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

Schihada, Hannes; Vandenabeele, Sylvie; Zabel, Ulrike; Frank, Monika; Lohse, Martin J.; Maiellaro, Isabella

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


Authors

Hannes Schihada

Sylvie Vandenabeele

Ulrike Zabel

Monika Frank

Martin J. Lohse



Contributors

Hannes Schihada
Data Collector

Sylvie Vandenabeele
Data Collector

Ulrike Zabel
Other

Monika Frank
Other

Martin J. Lohse
Project Leader

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 ?2A-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 ?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.

Citation

Schihada, H., Vandenabeele, S., Zabel, U., Frank, M., Lohse, M. J., & Maiellaro, I. (2018). A universal bioluminescence resonance energy transfer sensor design enables high-sensitivity screening of GPCR activation dynamics. Communications Biology, 1, Article 105. https://doi.org/10.1038/s42003-018-0072-0

Journal Article Type Article
Acceptance Date May 11, 2018
Online Publication Date Aug 7, 2018
Publication Date Dec 1, 2018
Deposit Date Apr 1, 2020
Publicly Available Date Mar 28, 2024
Journal Communications Biology
Electronic ISSN 2399-3642
Publisher Nature Publishing Group
Peer Reviewed Peer Reviewed
Volume 1
Article Number 105
DOI https://doi.org/10.1038/s42003-018-0072-0
Public URL https://nottingham-repository.worktribe.com/output/4230650
Publisher URL https://www.nature.com/articles/s42003-018-0072-0
Additional Information Received: 28 November 2017; Accepted: 11 May 2018; First Online: 7 August 2018; : The authors declare that The University of Würzburg holds a patent on this technology: WO2004057333 A1.

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