Membrane protein extraction and purification using styrene-maleic acid (SMA) copolymer: effect of variations in polymer structure

The use of styrene-maleic acid (SMA) copolymers to extract and purify transmembrane proteins, while retaining their native bilayer environment, overcomes many of the disadvantages associated with conventional detergent-based procedures. This approach has huge potential for the future of membrane pro...

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Bibliographic Details
Published in:	Biochemical journal Vol. 473; no. 23; p. 4349
Main Authors:	Morrison, Kerrie A, Akram, Aneel, Mathews, Ashlyn, Khan, Zoeya A, Patel, Jaimin H, Zhou, Chumin, Hardy, David J, Moore-Kelly, Charles, Patel, Roshani, Odiba, Victor, Knowles, Tim J, Javed, Masood-Ul-Hassan, Chmel, Nikola P, Dafforn, Timothy R, Rothnie, Alice J
Format:	Journal Article
Language:	English
Published:	England 01-12-2016
Subjects:	Carrier Proteins - chemistry Carrier Proteins - isolation & purification Cell Cycle Proteins - chemistry Cell Cycle Proteins - isolation & purification Escherichia coli Proteins - chemistry Escherichia coli Proteins - isolation & purification Maleates - chemistry Membrane Proteins - chemistry Membrane Proteins - isolation & purification Polystyrenes - chemistry Solubility polymer protein purification membrane proteins solubilisation SMA SMALP
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Summary:	The use of styrene-maleic acid (SMA) copolymers to extract and purify transmembrane proteins, while retaining their native bilayer environment, overcomes many of the disadvantages associated with conventional detergent-based procedures. This approach has huge potential for the future of membrane protein structural and functional studies. In this investigation, we have systematically tested a range of commercially available SMA polymers, varying in both the ratio of styrene and maleic acid and in total size, for the ability to extract, purify and stabilise transmembrane proteins. Three different membrane proteins (BmrA, LeuT and ZipA), which vary in size and shape, were used. Our results show that several polymers, can be used to extract membrane proteins, comparably to conventional detergents. A styrene:maleic acid ratio of either 2:1 or 3:1, combined with a relatively small average molecular mass (7.5-10 kDa), is optimal for membrane extraction, and this appears to be independent of the protein size, shape or expression system. A subset of polymers were taken forward for purification, functional and stability tests. Following a one-step affinity purification, SMA 2000 was found to be the best choice for yield, purity and function. However, the other polymers offer subtle differences in size and sensitivity to divalent cations that may be useful for a variety of downstream applications.
ISSN:	1470-8728
DOI:	10.1042/bcj20160723