Skip to main content

Research Repository

Advanced Search

Probing the binding of interleukin-23 to individual receptor components and the IL-23 heteromeric receptor complex in living cells using NanoBRET

Lay, Charles S; Bridges, Angela; Goulding, Joelle; Briddon, Stephen J; Soloviev, Zoja; Craggs, Peter D; Hill, Stephen J.

Probing the binding of interleukin-23 to individual receptor components and the IL-23 heteromeric receptor complex in living cells using NanoBRET Thumbnail


Authors

Charles S Lay

Angela Bridges

Zoja Soloviev

Peter D Craggs

STEPHEN HILL STEVE.HILL@NOTTINGHAM.AC.UK
Professor of Molecular Pharmacology



Abstract

Interleukin-23 (IL-23) is a pro-inflammatory cytokine involved in the host defence against pathogens, but also implicated in the development of several autoimmune disorders. The IL- 23 receptor has become a key target for drug discovery but the exact mechanism of the receptor ligand interaction remains poorly understood. In this study the affinities of IL-23 for its individual receptor components (IL23R and IL12Rβ1) and the heteromeric complex formed between them have been measured in living cells using NanoLuciferase tagged full-length proteins. Here, we demonstrate that TAMRA tagged IL-23 has a greater than seven fold higher affinity for IL12Rβ1 than IL23R. However, in the presence of both receptor subunits IL-23 affinity is increased more than three orders of magnitude to 27 pM. Furthermore, we show that IL-23 induces a potent change in the position of the N-terminal domains of the two receptor subunits consistent with a conformational change in the heteromeric receptor structure.

Citation

Lay, C. S., Bridges, A., Goulding, J., Briddon, S. J., Soloviev, Z., Craggs, P. D., & Hill, S. J. (2022). Probing the binding of interleukin-23 to individual receptor components and the IL-23 heteromeric receptor complex in living cells using NanoBRET. Cell Chemical Biology, 29(1), 19-29.e6. https://doi.org/10.1016/j.chembiol.2021.05.002

Journal Article Type Article
Acceptance Date May 4, 2021
Online Publication Date May 25, 2021
Publication Date Jan 20, 2022
Deposit Date May 12, 2021
Publicly Available Date May 26, 2022
Journal Cell Chemical Biology
Publisher Elsevier
Peer Reviewed Peer Reviewed
Volume 29
Issue 1
Pages 19-29.e6
DOI https://doi.org/10.1016/j.chembiol.2021.05.002
Keywords Clinical Biochemistry; Molecular Medicine; Biochemistry; Molecular Biology; Pharmacology; Drug Discovery
Public URL https://nottingham-repository.worktribe.com/output/5527070
Publisher URL https://www.sciencedirect.com/science/article/pii/S2451945621002191

Files





You might also like



Downloadable Citations