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Methods Mol Biol. 2015;1335:107-13. doi: 10.1007/978-1-4939-2914-6_8.

Monitoring G Protein Activation in Cells with BRET.

Author information

1
Department of Neuroscience, The Scripps Research Institute Florida, Jupiter, FL, 33458, USA.

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.

PMID:
26260597
PMCID:
PMC4879496
DOI:
10.1007/978-1-4939-2914-6_8
[Indexed for MEDLINE]
Free PMC Article

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