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

Research output: Contribution to journalArticle

Authors

  • Megha Rai
  • Kailene S Simon
  • Sophie J Hesketh
  • Alvin C K Teo
  • Mayuriben Parmar
  • Pooja Sridhar
  • Richard Collins
  • Zoe N Stroud
  • Saskia E Bakker
  • Stephen P Muench
  • C Howard Barton
  • Gregory Hurlbut
  • David I Roper
  • Corinne J I Smith
  • Corinne M Spickett
  • J Malcolm East
  • Vincent L G Postis

Colleges, School and Institutes

External organisations

  • Centre for Biological Sciences, University of Southampton
  • University of Massachusetts Medical School, USA; Rare and Neurologic Diseases Research, Sanofi, MA, USA.
  • School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, UK; School of Biology, University of Manchester, UK.
  • School of Life Sciences, University of Warwick
  • University of Leeds
  • School of Clinical and Applied Sciences, Leeds Beckett University, UK.
  • Rare and Neurologic Diseases Research, Sanofi, MA, USA.
  • School of Life and Health Sciences, Aston University

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.

Bibliographic note

Copyright © 2019. Published by Elsevier B.V.

Details

Original languageEnglish
Pages (from-to)1437-1445
Number of pages9
JournalBiochimica et Biophysica Acta (BBA) - Biomembranes
Volume1861
Issue number8
Early online date28 May 2019
Publication statusPublished - 1 Aug 2019

Keywords

  • Membrane protein, Nanoparticle, Native PAGE, Protein complex, SMALP

ASJC Scopus subject areas