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

Pollock, NL and Rai, M and Simon, KS and Hesketh, SJ and Teo, ACK and Parmar, M and Sridhar, P and Collins, R and Lee, SC and Stroud, ZN and Bakker, SE and Muench, SP and Barton, CH and Hurlbut, G and Roper, DI and Smith, CJI and Knowles, TJ and Spickett, CM and East, JM and Postis, V and Dafforn, TR (2019) SMA-PAGE: A new method to examine complexes of membrane proteins using SMALP nano-encapsulation and native gel electrophoresis. Biochim Biophys Acta Biomembr, 1861 (8). pp. 1437-1445. ISSN 1879-2642 DOI: https://doi.org/10.1016/j.bbamem.2019.05.011

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.

Identification Number:	https://doi.org/10.1016/j.bbamem.2019.05.011
Status:	Published
Refereed:	Yes
Uncontrolled Keywords:	Membrane protein, Nanoparticle, Native PAGE, Protein complex, SMALP,
Depositing User (symplectic)	Deposited by Postis, Vincent
Date Deposited:	11 Jun 2019 13:01
Last Modified:	11 Jul 2024 15:08
Item Type:	Article

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

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