title: Monitoring G Protein Activation in Cells with BRET.
abstract: Live-cell assays based on fluorescence and luminescence are now indispensable tools for the study of G protein signaling. Assays based on fluorescence and bioluminescence resonance energy transfer (FRET and BRET) have been particularly valuable for monitoring changes in second messengers, protein-protein interactions, and protein conformation. Here, we describe a BRET assay that monitors the release of free Gβγ dimers after activation of heterotrimers containing Gα subunits from all four G protein subfamilies. This assay provides useful kinetic and pharmacological information with reasonably high throughput using a standard laboratory equipment.
authors.affiliation: Department of Neuroscience, The Scripps Research Institute Florida, Jupiter, FL, 33458, USA.
authors.forename: Ikuo
authors.initials: I
authors.lastname: Masuho
authors.forename: Kirill A
authors.initials: KA
authors.lastname: Martemyanov
authors.forename: Nevin A
authors.initials: NA
authors.lastname: Lambert
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D007202:Indicators and Reagents / Q000737:chemistry; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D013050:Spectrometry, Fluorescence; D014162:Transfection
chemical_list: D007202:Indicators and Reagents; D019204:GTP-Binding Proteins