Interaction of protease-activated receptor 2 with G proteins and β-arrestin 1 studied by bioluminescence resonance energy transfer

Mohammed Akli Ayoub, Jean Philippe Pin

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

G protein-coupled receptors are well recognized as being able to activate several signaling pathways through the activation of different G proteins as well as other signaling proteins such as β-arrestins. Therefore, understanding how such multiple GPCR-mediated signaling can be integrated constitute an important aspect. Here, we applied bioluminescence resonance energy transfer (BRET) to shed more light on the G protein coupling profile of trypsin receptor, or protease-activated receptor 2 (PAR2), and its interaction with β-arrestin1. Using YFP and Rluc fusion constructs expressed in COS-7 cells, BRET data revealed a pre-assembly of PAR2 with both Gαi1 and Gαo and a rapid and transient activation of these G proteins upon receptor activation. In contrast, no pre-assembly of PAR2 with Gα12 could be detected and their physical association can be measured with a very slow and sustained kinetics similar to that of β-arrestin1 recruitment. These data demonstrate the coupling of PAR2 with Gαi1, Gαo, and Gα12 in COS-7 cells with differences in the kinetics of GPCR-G protein coupling, a parameter that very likely influences the cellular response. Moreover, this further illustrates that pre-assembly or agonist-induced G protein interaction depends on receptor-G protein pairs indicating another level of complexity and regulation of the signaling of GPCR-G protein complexes and its multiplicity.

Original languageEnglish
Article numberArticle 196
JournalFrontiers in Endocrinology
Volume4
Issue numberDEC
DOIs
Publication statusPublished - 2013
Externally publishedYes

Keywords

  • BRET
  • G proteins
  • PAR2
  • Pre-assembly
  • Trypsin
  • β-arrestin

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism

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