Investigation of Receptor Heteromers Using NanoBRET Ligand Binding

Johnstone, Elizabeth K. M.; See, Heng B.; Abhayawardana, Rekhati S.; Song, Angela; Rosengren, K. Johan; Hill, Stephen J.; Pfleger, Kevin D. G.

Abstract

Receptor heteromerization is the formation of a complex involving at least two different receptors with pharmacology that is distinct from that exhibited by its constituent receptor units. Detection of these complexes and monitoring their pharmacology is crucial for understanding how receptors function. The Receptor-Heteromer Investigation Technology (Receptor-HIT) utilizes liganddependent modulation of interactions between receptors and specific biomolecules for the detection and profiling of heteromer complexes. Previously, the interacting biomolecules used in ReceptorHIT assays have been intracellular proteins, however in this study we have for the first time used bioluminescence resonance energy transfer (BRET) with fluorescently-labeled ligands to investigate heteromerization of receptors on the cell surface. Using the Receptor-HIT ligand binding assay with NanoBRET, we have successfully investigated heteromers between the angiotensin II type 1 (AT1 ) receptor and the β2 adrenergic receptor (AT1-β2AR heteromer), as well as between the AT1 and angiotensin II type 2 receptor (AT1-AT2 heteromer).

Citation

Johnstone, E. K. M., See, H. B., Abhayawardana, R. S., Song, A., Rosengren, K. J., Hill, S. J., & Pfleger, K. D. G. (2021). Investigation of Receptor Heteromers Using NanoBRET Ligand Binding. International Journal of Molecular Sciences, 22(3), Article 1082. https://doi.org/10.3390/ijms22031082

• Journal Article Type: Article
• Acceptance Date: Jan 19, 2021
• Online Publication Date: Jan 22, 2021
• Publication Date: Jan 22, 2021
• Deposit Date: Oct 23, 2023
• Publicly Available Date: Oct 31, 2023
• Journal: International Journal of Molecular Sciences
• Print ISSN: 1661-6596
• Electronic ISSN: 1422-0067
• Publisher: MDPI
• Peer Reviewed: Peer Reviewed
• Volume: 22
• Issue: 3
• Article Number: 1082
• DOI: https://doi.org/10.3390/ijms22031082
• Keywords: Physical and Theoretical Chemistry; Inorganic Chemistry; Organic Chemistry; Spectroscopy; Molecular Biology; Catalysis; General Medicine; Computer Science Applications
• Public URL: https://nottingham-repository.worktribe.com/output/5262443
• Publisher URL: https://www.mdpi.com/1422-0067/22/3/1082