糖胺聚糖的序列及其与肿瘤蛋白的相互作用位点研究

项目名称： 糖胺聚糖的序列及其与肿瘤蛋白的相互作用位点研究

项目编号： No.21472115

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 环境科学、安全科学

项目作者： 迟连利

作者单位： 山东大学

项目金额： 80万元

中文摘要： 糖胺聚糖是一类结构复杂的酸性直链多糖大分子。生物学研究表明细胞表面和细胞间基质内的硫酸乙酰肝素类糖胺聚糖通过自身结构的变化调控肿瘤细胞的生长、侵袭和转移。然而研究这些糖链的细微结构如何变化以及怎样影响与肿瘤蛋白的相互作用尚缺乏有力的糖分析化学工具。本项目以现代质谱技术为主要手段，建立和创新硫酸乙酰肝素分析的系统方法，包括指纹图谱方法分析组织糖链及其基本组成单元；正、负离子模式下的多级质谱解析关键寡糖的序列；以及共价交联质谱法和氢氘置换质谱法研究硫酸乙酰肝素与靶蛋白的空间相互作用位点。通过对肺癌小鼠模型组织硫酸乙酰肝素糖链变化的研究验证方法体系的有效性并加以完善，发现肿瘤特征糖链并测定其功能域的序列，并研究其如何影响与肿瘤相关靶蛋白的相互作用。本项目的研究结果将为深入理解糖胺聚糖在肿瘤中的作用机制提供方法和手段，为发现新的肿瘤生物标志物和新药靶标提供结构与功能研究基础。

中文关键词： 糖胺聚糖；硫酸乙酰肝素；质谱；结构与功能；肿瘤

英文摘要： Glycosaminoglycans are complicated and highly negatively charged linear polysaccharides. Biological studies showed that the cell-surface and extracellular-matrix heparan sulfate glycosaminoglycans regulated tumor genesis, progression and metastasis by modifying their structures. However, the lack of powerful carbohydrate analytical chemistry tools limited the understanding of the relationship between fine structure changes of heparan sulfate and the effects on their interaction with tumor factors. In this study, we will establish novel and systematic analytical approaches for heparan sulfate using modern mass spectrometry technologies, including fingerprinting methods for intact heparan sulfate chains as well as their building blocks; sequencing methods for critical heparan sulfate oligosaccharides using tandem mass spectrometry in both the positive and the negative modes; and heparan sulfate-protein interaction motifs elucidating methods using covalently cross-linking and hydrogen/deuterium exchanging followed by mass spectrometry analysis. We will then investigate the heparan sulfate in the mouse lung cancer models in order to examine and improve our approaches. Our aim is to find the significant heparan sulfate modifications and reveal their effects on interaction with tumor factors. This project will provide not only novel approaches for further understanding the role of glycosaminoglycans in tumor, but also structural-function evidences for discovering new tumor biomarkers and drug targets.

英文关键词： glycosaminoglycan;heparan sulfate;mass spectrometry;structure function relationship;tumor