Referenced in: none

Automatically associated channels: Kv10.1

Title: A high-throughput differential filtration assay to screen and select detergents for membrane proteins.

Authors: James M Vergis, Michael D Purdy, Michael C Wiener

Journal, date & volume: Anal. Biochem., 2010 Dec 1 , 407, 1-11

PubMed link: http://www.ncbi.nlm.nih.gov/pubmed/20667442

Abstract
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.