Chemical Probes to Study Methionine Redox Biology

研究蛋氨酸氧化还原生物学的化学探针

• 批准号: 10379768
• 负责人: Christopher J. Chang
• 金额: $ 8.96万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10379768
• 关键词: Antioxidants; Biochemical; Biological Assay; Biology; Cell Proliferation; Cell model; Cells; Chemicals; Chemistry; Disease; Enzymes; Homeostasis; Image; Label; Ligation; Link; Longevity; Methionine; Modeling; Monitor; Nerve Degeneration; Neurodegenerative Disorders; Oxidation-Reduction; Oxidative Stress; Pathologic; Pathology; Physiological; Physiological Processes; Physiology; Proteins; Proteome; Proteomics; Reagent; Regulation; Signal Transduction; Site; Specificity; Thrombosis; Time; Vascular Diseases; adduct; base; cancer cell; chemoproteomics; design; fundamental research; imaging approach; imaging probe; innovation; methionine sulfoxide; methionine sulfoxide reductase; oxaziridine; oxidation; predictive modeling; protein function; public health relevance; research study; tool; tumor progression

Project Summary/Abstract Protein methionine residues are emerging as essential redox sites for physiological processes spanning signal transduction, antioxidant defense, and regulation of protein function. At the same time, aberrant elevations in methionine oxidation induced by oxidative stress and/or inactivation of methionine sulfoxide reductase enzymes can contribute to neurodegenerative and vascular diseases, life span, and cancer progression. To help decipher contributions of methionine redox physiology and pathology, we are developing new chemical reagents to selectively label reactive methionine sites in proteins. The scientific premise is that providing access to a modular chemical toolbox for methionine bioconjugation will help unveil new functional methionine sites in proteins to further understanding of physiological and pathological contributions of methionine oxidation and reduction. Selective methionine bioconjugation using redox-based probes that emulate the innate oxidative chemistry of these residues offers chemical innovation, while demonstrating scientific rigor through the combined use of diverse and complementary synthetic, modeling, biochemical, proteomic, and imaging approaches to provide general tools for probing the reactive methionine landscape. Specifically, we seek to (1) design and synthesize robust and highly specific oxaziridine probes by establishing a predictive model for tuning selectivity and stability of methionine adducts, (2) identify oxidatively-sensitive methionine sites and biochemically characterize specific protein targets involved in reversible redox regulation by chemoproteomic comparison of cell models with or without the methionine sulfoxide reductase eraser protein on a proteome- wide scale, and (3) develop new imaging probes based on proximity ligation assays to monitor methionine- dependent redox status of selected protein targets in cells.

项目摘要/摘要 蛋白蛋氨酸残基正在成为跨越信号的生理过程的必需氧化还原位点 转导，抗氧化剂防御和蛋白质功能的调节。同时，异常高程 蛋氨酸氧化是由氧化应激诱导的和/或抗蛋氨酸硫氧化物还原酶灭活的 酶可以有助于神经退行性和血管疾病，生命跨度和癌症进展。到 帮助破译蛋氨酸氧化还原生理和病理的贡献，我们正在开发新化学 在蛋白质中有选择性标记反应性蛋氨酸位点的试剂。科学前提是提供 访问蛋氨酸生物结合的模块化化学工具箱将有助于揭露新的功能性甲硫酸 蛋白质中的位点，以进一步了解蛋氨酸氧化的生理和病理贡献 和减少。选择性蛋氨酸生物结合使用基于氧化还原的探针，这些探针模仿先天氧化 这些残留物的化学反应提供了化学创新，同时通过 多种和互补合成，建模，生化，蛋白质组学的结合使用 提供通用工具来探测反应性蛋氨酸景观的方法。具体来说，我们寻求（1） 通过建立一个预测模型来设计和合成坚固且高度特异 调整蛋氨酸加合物的选择性和稳定性，（2）识别氧化敏感的蛋氨酸位点和 生化表征了参与化学蛋白质组学可逆氧化还原调节的特定蛋白质靶标 在蛋白质组上的带有或不含甲硫氨酸还原酶橡皮橡胶蛋白的细胞模型的比较 大规模和（3）基于接近连接测定法开发新的成像探针，以监测蛋氨酸 - 细胞中选定蛋白靶标的依赖性氧化还原状态。