Biochemistry & Molecular Biology Program, Departments of Biology and Chemistry, The College of Wooster, Wooster, Ohio 44691, United States.
Chem Res Toxicol. 2022 Oct 17;35(10):1676-1689. doi: 10.1021/acs.chemrestox.2c00123. Epub 2022 Jun 30.
Reversible oxidation of cysteine residues within proteins occurs naturally during normal cellular homeostasis and can increase during oxidative stress. Cysteine oxidation often leads to the formation of disulfide bonds, which can impact protein folding, stability, and function. Work in both prokaryotic and eukaryotic models over the past five decades has revealed several multiprotein systems that use thiol-dependent oxidoreductases to mediate disulfide bond reduction, formation, and/or rearrangement. Here, I provide an overview of how these systems operate to carry out disulfide exchange reactions in different cellular compartments, with a focus on their roles in maintaining redox homeostasis, transducing redox signals, and facilitating protein folding. Additionally, I review thiol-independent and thiol-dependent approaches for interrogating what proteins partner together in such disulfide-based redox relays. While the thiol-independent approaches rely either on predictive measures or standard procedures for monitoring protein-protein interactions, the thiol-dependent approaches include direct disulfide trapping methods as well as thiol-dependent chemical cross-linking. These strategies may prove useful in the systematic characterization of known and newly discovered disulfide relay mechanisms and redox switches involved in oxidant defense, protein folding, and cell signaling.
蛋白质中半胱氨酸残基的氧化还原是蛋白质翻译后修饰的一种重要类型，在正常细胞内环境中自然发生，并可在氧化应激时增加。半胱氨酸氧化通常导致二硫键的形成，这会影响蛋白质的折叠、稳定性和功能。过去五十年中，在原核和真核模型中的研究揭示了几种利用硫醇依赖性氧化还原酶来介导二硫键还原、形成和/或重排的多蛋白系统。在这里，我概述了这些系统如何在不同的细胞区室中进行二硫键交换反应，重点介绍它们在维持氧化还原稳态、传递氧化还原信号和促进蛋白质折叠方面的作用。此外，我还回顾了用于研究这些二硫键氧化还原中继中哪些蛋白质相互作用的硫醇独立和硫醇依赖方法。虽然硫醇独立的方法依赖于预测性措施或监测蛋白质-蛋白质相互作用的标准程序，但硫醇依赖的方法包括直接二硫键捕获方法以及硫醇依赖性化学交联。这些策略可能有助于系统地表征已知和新发现的参与抗氧化防御、蛋白质折叠和细胞信号转导的二硫键中继机制和氧化还原开关。
