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

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., East, J. Malcolm, Postis, Vincent and Dafforn, Tim R. (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.

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.

Publication DOI: https://doi.org/10.1016/j.bbamem.2019.05.011
Divisions: Life & Health Sciences > Biosciences
Life & Health Sciences
Life & Health Sciences > Chronic and Communicable Conditions
Additional Information: © 2019, Elsevier. Licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International http://creativecommons.org/licenses/by-nc-nd/4.0/ 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
Full Text Link:
Related URLs: https://linking ... 005273619301087 (Publisher URL)
http://www.scop ... tnerID=8YFLogxK (Scopus URL)
Published Date: 2019-08-01
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. ( 0000-0003-4054-9279)
East, J. Malcolm
Postis, Vincent
Dafforn, Tim R.

Download

[img]

Version: Accepted Version

Access Restriction: Restricted to Repository staff only until 28 May 2020.

License: Creative Commons Attribution Non-commercial No Derivatives


Export / Share Citation


Statistics

Additional statistics for this record