Article
Received: 1 September 2008; Accepted: 27 November 2008
10.1002/pro.55 About DOI
Engineering a G protein-coupled receptor for structural studies: Stabilization of the BLT1 receptor ground state |
| Aimée Martin 1, Marjorie Damian 1, Michel Laguerre 2, Joseph Parello 3, Bernard Pucci 4, Laurence Serre 5 a, Sophie Mary 1, Jacky Marie 1, Jean-Louis Banères 1 * |
| 1Institut des Biomolécules Max Mousseron (IBMM), UMR 5247 CNRS Universités Montpellier I et II, Faculté de Pharmacie, 15 Av. Ch. Flahault, BP14491, 34093 Montpellier Cedex 5, France 2Institut Européen de Chimie et Biologie, UMR 5248 CNRS, 2 rue Robert-Escarpit, 33607 Pessac Cedex, France 3Department of Pharmacology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-6600, USA 4Laboratoire de Chimie Bioorganique et des Systèmes Moléculaires Vectoriels, Université d'Avignon et des Pays du Vaucluse, Faculté des Sciences, 33 Rue Louis Pasteur, F-84000 Avignon, France 5Laboratoire des Protéines Membranaires, Institut de Biologie Structurale Jean-Pierre Ebel CEA/CNRS/UJF, 41, rue Jules Horowitz, 38027 Grenoble cedex 01, France |
| email: Jean-Louis Banères (baneres@univ-montp1.fr) |
*Correspondence to Jean-Louis Banères, IBMM, Faculté de Pharmacie Bât. K, 15 Av. Charles Flahault, BP14491, 34093 Montpellier Cedex 5, France
aCurrent address: ESRF, Partner for Structural Biology, 6, rue Jules Horowitz, 38042 Grenoble, France
Funded by:
Ministère de la Recherche; Grant Number: ANR 06-BLAN-0087 l'Association pour la Recherche sur le Cancer (subvention libre n°3159) CNRS
| Keywords |
| GPCR G protein activation, structure stability |
| Abstract |
| Structural characterization of membrane proteins is hampered by their instability in detergent solutions. We modified here a G protein-coupled receptor, the BLT1 receptor of leukotriene B4, to stabilize it in vitro. For this, we introduced a metal-binding site connecting the third and sixth transmembrane domains of the receptor. This modification was intended to restrain the activation-associated relative movement of these helices that results in a less stable packing in the isolated receptor. The modified receptor binds its agonist with low-affinity and can no longer trigger G protein activation, indicating that it is stabilized in its ground state conformation. Of importance, the modified BLT1 receptor displays an increased temperature-, detergent-, and time-dependent stability compared with the wild-type receptor. These data indicate that stabilizing the ground state of this GPCR by limiting the activation-associated movements of the transmembrane helices is a way to increase its stability in detergent solutions; this could represent a forward step on the way of its crystallization. |
Received: 1 September 2008; Accepted: 27 November 2008
| Digital Object Identifier (DOI) |
10.1002/pro.55 About DOI
