Detergent-Free Membrane Protein Purification Using SMA Polymer

Luke Broadbent; Peer Depping; Alexis Lodé; Afroditi Vaitsopoulou; David Hardy; Hoor Ayub; James Mitchell-White; Ian D Kerr; Alan D Goddard; Roslyn M Bill; Alice J Rothnie

doi:10.1007/978-1-0716-2368-8_21

Detergent-Free Membrane Protein Purification Using SMA Polymer

Luke Broadbent, Peer Depping, Alexis Lodé, Afroditi Vaitsopoulou, David Hardy, Hoor Ayub, James Mitchell-White, Ian D Kerr, Alan D Goddard, Roslyn M Bill, Alice J Rothnie

School of Health and Care

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

2 Citations (Scopus)

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. [Abstract copyright: © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.]

Original language	English
Title of host publication	Heterologous Expression of Membrane Proteins. Methods in Molecular Biology
Editors	I Mus-Veteau
Publisher	Humana Press
Pages	389-404
Number of pages	16
Volume	2507
ISBN (Electronic)	978-1-0716-2368-8
ISBN (Print)	978-1-0716-2367-1
DOIs	https://doi.org/10.1007/978-1-0716-2368-8_21
Publication status	Published - 1 Jul 2022

Publication series

Name	Methods in molecular biology (Clifton, N.J.)
Publisher	Humana Press
ISSN (Print)	1064-3745

Bibliographical note

Funding Information:
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

Publisher Copyright:
© 2022, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords

Polystyrenes - chemistry
Nanodisc
Detergent
Polymer
SMALP
Purification
Membrane proteins
Solubilization
Chromatography, Affinity
Membrane Proteins - chemistry
Membranes
Polymers - chemistry
Lipid Bilayers - chemistry

Access to Document

10.1007/978-1-0716-2368-8_21

Cite this

Broadbent, L., Depping, P., Lodé, A., Vaitsopoulou, A., Hardy, D., Ayub, H., Mitchell-White, J., Kerr, I. D., Goddard, A. D., Bill, R. M., & Rothnie, A. J. (2022). Detergent-Free Membrane Protein Purification Using SMA Polymer. In I. Mus-Veteau (Ed.), Heterologous Expression of Membrane Proteins. Methods in Molecular Biology (Vol. 2507, pp. 389-404). (Methods in molecular biology (Clifton, N.J.)). Humana Press. https://doi.org/10.1007/978-1-0716-2368-8_21

Detergent-Free Membrane Protein Purification Using SMA Polymer. / Broadbent, Luke; Depping, Peer; Lodé, Alexis et al.
Heterologous Expression of Membrane Proteins. Methods in Molecular Biology. ed. / I Mus-Veteau. Vol. 2507 Humana Press, 2022. p. 389-404 (Methods in molecular biology (Clifton, N.J.)).

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

Broadbent, L, Depping, P, Lodé, A, Vaitsopoulou, A, Hardy, D, Ayub, H, Mitchell-White, J, Kerr, ID, Goddard, AD, Bill, RM & Rothnie, AJ 2022, Detergent-Free Membrane Protein Purification Using SMA Polymer. in I Mus-Veteau (ed.), Heterologous Expression of Membrane Proteins. Methods in Molecular Biology. vol. 2507, Methods in molecular biology (Clifton, N.J.), Humana Press, pp. 389-404. https://doi.org/10.1007/978-1-0716-2368-8_21

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Detergent-Free Membrane Protein Purification Using SMA Polymer

Abstract

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Keywords

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