LBPO.CH01 · 化学 · Late-Breaking
化学蛋白质组学发现变构共价GSTO1抑制剂揭示一种此前未被认识、具有抗炎活性的结合模式
Chemoproteomic discovery of allosteric covalent GSTO1 inhibitors reveals a previously unrecognized binding mode with anti-inflammatory activity
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
谷胱甘肽S-转移酶omega 1（GSTO1）通过对NEK7的去谷胱甘肽化促进NLRP3炎症小体的激活，从而促成与癌症和炎症性疾病相关的促炎信号，然而选择性且具细胞活性的GSTO1抑制剂仍然有限。利用一种活细胞PRM-ABPP平台（ChomiXScreenTM），我们鉴定出一种共价小分子抑制剂（A1），其在完整细胞中于Cys32处结合GSTO1，随后通过基于配体的优化生成了一个聚焦的类似物系列。其中，A13表现出持续的GSTO1结合（kinact/KI = 226 M⁻¹·s⁻¹）、经竞争性Ucov-ABPP评估的高全蛋白质组半胱氨酸选择性，以及在人肝微粒体中改善的代谢稳定性。高分辨率晶体结构揭示，A13采取一种出乎意料的变构共价结合模式，不同于已报道的现有GSTO1抑制剂。在功能上，对GSTO1 Cys32的共价靶向显著抑制了原代人单核细胞来源巨噬细胞中LPS诱导的IL-1beta和IL-18分泌。总之，这些发现鉴定出一种此前未被认识的GSTO1抑制变构共价结合模式，并证明了活细胞化学蛋白质组学在发现选择性、功能性共价抑制剂以调节与癌症相关的炎症方面的实用性。
查看英文原文 English abstract
Glutathione S-transferase omega 1 (GSTO1) promotes activation of the NLRP3 inflammasome through deglutathionylation of NEK7, contributing to pro-inflammatory signaling relevant to cancer and inflammatory disease, yet selective and cell-active GSTO1 inhibitors remain limited. Using a live-cell PRM-ABPP platform (ChomiXScreenTM), we identified a covalent small-molecule inhibitor (A1) that engages GSTO1 at Cys32 in intact cells, followed by ligand-based optimization to generate a focused analogue series. Among these, A13 exhibited sustained GSTO1 engagement (kinact/KI = 226 M⁻¹·s⁻¹), high proteome-wide cysteine selectivity as assessed by competitive Ucov-ABPP, and improved metabolic stability in human liver microsomes. High-resolution crystal structures revealed that A13 adopts an unexpected allosteric covalent binding mode distinct from those reported for existing GSTO1 inhibitors. Functionally, covalent targeting of GSTO1 Cys32 significantly suppressed LPS-induced IL-1beta and IL-18 secretion in primary human monocyte-derived macrophages. Together, these findings identify a previously unrecognized allosteric covalent binding mode for GSTO1 inhibition and demonstrate the utility of live-cell chemoproteomics for discovering selective, functional covalent inhibitors to modulate inflammation relevant to cancer.
利益披露 Disclosure
N. Chen, None..
F. Zhang, None..
W. Lu, None.