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Application of BRET to monitor ligand binding to GPCRs

Stoddart, Leigh A.; Johnstone, Elizabeth K.M.; Wheal, Amanda J.; Goulding, Joelle; Robers, Matthew B.; Machleidt, Thomas; Wood, Keith V.; Hill, Stephen J.; Pfleger, Kevin D.G.

Authors

Elizabeth K.M. Johnstone

Amanda J. Wheal

Matthew B. Robers

Thomas Machleidt

Keith V. Wood

STEPHEN HILL steve.hill@nottingham.ac.uk
Professor of Molecular Pharmacology

Kevin D.G. Pfleger



Abstract

Bioluminescence resonance energy transfer (BRET) is a well-established method for investigating protein-protein interactions. Here we present a BRET approach to monitor ligand binding to G protein–coupled receptors (GPCRs) on the surface of living cells made possible by the use of fluorescent ligands in combination with a bioluminescent protein (NanoLuc) that can be readily expressed on the N terminus of GPCRs.

Journal Article Type Article
Journal Nature Methods
Print ISSN 1548-7091
Electronic ISSN 1548-7105
Publisher Nature Publishing Group
Peer Reviewed Peer Reviewed
Volume 12
APA6 Citation Stoddart, L. A., Johnstone, E. K., Wheal, A. J., Goulding, J., Robers, M. B., Machleidt, T., …Pfleger, K. D. (in press). Application of BRET to monitor ligand binding to GPCRs. Nature Methods, 12, https://doi.org/10.1038/nmeth.3398
DOI https://doi.org/10.1038/nmeth.3398
Keywords cell signalling, drug development, optical spectroscopy
Publisher URL http://www.nature.com/nmeth/journal/v12/n7/full/nmeth.3398.html
Copyright Statement Copyright information regarding this work can be found at the following address: http://eprints.nottingh.../end_user_agreement.pdf

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Copyright Statement
Copyright information regarding this work can be found at the following address: http://eprints.nottingham.ac.uk/end_user_agreement.pdf





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