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.

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