Department of Chemistry, Drexel University, Philadelphia, Pennsylvania, USA.
Amgen Inc., Thousand Oaks, California, USA.
J Biol Chem. 2024 Oct;300(10):107784. doi: 10.1016/j.jbc.2024.107784. Epub 2024 Sep 18.
Redox signaling is a fundamental mechanism that controls all major biological processes partly via protein cysteine oxidations, including S-glutathionylation. Despite over 2000 cysteines identified to form S-glutathionylation in databases, the identification of redox cysteines functionally linked to a biological process of interest remains challenging. Here, we demonstrate a strategy combining glutathionylation proteomic database, bioinformatics, and biological screening, which resulted in the identification of S-glutathionylated proteins, including PP2Cα, as redox players of cell migration. We showed that PP2Cα, a prototypical magnesium-dependent serine/threonine phosphatase, is susceptible to S-glutathionylation selectively at nonconserved C314. PP2Cα glutathionylation causes increased migration and invasion of breast cancer cell lines in oxidative stress or upon hydrogen peroxide production. Mechanistically, PP2Cα glutathionylation modulates its protein-protein interactions, activating c-Jun N-terminal kinase and extracellular signal-regulated kinase pathways to elevate migration and invasion. In addition, PP2Cα glutathionylation occurs in response to epidermal growth factor, supporting a serine/threonine phosphatase PP2Cα as a new redox player in growth factor signal transduction.
氧化还原信号是一种基本机制，通过蛋白质半胱氨酸氧化部分控制所有主要的生物过程，包括 S-谷胱甘肽化。尽管在数据库中已经鉴定出超过 2000 个形成 S-谷胱甘肽化的半胱氨酸，但鉴定与感兴趣的生物学过程功能相关的氧化还原半胱氨酸仍然具有挑战性。在这里，我们展示了一种结合谷胱甘肽化蛋白质组数据库、生物信息学和生物筛选的策略，该策略导致了 S-谷胱甘肽化蛋白的鉴定，包括 PP2Cα，作为细胞迁移的氧化还原参与者。我们表明，PP2Cα，一种典型的依赖镁的丝氨酸/苏氨酸磷酸酶，易受 S-谷胱甘肽化，选择性地在非保守的 C314 上。PP2Cα 谷胱甘肽化导致乳腺癌细胞系在氧化应激或产生过氧化氢时迁移和侵袭增加。在机制上，PP2Cα 谷胱甘肽化调节其蛋白质-蛋白质相互作用，激活 c-Jun N 端激酶和细胞外信号调节激酶途径，从而提高迁移和侵袭。此外，PP2Cα 谷胱甘肽化响应表皮生长因子发生，支持丝氨酸/苏氨酸磷酸酶 PP2Cα 作为生长因子信号转导中的新氧化还原参与者。
