Fiche publication
Date publication
janvier 2016
Journal
Biochemistry
Auteurs
Membres identifiés du Cancéropôle Est :
Dr WAGNER Renaud
Tous les auteurs :
Logez C, Damian M, Legros C, Dupré C, Guéry M, Mary S, Wagner R, M'Kadmi C, Nosjean O, Fould B, Marie J, Fehrentz JA, Martinez J, Ferry G, Boutin JA, Banères JL
Lien Pubmed
Résumé
G protein-coupled receptors (GPCRs) are integral membrane proteins that play a pivotal role in signal transduction. Understanding their dynamics is absolutely required to get a clear picture of how signaling proceeds. Molecular characterization of GPCRs isolated in detergents nevertheless stumbles over the deleterious effect of these compounds on receptor function and stability. We explored here the potential of a styrene-maleic acid polymer to solubilize receptors directly from their lipid environment. To this end, we used two GPCRs, the melatonin and ghrelin receptors, embedded in two membrane systems of increasing complexity, liposomes and membranes from Pichia pastoris. The styrene-maleic acid polymer was able, in both cases, to extract membrane patches of a well-defined size. GPCRs in SMA-stabilized lipid discs not only recognized their ligand but also transmitted a signal, as evidenced by their ability to activate their cognate G proteins and recruit arrestins in an agonist-dependent manner. Besides, the purified receptor in lipid discs undergoes all specific changes in conformation associated with ligand-mediated activation, as demonstrated in the case of the ghrelin receptor with fluorescent conformational reporters and compounds from distinct pharmacological classes. Altogether, these data highlight the potential of styrene-maleic stabilized lipid discs for analyzing the molecular bases of GPCR-mediated signaling in a well-controlled membrane-like environment.
Mots clés
Animals, CHO Cells, Cricetulus, GTP-Binding Proteins, chemistry, Humans, Lipids, chemistry, Liposomes, chemistry, Maleates, chemistry, Models, Molecular, Nanostructures, chemistry, Pichia, chemistry, Polystyrenes, chemistry, Receptors, Ghrelin, chemistry, Receptors, Melatonin, chemistry, Solubility
Référence
Biochemistry. 2016 Jan 12;55(1):38-48
