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ToolsSwainsbury, David J. K., Scheidelaar, Stefan, Foster, Nicholas, van Grondelle, Rienk, Killian, J. Antoinette and Jones, Michael R. (2017) The effectiveness of styrene-maleic acid (SMA) copolymers for solubilisation of integral membrane proteins from SMA-accessible and SMA-resistant membranes. Biochimica et Biophysica Acta - Biomembranes, 1859 (10). pp. 2133-2143. ISSN 0005-2736
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Abstract
Solubilisation of biological lipid bilayer membranes for analysis of their protein complement has traditionally been carried out using detergents, but there is increasing interest in the use of amphiphilic copolymers such as styrene maleic acid (SMA) for the solubilisation, purification and characterisation of integral membrane proteins in the form of protein/lipid nanodiscs. Here we survey the effectiveness of various commercially-available formulations of the SMA copolymer in solubilising Rhodobacter sphaeroides reaction centres (RCs) from photosynthetic membranes. We find that formulations of SMA with a 2:1 or 3:1 ratio of styrene to maleic acid are almost as effective as detergent in solubilising RCs, with the best solubilisation by short chain variants (< 30 kDa weight average molecular weight). The effectiveness of 10 kDa 2:1 and 3:1 formulations of SMA to solubilise RCs gradually declined when genetically-encoded coiled-coil bundles were used to artificially tether normally monomeric RCs into dimeric, trimeric and tetrameric multimers. The ability of SMA to solubilise reaction centre-light harvesting 1 (RC-LH1) complexes from densely packed and highly ordered photosynthetic membranes was uniformly low, but could be increased through a variety of treatments to increase the lipid:protein ratio. However, proteins isolated from such membranes comprised clusters of complexes in small membrane patches rather than individual proteins. We conclude that short-chain 2:1 and 3:1 formulations of SMA are the most effective in solubilising integral membrane proteins, but that solubilisation efficiencies are strongly influenced by the size of the target protein and the density of packing of proteins in the membrane.
| Item Type: | Article |
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| Additional Information: | Funding Information: We thank Dr. Christine Moriscot from the Electron Microscopy platform of the Integrated Structural Biology of Grenoble (ISBG, UMI3265) for the collection of electron-microscopy data. This work used the platforms of the Grenoble Instruct centre (ISBG; UMS 3518 CNRS-CEA-UJF-EMBL) with support from FRISBI (ANR-10-INSB-05-02) and GRAL (ANR-10-LABX-49-01) within the Grenoble Partnership for Structural Biology (PSB). The electron microscope facility is supported by the Rhône-Alpes Region, the Fondation Recherche Medicale (FRM), the fonds FEDER, the Centre National de la Recherche Scientifique (CNRS), the CEA, the University of Grenoble, EMBL, and the GIS-Infrastrutures en Biologie Sante et Agronomie (IBISA). Publisher Copyright: © 2017 The Authors |
| Uncontrolled Keywords: | detergent,light harvesting,membrane protein,nanodisc,reaction centre,styrene—maleic acid,biophysics,biochemistry,cell biology ,/dk/atira/pure/subjectarea/asjc/1300/1304 |
| Faculty \ School: | Faculty of Science > School of Biological Sciences |
| UEA Research Groups: | Faculty of Science > Research Groups > Molecular Microbiology |
| Related URLs: | |
| Depositing User: | LivePure Connector |
| Date Deposited: | 17 Aug 2022 12:31 |
| Last Modified: | 09 Nov 2024 00:50 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/87368 |
| DOI: | 10.1016/j.bbamem.2017.07.011 |
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