A high-throughput differential filtration assay to screen and select detergents for membrane proteins
A high-throughput differential filtration assay to screen and select detergents for membrane proteins
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DOI：
10.1016/j.ab.2010.07.019
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发表时间：
2010-12-01
影响因子：
2.9
通讯作者：
Wiener, Michael C.
中科院分区：
文献类型：
--
作者：
Vergis, James M.;Purdy, Michael D.;Wiener, Michael C.
Structural studies on integral membrane proteins are routinely performed on protein-detergent complexes (PDCs) consisting of purified protein solubilized in a particular detergent. Of all the membrane protein crystal structures solved to date, a subset of only four detergents has been used in more than half of these structures. Unfortunately, many membrane proteins are not well behaved in these four detergents and/or fail to yield well-diffracting crystals. Identification of detergents that maintain the solubility and stability of a membrane protein is a critical step and can be a lengthy and "protein-expensive" process. We have developed an assay that characterizes the stability and size of membrane proteins exchanged into a panel of 94 commercially available and chemically diverse detergents. This differential filtration assay (DFA), using a set of filtered microplates, requires sub-milligram quantities of purified protein and small quantities of detergents and other reagents and is performed in its entirety in several hours. (C) 2010 Elsevier Inc. All rights reserved.
