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A universal bioluminescence resonance energy transfer sensor design enables high-sensitivity screening of GPCR activation dynamics

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Item Type:Article
Title:A universal bioluminescence resonance energy transfer sensor design enables high-sensitivity screening of GPCR activation dynamics
Creators Name:Schihada, H. and Vandenabeele, S. and Zabel, U. and Frank, M. and Lohse, M.J. and Maiellaro, I.
Abstract:G-protein-coupled receptors (GPCRs) represent one of the most important classes of drug targets. The discovery of new GCPR therapeutics would greatly benefit from the development of a generalizable high-throughput assay to directly monitor their activation or de-activation. Here we screened a variety of labels inserted into the third intracellular loop and the C-terminus of the α-adrenergic receptor and used fluorescence (FRET) and bioluminescence resonance energy transfer (BRET) to monitor ligand-binding and activation dynamics. We then developed a universal intramolecular BRET receptor sensor design to quantify efficacy and potency of GPCR ligands in intact cells and real time. We demonstrate the transferability of the sensor design by cloning β-adrenergic and PTH1-receptor BRET sensors and monitored their efficacy and potency. For all biosensors, the Z factors were well above 0.5 showing the suitability of such design for microtiter plate assays. This technology will aid the identification of novel types of GPCR ligands.
Source:Communications Biology
ISSN:2399-3642
Volume:1
Page Range:105
Date:7 August 2018
Official Publication:https://doi.org/10.1038/s42003-018-0072-0
PubMed:View item in PubMed

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