Cell-surface protein-protein interaction analysis with time-resolved FRET and snap-tag technologies: Application to GPCR oligomerization

Damien Maurel, Laëtitia Comps-Agrar, Carsten Brock, Marie Laure Rives, Emmanuel Bourrier, Mohammed Akli Ayoub, Hervé Bazin, Norbert Tinel, Thierry Durroux, Laurent Prézeau, Eric Trinquet, Jean Philippe Pin

Research output: Contribution to journalArticlepeer-review

405 Citations (Scopus)

Abstract

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 γ-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.

Original languageEnglish
Pages (from-to)561-567
Number of pages7
JournalNature Methods
Volume5
Issue number6
DOIs
Publication statusPublished - Jun 2008
Externally publishedYes

ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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