Pollock, NL, Rai, M, Simon, KS et al. (18 more authors) (2019) SMA-PAGE: A new method to examine complexes of membrane proteins using SMALP nano-encapsulation and native gel electrophoresis. BBA - Biomembranes, 1861 (8). pp. 1437-1445. ISSN 0005-2736
Abstract
Most membrane proteins function through interactions with other proteins in the phospholipid bilayer, the cytosol or the extracellular milieu. Understanding the molecular basis of these interactions is key to understanding membrane protein function and dysfunction. Here we demonstrate for the first time how a nano-encapsulation method based on styrene maleic acid lipid particles (SMALPs) can be used in combination with native gel electrophoresis to separate membrane protein complexes in their native state. Using four model proteins, we show that this separation method provides an excellent measure of protein quaternary structure, and that the lipid environment surrounding the protein(s) can be probed using mass spectrometry. We also show that the method is complementary to immunoblotting. Finally we show that intact membrane protein-SMALPs extracted from a band on a gel could be visualised using electron microscopy (EM). Taken together these results provide a novel and elegant method for investigating membrane protein complexes in a native state.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © 2019 Published by Elsevier B.V. This is an author produced version of a paper published in Biochimica et Biophysica Acta (BBA) - Biomembranes. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | Membrane protein; Nanoparticle; SMALP; Native PAGE; Protein complex |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Biological Sciences (Leeds) > School of Biomedical Sciences (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 23 May 2019 11:00 |
Last Modified: | 28 May 2020 00:38 |
Status: | Published |
Publisher: | Elsevier |
Identification Number: | 10.1016/j.bbamem.2019.05.011 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:146483 |
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Filename: Pollock Native PAGE April19 submission unmarked.pdf
Licence: CC-BY-NC-ND 4.0