Nature Methods
- 5, 561 - 567 (2008)
Published online: 18 May 2008; | doi:10.1038/nmeth.1213
Cell-surface protein-protein interaction analysis with time-resolved FRET and snap-tag technologies: application to GPCR oligomerizationDamien Maurel1, 2, 3, 4, 5, Laëtitia Comps-Agrar1, 2, 5, Carsten Brock1, 2, Marie-Laure Rives1, 2, Emmanuel Bourrier3, Mohammed Akli Ayoub1, 2, Hervé Bazin3, Norbert Tinel3, Thierry Durroux1, 2, Laurent Prézeau1, 2, Eric Trinquet3 & Jean-Philippe Pin1, 21
Centre National de la Recherche Scientifique (CNRS), UMR 5203, Institut de Génomique Fonctionnelle, 141 Rue de la Cardonille, Montpellier F-34000, France. 2
Institut National de la Santé et de la Recherche Médicale (INSERM), U661, 141 Rue de la Cardonille, Montpellier F-34000, France and Université Montpellier 1 and Université Montpellier 2, 141 Rue de la Cardonille, Montpellier F-34000, France. 3
Cisbio, Parc technologique Marcel Boiteux, Bagnols/Cèze cedex F-30204, France. 4
Present address: Ecole Polytechnique Fédérale de Lausanne, Institute of Chemical Sciences and Engineering, CH-1015 Lausanne, Switzerland. 5
These authors contributed equally to this work.
Correspondence should be addressed to Jean-Philippe Pin jppin@igf.cnrs.fr Cell-surface proteins are important in cell-cell communication. They assemble into heterocomplexes that include different receptors and effectors. Elucidation and manipulation of such protein complexes offers new therapeutic possibilities. We describe a methodology combining time-resolved fluorescence resonance energy transfer (FRET) with snap-tag technology to quantitatively analyze protein-protein interactions at the surface of living cells, in a high throughput–compatible format. Using this approach, we examined whether G protein–coupled receptors (GPCRs) are monomers or assemble into dimers or larger oligomers—a matter of intense debate. We obtained evidence for the oligomeric state of both class A and class C GPCRs. We also observed different quaternary structure of GPCRs for the neurotransmitters glutamate and  -aminobutyric acid (GABA): whereas metabotropic glutamate receptors assembled into strict dimers, the GABAB receptors spontaneously formed dimers of heterodimers, offering a way to modulate G-protein coupling efficacy. This approach will be useful in systematic analysis of cell-surface protein interaction in living cells.
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