糖蛋白质结构生物质谱鉴定技术研究

Glycosylation has significant biological function. The structure of glycoprotein plays important role in its functionality. Bio-mass spectrometry with soft ionization is widely used in the structure analysis of glycoproteins. However, due to the highly complexity of glycoprotein, especially the heterogeneity of glycans, the identification of glycoconjugates structure and its function has long suffered from limitation of analytical techniques and thus slowed its development. Developing mass spectrumetry based techniques for glycoprotein analysis as well as realization of intelligent analysis of glycan structure remains one of the most important and difficult tasks.On the basis of previous work,our group will focus on developing Nano-ESI/MALDI based tandem mass spectrometry techniques for glycan branch structrue analysis; we will develop high definition and high sensitivity mass spectrumetry techologies using HCD together with CID and ETD for glycan sequence analysis; We will also generate expert knowledge based sub-databases for glycoprotein searching and feedback control strategy of mass spectrumetry parameters,aiming at solving the problem of semi-automatic deciphering of glycoproteins.

糖基化修饰具有重要的生物学意义，糖蛋白的结构对其功能有着重要的影响。基于软电离技术的生物质大分子谱检测技术在糖基化蛋白质的结构鉴定中有着十分重要的应用。但受制于糖蛋白的高度复杂性特别是糖链结构微观不均一性，糖复合物的结构与功能的研究长期以来受到分析技术的限制而进展缓慢。解决糖蛋白的质谱鉴定特别是N-糖基化修饰谱的结构鉴定和智能解析仍然是目前研究的重点和难点。我们课题组在前期工作的基础上，将重点发展Nano-ESI/MALDI为离子源的多级串级质谱技术，解决糖链分支结构的问题；发展HCD为主结合CID和ETD的高分辨率、高灵敏的质谱技术，解决支链的糖基序列问题；发展基于专家知识的糖蛋白子库产生/搜索和质谱仪器参数的反馈控制，以解决糖蛋白鉴定的结构半自动解析问题。

糖基化修饰具有重要的生物学意义，糖蛋白的结构对其功能有着重要的影响。基于软电离技术的生物质大分子谱检测技术在糖基化蛋白质的结构鉴定中有着十分重要的应用。但受制于糖蛋白的高度复杂性特别是糖链结构微观不均一性，糖复合物的结构与功能的研究长期以来受到分析技术的限制而进展缓慢。解决糖蛋白的质谱鉴定特别是N-糖基化修饰谱的结构鉴定和智能解析仍然是目前研究的重点和难点。本项目重点开发了基于色谱-质谱的高通量完整糖肽、糖链鉴定技术；开发了相应的鉴定流程和搜索软件，包括完整糖肽和糖链鉴定软件，解决了基于质谱糖链、糖肽解析高通量解析问题。