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

IF 3.8 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY ACS Bio & Med Chem Au Pub Date : 2023-12-11 DOI:10.1021/acsbiomedchemau.3c00060
Alexander J. Lander, Yifu Kong, Yi Jin*, Chuanliu Wu* and Louis Y. P. Luk*, 
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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.

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破译肿瘤坏死因子受体(TNF-R)富半胱氨酸结构域的合成和重折叠策略,以进行消旋结晶学分析和多肽配体的发现
由于许多细胞表面受体在疾病发展中起着至关重要的作用,因此是化学合成的理想靶标。这种合成方法可以通过外消旋蛋白质晶体学进行研究,并通过镜像方法发现配体。然而,由于受体的复杂性,受体的化学合成可能是一个巨大的挑战。在这里,我们描述了细胞表面受体肿瘤坏死因子 1 的一个富含半胱氨酸的中心结构域(即 TNFR-1 CRD2)的化学合成和折叠,该结构域与细胞因子 TNF-α 的结合密不可分。1.4 Å 的消旋蛋白晶体学证实,原生结合构象得以保留,TNFR-1 CRD2 保持了与 TNF-α 结合的能力(KD ≈ 7 nM)。受到这一发现的鼓舞,我们利用对映体受体模拟物进行了镜像噬菌体展示,并确定了 TNFR-1 CRD2 的 d 肽配体(KD = 1 μM)。这项工作表明,富含半胱氨酸的结构域(包括中心结构域)可以通过化学合成作为模拟物进行研究。
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来源期刊
ACS Bio & Med Chem Au
ACS Bio & Med Chem Au 药物、生物、化学-
CiteScore
4.10
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0.00%
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0
期刊介绍: ACS Bio & Med Chem Au is a broad scope open access journal which publishes short letters comprehensive articles reviews and perspectives in all aspects of biological and medicinal chemistry. Studies providing fundamental insights or describing novel syntheses as well as clinical or other applications-based work are welcomed.This broad scope includes experimental and theoretical studies on the chemical physical mechanistic and/or structural basis of biological or cell function in all domains of life. It encompasses the fields of chemical biology synthetic biology disease biology cell biology agriculture and food natural products research nucleic acid biology neuroscience structural biology and biophysics.The journal publishes studies that pertain to a broad range of medicinal chemistry including compound design and optimization biological evaluation molecular mechanistic understanding of drug delivery and drug delivery systems imaging agents and pharmacology and translational science of both small and large bioactive molecules. Novel computational cheminformatics and structural studies for the identification (or structure-activity relationship analysis) of bioactive molecules ligands and their targets are also welcome. The journal will consider computational studies applying established computational methods but only in combination with novel and original experimental data (e.g. in cases where new compounds have been designed and tested).Also included in the scope of the journal are articles relating to infectious diseases research on pathogens host-pathogen interactions therapeutics diagnostics vaccines drug-delivery systems and other biomedical technology development pertaining to infectious diseases.
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