Chapter title |
Monitoring G Protein Activation in Cells with BRET
|
---|---|
Chapter number | 8 |
Book title |
G Protein-Coupled Receptors in Drug Discovery
|
Published in |
Methods in molecular biology, January 2015
|
DOI | 10.1007/978-1-4939-2914-6_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2913-9, 978-1-4939-2914-6
|
Authors |
Ikuo Masuho, Kirill A. Martemyanov, Nevin A. Lambert, Masuho, Ikuo, Martemyanov, Kirill A., Lambert, Nevin A. |
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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Demographic breakdown
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Student > Bachelor | 6 | 12% |
Researcher | 5 | 10% |
Student > Master | 5 | 10% |
Professor | 3 | 6% |
Other | 7 | 14% |
Unknown | 16 | 31% |
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