Membrane Protein Solubilization英文文献.pdfVIP

Chapter 13 Membrane Protein Solubilization Katia Duquesne and James N. Sturgis 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. Key words: Cytoplasmic membrane, detergent, membrane proteins, purification, titration 1. Introduction The solubilization of a membrane protein from a biological sample often involves a series of related steps. It is hard to estab- lish a method without examining the various steps involved and the use to which the solubilized protein will be put. Here, we describe an approach to optimising the solubilization of your membrane protein of interest; we presume that you have some method of specifically detecting the presence of this protein and some measure of its activity or function. While the main theme of this chapter is the solubilization of membrane protei