Deciphering the synthetic and refolding strategy of a cysteine-rich domain in the tumor necrosis factor receptor (TNF-R) for racemic crystallography analysis and d-peptide ligand discovery

Lander, Alexander J., Kong, Yifu, Jin, Yi, Wu, Chuanliu and Luk, Louis Y. P. ORCID: https://orcid.org/0000-0002-7864-6261 2024. Deciphering the synthetic and refolding strategy of a cysteine-rich domain in the tumor necrosis factor receptor (TNF-R) for racemic crystallography analysis and d-peptide ligand discovery. ACS Bio & Med Chem Au 4 (1) , pp. 68-76. 10.1021/acsbiomedchemau.3c00060

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Abstract

Many cell-surface receptors are promising targets for chemical synthesis because of their critical roles in disease development. This synthetic approach enables investigations by racemic protein crystallography and ligand discovery by mirror-image methodologies. However, due to their complex nature, the chemical synthesis of a receptor can be a significant challenge. Here, we describe the chemical synthesis and folding of a central, cysteine-rich domain of the cell-surface receptor tumor necrosis factor 1 which is integral to binding of the cytokine TNF-α, namely, TNFR-1 CRD2. Racemic protein crystallography at 1.4 Å confirmed that the native binding conformation was preserved, and TNFR-1 CRD2 maintained its capacity to bind to TNF-α (KD ≈ 7 nM). Encouraged by this discovery, we carried out mirror-image phage display using the enantiomeric receptor mimic and identified a d-peptide ligand for TNFR-1 CRD2 (KD = 1 μM). This work demonstrated that cysteine-rich domains, including the central domains, can be chemically synthesized and used as mimics for investigations.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Chemistry
Publisher:	American Chemical Society
ISSN:	2694-2437
Funders:	BBSRC, Wellcome Trust
Date of First Compliant Deposit:	14 December 2023
Date of Acceptance:	15 November 2023
Last Modified:	12 Aug 2024 09:26
URI:	https://orca.cardiff.ac.uk/id/eprint/164809

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