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.
CITATION STYLE
Masuho, I., Martemyanov, K. A., & Lambert, N. A. (2015). Monitoring G protein activation in cells with BRET. Methods in Molecular Biology, 1335, 107–113. https://doi.org/10.1007/978-1-4939-2914-6_8
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