红质（BR）和牛视紫红质（Rho）。这种两亲物的极性部分含有酰胺和氧化胺：在这个极性段的小变化被发现有深远的影响蛋白质的溶解性能。最佳的三脚架两亲物从天然膜环境中提取BR和Rho，并在脱脂后数周内保持每种蛋白质以单体天然样形式存在。三脚架两亲物被设计成比常规洗涤剂显示更大的构象刚性，其长期目标是促进膜蛋白结晶。这里报告的结果是朝着这一最终目标迈出的重要一步。
Intrinsic membrane proteins represent a large fraction of the proteins produced by living organisms and perform many crucial functions. Structural and functional characterization of membrane proteins generally requires that they be extracted from the native lipid bilayer and solubilized with a small synthetic amphiphile, for example, a detergent. We describe the development of a small molecule with a distinctive amphiphilic architecture, a "tripod amphiphile," that solubilizes both bacteriorhodopsin (BR) and bovine rhodopsin (Rho). The polar portion of this amphiphile contains an amide and an amine-oxide: small variations in this polar segment are found to have profound effects on protein solubilization properties. The optimal tripod amphiphile extracts both BR and Rho from the native membrane environments and maintains each protein in a monomeric native-like form for several weeks after delipidation. Tripod amphiphiles are designed to display greater conformational rigidity than conventional detergents, with the long-range goal of promoting membrane protein crystallization. The results reported here represent an important step toward that ultimate goal.