Abu Hariri Hiba, Braunstein Ilana, Salti Talal, Glaser Fabian, Gefen Tal, Geva-Zatorsky Naama, Ziv Tamar, Benhar Moran
Department of Biochemistry, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa, Israel.
Bioinformatic Knowledge Unit, The Lorry I. Lokey Interdisciplinary Center for Life Sciences and Engineering, Technion-Israel Institute of Technology, Haifa, Israel.
Antioxid Redox Signal. 2023 Feb;38(4-6):388-402. doi: 10.1089/ars.2022.0026. Epub 2022 Oct 18.
Oxidative modifications of cysteine (Cys) thiols regulate various physiological processes, including inflammatory responses. The thioredoxin (Trx) system plays a key role in thiol redox control. The aim of this study was to characterize the dynamic cysteine proteome of human macrophages upon activation by the prototypical proinflammatory agent, bacterial lipopolysaccharide (LPS), and/or perturbation of the Trx system. In this study, we profiled the cellular and redox proteome of human THP-1-derived macrophages during the early phase of LPS activation and/or inhibition of Trx system activity by auranofin (AF) by employing a peptide-centric, resin-assisted capture, redox proteomic workflow. Among 4200 identified cysteines, oxidation of nearly 10% was selectively affected by LPS or AF treatments. Notably, the proteomic analysis uncovered a subset of ∼100 thiols, mapped to proteins involved in diverse processes, whose oxidation is antagonistically regulated by LPS and Trx. Compared with the redox proteome, the cellular proteome was largely unchanged, highlighting the importance of redox modification as a mechanism that allows for rapid modulation of macrophage activities in response to a proinflammatory or pro-oxidant insult. Structural-functional analyses provided mechanistic insights into redox regulation of selected proteins, including the glutathione-synthesizing enzyme, glutamate-cysteine ligase, and the autophagy adaptor, SQSTM1/p62, suggesting mechanisms by which macrophages adapt and fine-tune their responses according to a changing inflammatory and redox environment. This study provides a rich resource for further characterization of redox mechanisms that regulate macrophage inflammatory activities. The dynamic thiol redox proteome allows macrophages to efficiently respond and adapt to redox and inflammatory challenges. 38, 388-402.
半胱氨酸（Cys）硫醇的氧化修饰调节多种生理过程，包括炎症反应。硫氧还蛋白（Trx）系统在硫醇氧化还原控制中起关键作用。本研究的目的是表征人类巨噬细胞在受到典型促炎剂细菌脂多糖（LPS）激活和/或硫氧还蛋白系统受到干扰时的动态半胱氨酸蛋白质组。在本研究中，我们采用以肽为中心、树脂辅助捕获的氧化还原蛋白质组学工作流程，分析了人THP-1来源的巨噬细胞在LPS激活早期阶段和/或金诺芬（AF）抑制硫氧还蛋白系统活性过程中的细胞蛋白质组和氧化还原蛋白质组。在鉴定出的4200个半胱氨酸中，近10%的氧化受到LPS或AF处理的选择性影响。值得注意的是，蛋白质组分析发现了大约100个硫醇的子集，这些硫醇映射到参与不同过程的蛋白质上，其氧化受到LPS和硫氧还蛋白的拮抗调节。与氧化还原蛋白质组相比，细胞蛋白质组基本未发生变化，这突出了氧化还原修饰作为一种机制的重要性，该机制能够使巨噬细胞活动快速响应促炎或促氧化刺激而进行调节。结构功能分析为选定蛋白质的氧化还原调节提供了机制性见解，包括谷胱甘肽合成酶、谷氨酸-半胱氨酸连接酶和自噬衔接蛋白SQSTM1/p62，这表明巨噬细胞根据不断变化的炎症和氧化还原环境进行适应和微调反应的机制。本研究为进一步表征调节巨噬细胞炎症活动的氧化还原机制提供了丰富资源。动态硫醇氧化还原蛋白质组使巨噬细胞能够有效应对并适应氧化还原和炎症挑战。38, 388 - 402。
