SMA-PAGE: A new method to examine complexes of membrane proteins using SMALP nano-encapsulation and native gel electrophoresis


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.

Publication DOI:
Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences
College of Health & Life Sciences > Chronic and Communicable Conditions
Additional Information: © 2019, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International Funding: European Union under a Marie Skłodwska-Curie Initial Training Network FP7-PEOPLE-2012-ITN Grant Agreement Number 316630 CAS IDP, BBSRC Tools and Resources Development Fund grant number BB/P027482/1.
Uncontrolled Keywords: Membrane protein,Nanoparticle,Native PAGE,Protein complex,SMALP,Biophysics,Biochemistry,Cell Biology
Publication ISSN: 1879-2642
Last Modified: 10 Jun 2024 07:34
Date Deposited: 19 Aug 2019 09:30
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Related URLs: https://linking ... 005273619301087 (Publisher URL)
http://www.scop ... tnerID=8YFLogxK (Scopus URL)
PURE Output Type: Article
Published Date: 2019-08-01
Published Online Date: 2019-05-28
Accepted Date: 2019-05-19
Authors: Pollock, Naomi L.
Rai, Megha
Simon, Kailene S.
Hesketh, Sophie J.
Teo, Alvin C.k.
Parmar, Mayuriben
Sridhar, Pooja
Collins, Richard
Lee, Sarah C.
Stroud, Zoe N.
Bakker, Saskia E.
Muench, Stephen P.
Barton, C. Howard
Hurlbut, Gregory
Roper, David I.
Smith, Corinne J.i.
Knowles, Timothy J.
Spickett, Corinne M. (ORCID Profile 0000-0003-4054-9279)
East, J. Malcolm
Postis, Vincent
Dafforn, Tim R.

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