修饰蛋白质组学高效分离鉴定新技术新方法研究

Post-translational modifications (PTMs) of proteins play very important roles in biological and physiological functions of proteins in living systems. Thus charaterization and identification of PTMs of proteins is one of key issues to understanding the roles of proteins in living systems. Even though the big progress has been achieved on the large-scale (proteome level) charaterization and identification of PTMs of proteins, however it is still at early stage for methods and technology in charaterization and identification of PTMs of proteins now. In this project, the new methods and technology on the proteome analysis of PTMs proteins will be developed with improving the specificity and recovery of enrichment materials, detection sensitivity, identification coverage, and analysis through-put. The main research directions will focus on the enrichment of PTMs peptides and proteins with nano/mesoporous materials, and the integration platform by on-line combination of effective enrichment with adsorbents, immobilized enzyme micro-reactor, solid-phase isotope labeling with nano LC-MS/MS systems. The developed system will applied to highly sensitive proteome analysis of PTMs proteins in rarely biological samples, such as protein complexes and protein extracts from brain tissues.

蛋白质翻译后修饰的种类、数量、结构鉴定对于蛋白质生理功能的诠释有着极其重要的作用。针对目前生物样品修饰蛋白质组学蛋白质组定性、定量分析存在的技术瓶颈，本项目拟开展高选择性、高准确度、高重复性、高灵敏度和高通量的蛋白质翻译后修饰分离鉴定和定量分析新技术新方法的研究。所涉及的主要研究内容包括基于介孔材料的修饰蛋白质/多肽的高选择性富集新方法，微量生物样品的酶解、修饰多肽富集、同位素固相原位标记和质谱连用集成化技术和方法的研究。这些技术和方法的突破有望大大提升修饰蛋白质组学分析的灵敏度、分析通量和定量准确性，实现微/纳升级样品修饰蛋白质组学的规模化定性定量分析，为重大疾病的早期诊断以及药物先导物的高通量筛选等生命科学前沿研究提供新的技术体系和平台。

针对目前生物样品修饰蛋白质组学定性、定量分析存在的技术瓶颈，结合申请者在生物分离分析新技术和新材料科研工作中所形成的基础和积累，开展了蛋白质翻译后修饰分离鉴定和定量分析新技术新方法的创新性研究，有效提高了我国在蛋白质修饰谱方面的研究水平，形成一个具有国际一流水平的蛋白质修饰组新技术新方法的研究团队。相关研究成果发表学术论文86 篇。主要研究成果包括磷酸化蛋白质组的规模化分析平台，糖基化蛋白质组的规模化分析平台，修饰酶与底物相互作用的分析新方法；以及相关新技术在生物医学研究中的应用等。这些技术和方法的突破大大提升了修饰蛋白质组学分析的灵敏度、分析通量和定量准确性，实现了微/纳升级样品修饰蛋白质组学的规模化定性定量分析，为重大疾病的早期诊断以及药物先导物的高通量筛选等生命科学前沿研究提供新的技术体系和平台。