Protocol to test the utility of detergents for E. coli membrane protein extraction and delipidation

STAR Protoc. 2023 Mar 17;4(2):102146. doi: 10.1016/j.xpro.2023.102146. Online ahead of print.

Abstract

We present a protocol to evaluate the utility of detergents for purification and delipidation of E. coli membrane proteins. We determine the critical aggregation concentration of detergents. Furthermore, we compare the ability of detergents to extract membrane proteins and to maintain protein-lipid interactions during purification. The protocol describes steps for isolating and delipidating membrane proteins from E. coli membranes by extraction and affinity purification using detergents. The protocol does not enable an absolute quantification of purification outcomes. For complete details on the use and execution of this protocol, please refer to Urner et al.1.

Keywords: Mass Spectrometry; Model Organisms; Molecular/Chemical Probes; Protein Biochemistry; Protein Expression and Purification.