TY  - JOUR
A1  - Schihada, Hannes
A1  - Vandenabeele, Sylvie
A1  - Zabel, Ulrike
A1  - Frank, Monika
A1  - Lohse, Martin J.
A1  - Maiellaro, Isabella
T1  - A universal bioluminescence resonance energy transfer sensor design enables high-sensitivity screening of GPCR activation dynamics
T2  - Communications Biology
N2  - 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 alpha(2 Lambda)-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 beta(2)-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.
KW  - Fluorescence resonance energy transfer
KW  - G protein-coupled receptors
KW  - High-throughput screening
Y1  - 2018
UR  - https://opus.bibliothek.uni-wuerzburg.de/frontdoor/index/index/docId/22859
UR  - https://nbn-resolving.org/urn:nbn:de:bvb:20-opus-228592
VL  - 1
IS  - 105
ER  -