Chemical Probes to Study Methionine Redox Biology

项目总结/摘要 蛋白质甲硫氨酸残基是跨信号转导生理过程的重要氧化还原位点 转导、抗氧化防御和调节蛋白质功能。与此同时， 氧化应激和/或蛋氨酸亚砜还原酶失活诱导的蛋氨酸氧化 酶可导致神经退行性疾病和血管疾病、寿命和癌症进展。到 为了帮助破译蛋氨酸氧化还原生理学和病理学贡献，我们正在开发新的化学物质， 选择性标记蛋白质中的反应性甲硫氨酸位点的试剂。科学的前提是， 获得蛋氨酸生物结合的模块化化学工具箱将有助于揭示新的功能性蛋氨酸 进一步了解蛋氨酸氧化的生理和病理作用 和减少。使用基于氧化还原的探针的选择性甲硫氨酸生物缀合 这些残留物的化学性质提供了化学创新，同时通过 多种互补的合成、建模、生物化学、蛋白质组学和成像的组合使用 提供用于探测反应性甲硫氨酸景观的通用工具的方法。具体而言，我们寻求（1） 通过建立预测模型，设计和合成稳健的和高度特异性的氧氮丙啶探针， 调节甲硫氨酸加合物的选择性和稳定性，（2）鉴定氧化敏感的甲硫氨酸位点， 通过化学蛋白质组学对参与可逆氧化还原调节的特定蛋白质靶标进行生物化学表征 蛋白质组上有或没有甲硫氨酸亚砜还原酶擦除蛋白的细胞模型的比较- 大规模，和（3）开发新的成像探针的基础上邻位连接测定监测蛋氨酸- 细胞中所选蛋白质靶标的依赖性氧化还原状态。

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.