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.
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Masuho, I., Martemyanov, K.A., Lambert, N.A. (2015). Monitoring G Protein Activation in Cells with BRET. In: Filizola, M. (eds) G Protein-Coupled Receptors in Drug Discovery. Methods in Molecular Biology, vol 1335. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2914-6_8
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DOI: https://doi.org/10.1007/978-1-4939-2914-6_8
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2913-9
Online ISBN: 978-1-4939-2914-6
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