Bimolecular fluorescence complementation analysis of G protein-coupled receptor dimerization in living cells

Methods Enzymol. 2013;521:259-79. doi: 10.1016/B978-0-12-391862-8.00014-4.

Abstract

Emerging evidence indicates that G protein-coupled receptor (GPCR) signaling is mediated by receptor-receptor interactions at multiple levels. Thus, understanding the biochemistry and pharmacology of those receptor complexes is an important part of delineating the fundamental processes associated with GPCR-mediated signaling in human disease. A variety of experimental approaches have been used to explore these complexes, including bimolecular fluorescence complementation (BiFC) and multicolor BiFC (mBiFC). BiFC approaches have recently been used to explore the composition, cellular localization, and drug modulation of GPCR complexes. The basic methods for applying BiFC and mBiFC to study GPCRs in living cells are the subject of the present chapter.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Fluorescence
  • Fluorometry / methods
  • Humans
  • Microscopy, Fluorescence / methods
  • Protein Interaction Mapping / methods*
  • Protein Multimerization
  • Receptors, G-Protein-Coupled / analysis*
  • Receptors, G-Protein-Coupled / genetics
  • Receptors, G-Protein-Coupled / metabolism*
  • Recombinant Fusion Proteins / analysis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Transfection / methods

Substances

  • Receptors, G-Protein-Coupled
  • Recombinant Fusion Proteins