Abstract
Fluorescence resonance energy transfer (FRET) is an approach widely used to detect protein–protein interactions in live cells. This approach is based on the sensitization of an “acceptor” molecule by the energy transfer from a “donor” when there is an overlap between the emission spectrum of the “donor” and the excitation spectrum of the “acceptor” and close proximity between the two fluorophore species (in the region of 8 nm). Various methods exist to quantify FRET signals: here, we describe the application of homogeneous time-resolved FRET (htrFRET) combined with Tag-lite™ technology and its application to determine not only protein–protein interactions but also the capability of GPCR mutant variants to form homomers compared to the wild type GPCR within the plasma membrane of transfected cells.
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Acknowledgements
This work was supported by The Medical Research Council (UK) grants [MR/L023806/1 and G0900050] to GM. SM thanks the Istituto Pasteur, Fondazione Cenci-Bolognetti for support. MJVL thanks the Fundación Pedro Barrié de la Maza for support.
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Marsango, S., Varela, M.J., Milligan, G. (2015). Approaches to Characterize and Quantify Oligomerization of GPCRs. 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_7
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DOI: https://doi.org/10.1007/978-1-4939-2914-6_7
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