Itaconate is an anti-inflammatory metabolite involved in pathogen-macrophage interaction, but the mechanisms underlying its effect are not fully understood. Competitive cysteine profiling has been performed to interrogate itaconate's reactivity in cell lysates, however, methods for analyzing targets of itaconation directly in living macrophages are still lacking. Here we developed a specific bioorthogonal probe, itaconate-alkyne (ITalk), for quantitative and site-specific chemoproteomic profiling of itaconation in inflammatory macrophages. ITalk recapitulates the anti-inflammatory property of itaconate and enables biochemical evaluation and proteomic analysis of its direct targets. Our study delineates the widespread landscape of itaconate substrates, providing a versatile tool and comprehensive resource for investigating its function.

中文翻译：

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炎症巨噬细胞中生物正交探针对衣康宁的化学蛋白质组学分析

衣康酸盐是一种参与病原体-巨噬细胞相互作用的抗炎代谢物，但其作用机制尚不完全清楚。已经进行了竞争性半胱氨酸分析以询问细胞裂解物中衣康酸的反应性，但是，仍然缺乏直接在活巨噬细胞中分析衣康酸靶标的方法。在这里，我们开发了一种特定的生物正交探针，衣康酸酯-炔烃 (ITalk)，用于炎症性巨噬细胞中衣康酸酯的定量和位点特异性化学蛋白质组学分析。ITalk 概括了衣康酸酯的抗炎特性，并能够对其直接目标进行生化评估和蛋白质组学分析。我们的研究描绘了衣康酸酯底物的广泛景观，为研究其功能提供了通用工具和综合资源。