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Detergent-Free Membrane Protein Purification Using SMA Polymer

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Heterologous Expression of Membrane Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2507))

Abstract

One of the big challenges for the study of structure and function of membrane proteins is the need to extract them from the membrane. Traditionally this was achieved using detergents which disrupt the membrane and form a micelle around the protein, but this can cause issues with protein function and/or stability. In 2009 an alternative approach was reported, using styrene maleic acid (SMA) copolymer to extract small discs of lipid bilayer encapsulated by the polymer and termed SMALPs (SMA lipid particles). Since then this approach has been shown to work for a range of different proteins from many different expression systems. It allows the extraction and purification of a target protein while maintaining a lipid bilayer environment. Recently this has led to several new high-resolution structures and novel insights to function. As with any method there are some limitations and issues to be aware of. Here we describe a standard protocol for preparation of the polymer and its use for membrane protein purification, and also include details of typical challenges that may be encountered and possible ways to address those.

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Acknowledgements

We are grateful for funding from the European Union’s Horizon 2020 research and innovation programme under Marie Sklodowska-Curie grant agreement No. 847,419 (MemTrain) as well as the ERACoBioTech MeMBrane project and BBSRC (BB/R02152X/1) to A.D.G, A.J.R and R.M.B. Also, BBSRC CASE studentship (BB/L015846/1) and BBSRC grant (BB/S008160/1) to A.J.R., and BBSRC grant (BB/S001611/1) to I.D.K.

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  1. Luke Broadbent, Peer Depping, Alexis Lodé and Afroditi Vaitsopoulou contributed equally.

Authors and Affiliations

  1. College of Health & Life Sciences, Aston University, Birmingham, UK

    Luke Broadbent, Peer Depping, Alexis Lodé, Afroditi Vaitsopoulou, David Hardy, Hoor Ayub, Alan D. Goddard, Roslyn M. Bill & Alice J. Rothnie

  2. School of Biosciences, University of Birmingham, Birmingham, UK

    David Hardy

  3. Faculty of Health & Life Sciences, Coventry University, Coventry, UK

    Hoor Ayub

  4. School of Life Sciences, University of Nottingham Medical School, Queen’s Medical Centre, Nottingham, UK

    James Mitchell-White & Ian D. Kerr

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  1. Université Côte d’Azur, CNRS UMR7275, Institut de Pharmacologie Moléculaire et Cellulaire, Valbonne, France

    Isabelle Mus-Veteau

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Broadbent, L. et al. (2022). Detergent-Free Membrane Protein Purification Using SMA Polymer. In: Mus-Veteau, I. (eds) Heterologous Expression of Membrane Proteins. Methods in Molecular Biology, vol 2507. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2368-8_21

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  • DOI: https://doi.org/10.1007/978-1-0716-2368-8_21

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