肝脏蛋白可逆S-脂修饰及农药暴露干扰的蛋白质组学研究

Liver not only plays a major role in the metabolism of exogenous chemicals but also is a key organ for protein synthesis, bile production and immune defense. This work is focusing on the newly revealed reversible S-fatty acylation of hepatic proteins. It is aimed to map fatty acylated proteome of zebrafish liver and identify other related protein modifications (such as phosphorylation) as well as activated/deactivated key protein clusters based on Selective Chemical Hydrolysis-Directed Amino Acid Affinity Localization (SCHAAL) in combination with chromatography-mass spectrometry. By using bioactive pesticides as chemical probes, changes in the levels of protein expression and modification under different levels of exposure can be quantified. The final goal of this work is to reveal the functional protein networks of S-fatty acylation and lay foundations for future studies on molecular enzymatic mechanisms. It is expected to provide model biological data for studies of human diseases of public health significance, and discover new sets of molecular biomarkers that can assess pesticide safety.

肝脏既是外源性化学物的主要代谢器官，也是合成蛋白质、分泌胆汁、免疫防御等的关键场所。本项目旨在围绕蛋白质可逆S-脂修饰，建立选择性化学水解导向的氨基酸位点亲和定位法，简称为SCHAAL法（Selective Chemical Hydrolysis-Directed Amino Acid Affinity Localization）,利用色谱-质谱联用分析体系，先导性构建斑马鱼肝脏蛋白质S-脂修饰谱，鉴定与可逆S-脂修饰相关的其它蛋白修饰（如磷酸化修饰），以及S-脂修饰激活/去激活的关键蛋白质群。并以农药活性小分子为探针，研究不同剂量暴露干扰下修饰谱和蛋白质表达谱的动态变化，揭示S-脂修饰相关的功能蛋白质群，为进一步开展分子酶学机制研究奠定基础。本项目的实施预期可为认识人类重大疾病的发生发展提供新型模式生物数据，发现一批具有农药安全预警能力的分子标志物，为农药安全性评价提供新的实验依据。

本项目围绕蛋白质可逆S-脂修饰，建立了选择性化学水解导向的氨基酸位点亲合定位法，简称为SCHAAL法（Selective Chemical Hydrolysis-Directed Amino Acid Affinity Localization）,利用色谱-质谱联用分析体系，先导性分析了模式生物蛋白质S-脂修饰谱，建立了与脂修饰相关的磷酸化蛋白修饰分析方法，为进一步开展分子酶学机制研究奠定了基础。