Analysis of the coupling of G12/13 to G protein-coupled receptors using a luciferase reporter assay.
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.
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000378:metabolism*; D017930:Genes, Reporter; D008156:Luciferases / Q000235:genetics*; D051379:Mice; D041681:NIH 3T3 Cells; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection / Q000379:methods*
D043562:Receptors, G-Protein-Coupled; D008156:Luciferases; D044365:GTP-Binding Protein alpha Subunits, G12-G13