Chemical Tools for Probing Cysteine Sulfenation and Sulfination Redox Biology

项目摘要 半胱氨酸残基的氧化还原状态的翻译后变化可以快速且可逆地改变蛋白质功能， 调节生物过程和药物药理学。在上一个资助期间，我们的实验室报告了几个 有机硫化学、小分子工具和蛋白质组学分析计算方法的创新 这极大地提高了半胱氨酸氧化还原体的选择性、细胞应用和位点特异性定量 (J. Am.化学会，2017; Nat. Chem. Biol.，2018; Nat. Chem.，2021; Nat. Comm.，2022年）。应用该等 化学蛋白质组学方法有助于有意义地发现氧化还原生物学中的新范例（Nat. Cell 生物学：2019;细胞代谢，2019;血液高级，2020年;美国国家通讯社，2021;氧化还原生物学，2021 & 2022; Proc. Natl. Acad. 科学，2022），并提出了关于细胞氧化还原景观，分子 硫醇基氧化还原开关，以及新兴的氧化还原医学领域。以下三个具体目标 提出：1）化学方法的发展和应用，以解决时空控制氧化还原 信号传导; 2）在蛋白质中基因掺入氧化半胱氨酸以查明潜在的分子机制 基于硫醇的氧化还原调节;和3）亲核片段文库，以发现靶向氧化还原的共价配体。 半胱氨酸组中的调节蛋白。强有力的初步数据表明，本文提出的研究都是 在我们的手中是可行的。这些研究成果包括：1）有价值的新化学品 生物学方法来理解硫醇基氧化还原信号的基本机制，和2）新的化学， 这些物质可以作为小分子探针的来源和药物发现的起点。

PROJECT SUMMARY Post-translational changes in the redox state of cysteine residues can rapidly and reversibly alter protein function, modulating biological processes and drug pharmacology. During the last funding period, our lab reported several innovations in organosulfur chemistry, small-molecule tools, and computational methods for proteomic analysis that dramatically improved selectivity, cellular application, and site-specific quantitation of the cysteine redoxome (J. Am. Chem. Soc., 2017; Nat. Chem. Biol., 2018; Nat. Chem., 2021; Nat. Comm., 2022). Application of these chemoproteomic methods has contributed to meaningful discovery of new paradigms in redox biology (Nat. Cell Biol., 2019; Cell Metab., 2019; Blood Adv., 2020; Nat. Comm., 2021; Redox Biol., 2021 & 2022; Proc. Natl. Acad. Sci., 2022) and raise interesting new questions about the links between the cellular redox landscape, molecular thiol-based redox switches, and the emerging field of redox medicine. Herein, the following three Specific Aims are proposed: Development and application of 1) chemical methods to address spatiotemporal control in redox signaling; 2) genetic incorporation of oxidized cysteine in proteins to pinpoint molecular mechanisms underlying thiol-based redox regulation; and 3) nucleophile-fragment libraries to discover covalent ligands that target redox- regulated proteins in the cysteinome. Strong preliminary data demonstrate that studies proposed herein are both promising and feasible in our hands. Deliverables resulting from these studies include: 1) valuable new chemical biology approaches to understand fundamental mechanisms in thiol-based redox signaling, and 2) novel chem- ical matter that can be mined as a source of small-molecule probes and as starting points for drug discovery.