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

Morrison, Kerrie A., Akram, Aneel, Mathews, Ashlyn, Khan, Zoeya A., Patel, Jaimin H., Zhou, Chumin, Hardy, David John, Moore-Kelly, Charles, Patel, Roshani, Odiba, Victor, Knowles, Tim, Javed, Masood-ul-Hassan, Chmel, Nikola P., Dafforn, Timothy R. and Rothnie, Alice Jane (2016). Membrane protein extraction and purification using styrene-maleic acid (SMA) co-polymer:effect of variations in polymer structure. Biochemical Journal, 473 (23), pp. 4349-4360.

Abstract

The use of styrene maleic acid (SMA) co-polymers to extract and purify transmembrane proteins, whilst 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 to 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 weight (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.

Publication DOI: https://doi.org/10.1042/BCJ20160723
Dataset DOI: https://doi.org/10.17036/ff13833e-68c5-449f-82f2-4f07363fc55d
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Life & Health Sciences > Cellular and Molecular Biomedicine
Additional Information: Biochemical Journal. This is not the final peer-reviewed Version of Record, which is available on http://dx.doi.org/10.1042/BCJ20160723
Uncontrolled Keywords: Biochemistry,Molecular Biology,Cell Biology
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Related URLs: http://www.scop ... tnerID=8YFLogxK (Scopus URL)
Published Date: 2016-12-01
Authors: Morrison, Kerrie A.
Akram, Aneel
Mathews, Ashlyn
Khan, Zoeya A.
Patel, Jaimin H.
Zhou, Chumin
Hardy, David John
Moore-Kelly, Charles
Patel, Roshani
Odiba, Victor
Knowles, Tim
Javed, Masood-ul-Hassan
Chmel, Nikola P.
Dafforn, Timothy R.
Rothnie, Alice Jane ( 0000-0002-4259-7015)

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