Journal
PROTEIN EXPRESSION AND PURIFICATION
Volume 86, Issue 1, Pages 12-20Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2012.08.016
Keywords
Membrane proteins; Extraction efficiency; Detergents; Surfactants; Mass spectrometry
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Funding
- National Science Foundation [0923184]
- Direct For Biological Sciences
- Div Of Biological Infrastructure [0923184] Funding Source: National Science Foundation
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Generating stable conditions for membrane proteins after extraction from their lipid bilayer environment is essential for subsequent characterization. Detergents are the most widely used means to obtain this stable environment; however, different types of membrane proteins have been found to require detergents with varying properties for optimal extraction efficiency and stability after extraction. The extraction profiles of several detergent types have been examined for membranes isolated from bacteria and yeast, and for a set of recombinant target proteins. The extraction efficiencies of these detergents increase at higher concentrations, and were shown to correlate with their respective CMC values. Two alkyl sugar detergents, octyl-beta-D-glucoside (OG) and 5-cyclohexyl-1-pentyl-beta-D-maltoside (Cymal-5), and a zwitter-ionic surfactant, N-decylphosphocholine (Fos-choline-10), were generally effective in the extraction of a broad range of membrane proteins. However, certain detergents were more effective than others in the extraction of specific classes of integral membrane proteins, offering guidelines for initial detergent selection. The differences in extraction efficiencies among this small set of detergents supports the value of detergent screening and optimization to increase the yields of targeted membrane proteins. (c) 2012 Elsevier Inc. All rights reserved.
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