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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

DOI: 10.1021/acsbiomedchemau.3c00060 DOI Help

Authors: Alexander J. Lander (Cardiff University) , Yifu Kong (Xiamen University) , Yi Jin (University of Manchester) , Chuanliu Wu (Xiamen University) , Louis Y. P. Luk (Cardiff University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acs Bio & Med Chem Au

State: Published (Approved)
Published: December 2023
Diamond Proposal Number(s): 20147

Open Access Open Access

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.

Journal Keywords: Chemical protein synthesis; protein folding; cysteine-rich protein; mirror-image phage display; racemic protein crystallography; d-protein; d-peptide

Subject Areas: Biology and Bio-materials, Chemistry, Medicine

Instruments: I03-Macromolecular Crystallography

Added On: 13/12/2023 10:48

Discipline Tags:

Health & Wellbeing Biochemistry Chemistry Structural biology Biophysics Drug Discovery Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)