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膜蛋白增溶

Membrane protein solubilization.

作者信息

Duquesne Katia, Sturgis James N

机构信息

Institut de Biologie Structurale et Microbiologie, CNRS and Aix Marseille Université, France.

出版信息

Methods Mol Biol. 2010;601:205-17. doi: 10.1007/978-1-60761-344-2_13.

Abstract

A critical step in any in vitro analysis of membrane proteins is the solubilization of the membrane to extract the protein of interest in an active form to obtain an aqueous solution containing the membrane protein complexed with detergents and lipids in a form suitable for purification and further analysis. This process is particularly delicate as the aim is to maximally disrupt the lipid components of the membrane while putting the protein components in an un-natural detergent environment without perturbing them. Looked at this way, it is remarkable that it ever works. Although the process is difficult and hard to master, an increasing number of membrane proteins have been successfully solubilized in active forms, allowing some general principles to be established that we illustrate in the method developed in this chapter.

摘要

在对膜蛋白进行任何体外分析时,关键的一步是使膜溶解,以活性形式提取目标蛋白,从而获得一种水溶液,其中膜蛋白与去污剂和脂质形成复合物,其形式适合于纯化和进一步分析。这个过程特别微妙,因为目标是最大程度地破坏膜的脂质成分,同时将蛋白质成分置于非天然的去污剂环境中而不干扰它们。从这个角度来看,它竟然能够成功实在令人惊讶。尽管这个过程困难且难以掌握,但越来越多的膜蛋白已成功以活性形式溶解,这使得一些通用原则得以确立,我们将在本章所开发的方法中进行阐述。

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