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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., Vandenabeele, S., Zabel, U., Frank, M., 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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