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. 2022:2507:389-404.
doi: 10.1007/978-1-0716-2368-8_21.

Detergent-Free Membrane Protein Purification Using SMA Polymer

Luke Broadbent #  1 , Peer Depping #  1 , Alexis Lodé #  1 , Afroditi Vaitsopoulou #  1 , David Hardy  1   2 , Hoor Ayub  1   3 , James Mitchell-White  4 , Ian D Kerr  4 , Alan D Goddard  1 , Roslyn M Bill  1 , Alice J Rothnie  5
Affiliations

Detergent-Free Membrane Protein Purification Using SMA Polymer

Luke Broadbent et al. Methods Mol Biol. 2022.

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.

Keywords: Detergent; Membrane proteins; Nanodisc; Polymer; Purification; SMALP; Solubilization.

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