Broadbent, Luke, Depping, Peer, Lodé, Alexis, Vaitsopoulou, Afroditi, Hardy, David, Ayub, Hoor, Mitchell-White, James, Kerr, Ian D, Goddard, Alan D, Bill, Roslyn M and Rothnie, Alice J (2022). Detergent-Free Membrane Protein Purification Using SMA Polymer. IN: Methods in Molecular Biology. Mus-Veteau, I. (ed.) Methods in Molecular Biology, 2507 . Springer.
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.
Publication DOI: | https://doi.org/10.1007/978-1-0716-2368-8_21 |
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Divisions: | College of Health & Life Sciences > School of Biosciences College of Health & Life Sciences College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine Aston University (General) |
Funding Information: | 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/R02152 |
Additional Information: | Copyright © Springer Nature B.V. 2022. The final publication is available at Springer via https://doi.org/10.1007/978-1-0716-2368-8_21 |
Uncontrolled Keywords: | Chromatography, Affinity,Lipid Bilayers/chemistry,Membrane Proteins/chemistry,Membranes,Polymers/chemistry,Polystyrenes/chemistry,Detergent,Purification,Polymer,Solubilization,Membrane proteins,Nanodisc,SMALP,Genetics,Molecular Biology |
ISBN: | 978-1-0716-2367-1, 978-1-0716-2368-8 |
Last Modified: | 04 Nov 2024 09:47 |
Date Deposited: | 11 Jul 2024 14:40 |
Full Text Link: | |
Related URLs: |
https://link.sp ... -0716-2368-8_21
(Publisher URL) http://www.scop ... tnerID=8YFLogxK (Scopus URL) |
PURE Output Type: | Chapter (peer-reviewed) |
Published Date: | 2022-07-01 |
Authors: |
Broadbent, Luke
Depping, Peer ( 0000-0002-5694-2393) Lodé, Alexis Vaitsopoulou, Afroditi Hardy, David Ayub, Hoor Mitchell-White, James Kerr, Ian D Goddard, Alan D ( 0000-0003-4950-7470) Bill, Roslyn M ( 0000-0003-1331-0852) Rothnie, Alice J ( 0000-0002-4259-7015) |