Metabolites regulate protein function via covalent and noncovalent interactions. However, manipulating these interactions in living cells remains a major challenge. Here, we report a chemical strategy for inducing cysteine S-succination, a nonenzymatic post-translational modification derived from the oncometabolite fumarate. Using a combination of antibody-based detection and kinetic assays, we benchmark the in vitro and cellular reactivity of two novel S-succination "agonists," maleate and 2-bromosuccinate. Cellular assays reveal maleate to be a more potent and less toxic inducer of S-succination, which can activate KEAP1-NRF2 signaling in living cells. By enabling the cellular reconstitution of an oncometabolite-protein interaction with physiochemical accuracy and minimal toxicity, this study provides a methodological basis for better understanding the signaling role of metabolites in disease.

中文翻译：

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Oncometabolite 异构体快速诱导病理生理学蛋白质修饰。

代谢物通过共价和非共价相互作用调节蛋白质功能。然而，在活细胞中操纵这些相互作用仍然是一个重大挑战。在这里，我们报告了一种诱导半胱氨酸 S-琥珀化的化学策略，这是一种源自癌代谢物富马酸盐的非酶促翻译后修饰。结合基于抗体的检测和动力学分析，我们对两种新型 S-琥珀化“激动剂”马来酸盐和 2-溴琥珀酸盐的体外和细胞反应性进行了基准测试。细胞分析表明马来酸盐是一种更有效、毒性更小的 S-琥珀化诱导剂，它可以激活活细胞中的 KEAP1-NRF2 信号传导。通过使癌代谢物-蛋白质相互作用的细胞重建具有理化准确性和最小的毒性，