Abstract
The spatiotemporal characterization of protein–protein interactions is essential to understand nearly all cellular events. Several methodological strategies derived from noninvasive luminescence- and fluorescence-based approaches allow the detection of specific protein–protein interactions in living cells. Here, we describe the application of bioluminescence resonance energy transfer (BRET) and donor fluorescent recovery after photobleaching (dFRAP) approaches to the study of G protein-coupled receptor (GPCR) oligomerization. These technologies alone – or in combination with complementary methods – can become extremely powerful approaches for visualizing these cellular protein–protein interactions, even between more than two proteins. Overall, these methods might become extremely important tools in GPCR drug discovery.
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Acknowledgments
This work was supported by grants SAF2011-24779, Consolider-Ingenio CSD2008-00005, and PCIN-2013-019-C03-03 from Ministerio de Economía y Competitividad and ICREA Academia-2010 from the Catalan Institution for Research and Advanced Studies to F.C. Also, the authors belong to the “Neuropharmacology and Pain” accredited research group (Generalitat de Catalunya, 2014 SGR 1251).
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Ciruela, F., Fernández-Dueñas, V. (2015). GPCR Oligomerization Analysis by Means of BRET and dFRAP. In: Prazeres, D.M.F., Martins, S.A.M. (eds) G Protein-Coupled Receptor Screening Assays. Methods in Molecular Biology, vol 1272. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-2336-6_10
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DOI: https://doi.org/10.1007/978-1-4939-2336-6_10
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