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., Wood, K. V., Hill, S. J., & 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
• Contract Date: Jul 19, 2016