Leukotriene BLT2 receptor monomers activate the G(i2) GTP-binding protein more efficiently than dimers

Autor: Arcemisbehere, Laure, Sen, Tuhinadri, Boudier, Laure, Balestre, Marie-Noëlle, Gaibelet, Gérald, Detouillon, Emilie, Orcel, Hélène, Mendre, Christiane, Rahmeh, Rita, Granier, Sébastien, Vivès, Corinne, Fieschi, Franck, Damian, Marjorie, Durroux, Thierry, Baneres, Jean-Louis, Mouillac, Bernard
Přispěvatelé: Institut de Génomique Fonctionnelle (IGF), Université de Montpellier (UM)-Université Montpellier 1 (UM1)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Montpellier 2 - Sciences et Techniques (UM2)-Centre National de la Recherche Scientifique (CNRS), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Institut des Biomolécules Max Mousseron [Pôle Chimie Balard] (IBMM), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Montpellier (UM)-Ecole Nationale Supérieure de Chimie de Montpellier (ENSCM), This work was granted by INSERM, CNRS, the European community (#LSHB-CT-2003-503337), the ANRS foundation (#AC14), the French Ministry of Research (ACI BCMS #328) and the National Agency of Research (ANR 06-BLAN-0087). L.A. was the recipient of a PhD fellowship from the French Ligue contre le Cancer. The strategy described here has been patented by INSERM/CNRS (patent 03/07411 and WO 2004/113539 A3), ANR-06-BLAN-0087,GPCR dimers,Molecular and dynamic aspects of G-protein coupled receptor dimers functioning(2006)
Jazyk: angličtina
Rok vydání: 2010
Předmět:
Zdroj: Journal of Biological Chemistry
Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2010, 285 (9), pp.6337-47. ⟨10.1074/jbc.M109.083477⟩
ISSN: 0021-9258
1083-351X
DOI: 10.1074/jbc.M109.083477⟩
Popis: International audience; Accumulating evidence indicates that G protein-coupled receptors can assemble as dimers/oligomers but the role of this phenomenon in G protein coupling and signaling is not yet clear. We have used the purified leukotriene B(4) receptor BLT2 as a model to investigate the capacity of receptor monomers and dimers to activate the adenylyl cyclase inhibitory G(i2) protein. For this, we overexpressed the recombinant receptor as inclusion bodies in the Escherichia coli prokaryotic system, using a human alpha(5) integrin as a fusion partner. This strategy allowed the BLT2 as well as several other G protein-coupled receptors from different families to be produced and purified in large amounts. The BLT2 receptor was then successfully refolded to its native state, as measured by high-affinity LTB(4) binding in the presence of the purified G protein G alpha(i2). The receptor dimer, in which the two protomers displayed a well defined parallel orientation as assessed by fluorescence resonance energy transfer, was then separated from the monomer. Using two methods of receptor-catalyzed guanosine 5'-3-O-(thio)triphosphate binding assay, we clearly demonstrated that monomeric BLT2 stimulates the purified G alpha(i2) beta(1) gamma(2) protein more efficiently than the dimer. These data suggest that assembly of two BLT2 protomers into a dimer results in the reduced ability to signal.
Databáze: OpenAIRE
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