氧化应激下细胞巯基蛋白翻译后修饰的荧光/拉曼/质谱多模态标度

The regulation of protein function and the related signal transduction is achieved by posttranslational modifications (PTMs) of sulfhydryl protein under oxidative stress. Therefore, detecting biomarkers of sulfhydryl proteins’ PTMs efficiently, specifically and dynamically under oxidative stress and exploring the redox signal transduction, protein function and activity regulation, is one of the important research content to solve the key scientific problems of chemical modification and dynamic process of biomolecules. Due to the lack of methods for directly chemical labeling biomarkers of proteins’ PTMs in living cells avoiding the "fake site" interference, we plan to design and screen chemical labeling reactions with high efficiency and high selectivity. Taking advantages of fluorescence/raman/mass spectrometry, we want to develop a multimodal method to label and quantify biomarkers of sulfhydryl proteins’ PTMs induced by oxidative stress. This multimodal strategy will make up the limitations of unitary detecting methods in real-time labeling and tracking of sulfhydryl protein PTMs biomarkers, and realize accurate and highly spatiotemporal detection of PTMs induced by oxidative stress in the living cells. With the aid of this highly sensitive multimodal detection technique, new types of sulfhydryl protein PTMs in the cells will be studied. These studies will provides important research basis for further understanding the regulations of protein function and signal transduction.

巯基蛋白分子翻译后修饰（PTMs）是氧化应激调控细胞信号转导、蛋白功能及活性的重要途径，因此高效、特异和时空动态检测氧化应激下巯基蛋白PTMs标志物，继而探索氧化还原信号转导、蛋白功能及活性调控机制，是生物大分子化学修饰的化学特征与动态过程这一关键科学问题的重要研究内容之一。针对目前细胞内直接法存在标记方法和检测手段过于单一，难免会有“假位点”干扰的缺陷，本项目通过设计和筛选高效率、高选择性的化学标记反应，融合荧光/拉曼/质谱等多种检测技术，多模态标度氧化应激下巯基蛋白PTMs标志物，实现氧化应激下巯基蛋白PTMs标志物准确定量及时空分布的高效检测；并利用高灵敏多模态检测技术探索氧化应激下未知的巯基蛋白PTMs种类，为深入理解细胞中巯基蛋白PTMs及其在信号转导、蛋白功能调控研究提供重要的基础。

氧化应激下，活性分子通过调控蛋白翻译后修饰，影响生理和病理过程。但是由于缺乏原位、准确检测蛋白修饰的研究方法，阻碍了人们进一步理解蛋白功能发挥和对疾病的影响机制。针对这一关键科学问题，本项目聚焦肝损伤和抑郁症两种重大疾病，发展荧光和质谱多模态分析方法，深入研究了氧化应激下活性分子对相关蛋白的翻译后修饰的调控，及其对肝损伤和抑郁症疾病发生发展的影响机制。本项目资助工作在国际学术期刊发表45篇论文（其中1篇为Nat. Commun., 2篇为J. Am. Chem. Soc., 6篇为Angew. Chem. Int. Ed），并基于氧化应激相关工作积累，应邀为Angew. Chem. Int. Ed撰写综述论文1篇，申请国家发明专利15项。项目执行期间，培养博士生6人，硕士生12人。