Analysis of the coupling of G12/13 to G protein-coupled receptors using a luciferase reporter assay

Methods Mol Biol. 2004:237:145-9. doi: 10.1385/1-59259-430-1:145.

Abstract

G protein-coupled receptors (GPCRs) transduce signals from cell surface to intracellular effectors through various G proteins. In this chapter, a luciferase reporter gene assay system is described for the study of the coupling of GPCRs to G alpha 12/13. This assay system, in which ligand-stimulated production of the reporter luciferase under the control of a modified serum-responsive element (SRE) in a cell line derived from mice lacking G alpha q/11 that are transfected with a GPCR of interest, allows for easy, fast, and valid determination of the coupling of the GPCR to G alpha 12/13.

Publication types

  • Review

MeSH terms

  • Animals
  • Cell Culture Techniques / methods
  • GTP-Binding Protein alpha Subunits, G12-G13 / metabolism*
  • Genes, Reporter
  • Luciferases / genetics*
  • Mice
  • NIH 3T3 Cells
  • Receptors, G-Protein-Coupled / metabolism*
  • Signal Transduction
  • Transfection / methods*

Substances

  • Receptors, G-Protein-Coupled
  • Luciferases
  • GTP-Binding Protein alpha Subunits, G12-G13