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.

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.

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

• Journal Article Type: Article
• Acceptance Date: Apr 4, 2015
• Online Publication Date: Jun 1, 2015
• Deposit Date: Jul 19, 2016
• Publicly Available Date: Jul 19, 2016
• Journal: Nature Methods
• Print ISSN: 1548-7091
• Electronic ISSN: 1548-7105
• Publisher: Nature Publishing Group
• Peer Reviewed: Peer Reviewed
• Volume: 12
• DOI: https://doi.org/10.1038/nmeth.3398
• Keywords: cell signalling, drug development, optical spectroscopy
• Public URL: https://nottingham-repository.worktribe.com/output/750676
• Publisher URL: http://www.nature.com/nmeth/journal/v12/n7/full/nmeth.3398.html