Visualising ligand-binding to a GPCR in vivo using nanoBRET

Carvalheira Alcobia, Diana; Ziegler, Alexandra I.; Kondrashov, Alexander; Comeo, Eleonora; Mistry, Sarah; Kellam, Barrie; Chang, Aeson; Woolard, Jeannette; Hill, Stephen J.; Sloan, Erica K.

Abstract

© 2018 The Author(s) The therapeutic action of a drug depends on its ability to engage with its molecular target in vivo. However, current drug discovery strategies quantify drug levels within organs rather than determining the binding of drugs directly to their specific molecular targets in vivo. This is a particular problem for assessing the therapeutic potential of drugs that target malignant tumors where access and binding may be impaired by disrupted vasculature and local hypoxia. Here we have used triple-negative human breast cancer cells expressing β2-adrenoceptors tagged with the bioluminescence protein NanoLuc to provide a bioluminescence resonance energy transfer approach to directly quantify ligand binding to a G protein-coupled receptor in vivo using a mouse model of breast cancer.

Citation

Carvalheira Alcobia, D., Ziegler, A. I., Kondrashov, A., Comeo, E., Mistry, S., Kellam, B., Chang, A., Woolard, J., Hill, S. J., & Sloan, E. K. (2018). Visualising ligand-binding to a GPCR in vivo using nanoBRET. iScience, 6(8), 280-288. https://doi.org/10.1016/j.isci.2018.08.006

• Journal Article Type: Article
• Acceptance Date: Aug 6, 2018
• Online Publication Date: Aug 11, 2018
• Publication Date: Aug 31, 2018
• Deposit Date: Aug 9, 2018
• Publicly Available Date: Oct 12, 2018
• Journal: iScience
• Electronic ISSN: 2589-0042
• Publisher: Cell Press
• Peer Reviewed: Peer Reviewed
• Volume: 6
• Issue: 8
• Pages: 280-288
• DOI: https://doi.org/10.1016/j.isci.2018.08.006
• Public URL: https://nottingham-repository.worktribe.com/output/986075
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S2589004218301184
• Additional Information: This article is maintained by: Elsevier; Article Title: Visualizing Ligand Binding to a GPCR InVivo Using NanoBRET; Journal Title: iScience; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.isci.2018.08.006; Content Type: article; Copyright: © 2018 The Author(s).
• Contract Date: Aug 9, 2018