选择性分离和富集代谢标记的糖蛋白

项目名称： 选择性分离和富集代谢标记的糖蛋白

项目编号： No.21205038

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 分析化学

项目作者： 曾盈

作者单位： 湖南师范大学

项目金额： 25万元

中文摘要： 糖蛋白上糖链的分支结构以及表达的密度对细胞-细胞间的信号传递，细胞的复制等细胞功能有着举足轻重的作用。糖蛋白微观结构的改变与多种疾病和癌症的发展和转移有着明显的关系，对岩藻糖基化蛋白和唾液酸化糖蛋白的研究对癌症的早期检测和诊断，癌症的介入治疗都有明显的参考意义。在不对细胞的遗传物质进行操作的情况下，利用表达在细胞表面有化学报告基团的非天然糖，通过特殊的化学反应对其进行修饰。本项目将在前期研究的基础上，以岩藻糖和唾液酸为结构基础，以代谢寡糖工程技术为手段，对活体细胞表面的岩藻糖基化蛋白和唾液酸化糖蛋白进行研究。综合运用磁性纳米粒子，对其进行分离和富集，再使用LC-MS，MS， NMR等技术对其进行分析，分析活体细胞该糖蛋白的微观结构及其变化。本项目将为岩藻糖蛋白的生理作用和临床应用提供科学依据。

中文关键词： Fe3O4 磁性纳米粒子；表面修饰；可裂解型功能基团；生物正交化学；糖蛋白组学

英文摘要： Glycosylation is one of the most common and important post-translational modification of the protein, and plays crucial role in many biological events, involved in cell- cell adhesion, recognition and signaling. The diversity and complexity of the micro-structures are related to many diseases and tumor metastasis. Heterogeneity hampered the progress toward understanding the functions of the oligosacccharidses. It is a big chanllenge to enrich and separate the glycans and glycoproteins. In this proposal, we develop an new strategy for enrichment and separation of the fucosylted and sialylated glycoprotein.Understanding the microstructure of sialic acid and fucose has enormous potential for understanding the biological functions and treating diseases. This strategy is based on the combination of metabolic oligosaccharides engineering and nano-technology. The aims are 1)designing, synthesizing new unnatural sialic acid/fucose with chemical reportor-aldehyde, 2)incorporating the unnatural sugar into the cell surface by metabolic engineering, 3) synthesizing and characterizing new functionalized magnetic nano-particle, 4)taging the magnetic nanoparticel onto the cell surface by oxime ligation, 5)enriching and separating the taggd glycopretein, releasing and analyzing the glycoproteins. In this application, we propo

英文关键词： Fe3O4 Magnetic nanoparticle；surface modification；cleavable functional group；bio-orthogonal chemistry；glcoproteomics