Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid

Summary: The β2-adrenoceptor (β2AR) is a well-established target in asthma and a prototypical G protein-coupled receptor for biophysical studies. Solubilization of membrane proteins has classically involved the use of detergents. However, the detergent environment differs from the native membrane en...

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Autores principales:	Clare.R. Harwood, David A. Sykes, Bradley L. Hoare, Franziska M. Heydenreich, Romez Uddin, David R. Poyner, Stephen J. Briddon, D.B. Veprintsev
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Biophysical chemistry Membranes Protein Science Q
Acceso en línea:	https://doaj.org/article/8e54c20c4bce4994832b97948f5cb7d0
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spelling	oai:doaj.org-article:8e54c20c4bce4994832b97948f5cb7d02021-11-18T04:51:09ZFunctional solubilization of the β2-adrenoceptor using diisobutylene maleic acid2589-004210.1016/j.isci.2021.103362https://doaj.org/article/8e54c20c4bce4994832b97948f5cb7d02021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S258900422101333Xhttps://doaj.org/toc/2589-0042Summary: The β2-adrenoceptor (β2AR) is a well-established target in asthma and a prototypical G protein-coupled receptor for biophysical studies. Solubilization of membrane proteins has classically involved the use of detergents. However, the detergent environment differs from the native membrane environment and often destabilizes membrane proteins. Use of amphiphilic copolymers is a promising strategy to solubilize membrane proteins within their native lipid environment in the complete absence of detergents. Here we show the isolation of the β2AR in the polymer diisobutylene maleic acid (DIBMA). We demonstrate that β2AR remains functional in the DIBMA lipid particle and shows improved thermal stability compared with the n-dodecyl-β-D-maltopyranoside detergent-solubilized β2AR. This unique method of extracting β2AR offers significant advantages over previous methods routinely employed such as the introduction of thermostabilizing mutations and the use of detergents, particularly for functional biophysical studies.Clare.R. HarwoodDavid A. SykesBradley L. HoareFranziska M. HeydenreichRomez UddinDavid R. PoynerStephen J. BriddonD.B. VeprintsevElsevierarticleBiophysical chemistryMembranesProteinScienceQENiScience, Vol 24, Iss 12, Pp 103362- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biophysical chemistry Membranes Protein Science Q
spellingShingle	Biophysical chemistry Membranes Protein Science Q Clare.R. Harwood David A. Sykes Bradley L. Hoare Franziska M. Heydenreich Romez Uddin David R. Poyner Stephen J. Briddon D.B. Veprintsev Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid
description	Summary: The β2-adrenoceptor (β2AR) is a well-established target in asthma and a prototypical G protein-coupled receptor for biophysical studies. Solubilization of membrane proteins has classically involved the use of detergents. However, the detergent environment differs from the native membrane environment and often destabilizes membrane proteins. Use of amphiphilic copolymers is a promising strategy to solubilize membrane proteins within their native lipid environment in the complete absence of detergents. Here we show the isolation of the β2AR in the polymer diisobutylene maleic acid (DIBMA). We demonstrate that β2AR remains functional in the DIBMA lipid particle and shows improved thermal stability compared with the n-dodecyl-β-D-maltopyranoside detergent-solubilized β2AR. This unique method of extracting β2AR offers significant advantages over previous methods routinely employed such as the introduction of thermostabilizing mutations and the use of detergents, particularly for functional biophysical studies.
format	article
author	Clare.R. Harwood David A. Sykes Bradley L. Hoare Franziska M. Heydenreich Romez Uddin David R. Poyner Stephen J. Briddon D.B. Veprintsev
author_facet	Clare.R. Harwood David A. Sykes Bradley L. Hoare Franziska M. Heydenreich Romez Uddin David R. Poyner Stephen J. Briddon D.B. Veprintsev
author_sort	Clare.R. Harwood
title	Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid
title_short	Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid
title_full	Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid
title_fullStr	Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid
title_full_unstemmed	Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid
title_sort	functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/8e54c20c4bce4994832b97948f5cb7d0
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Functional solubilization of the β2-adrenoceptor using diisobutylene maleic acid

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