METHODS FOR AUTOMATED SYNTHESES OF OLIGOSACCHARIDES AND LECTIN TAGS

描述(申请人提供)：碳水化合物是理解和最终调节宿主-病原体相互作用的关键，但这些相互作用的研究因获得明确定义的多糖结构而受到阻碍。该项目的总体目标是响应RFA-GM-09-005“扩大碳水化合物的化学空间”，开发通往几个关键的糖链的快速路线，开发自动化的溶液相合成生物重要结构的方法，这些结构不仅包含常见的糖苷键，而且还包含特别困难的糖苷键，并开发一种新的工具，使用这些合成的糖链来识别例如病原菌表面的寡糖的蛋白质伙伴。拟议项目的具体目标是：1)开发自动合成β-葡萄糖苷和最具挑战性的糖苷键之一-β-甘露糖苷的方法，并将这些方法应用于真菌相关葡聚糖和甘露聚糖的自动合成；2)开发自动合成线虫线虫细胞表面的高度分枝糖的方法，这些糖是各种细菌病原体感染的关键；3)开发含有糖醛酸的重复低聚糖的自动合成方法，并将这些方法应用于某些细菌被膜多糖和共聚糖的合成；以及4)开发一种新的固相交联剂，该交联剂可以结合自动化合成平台的这些糖链产物，以用于碳水化合物结合蛋白的质谱分析。 公共卫生相关性：碳水化合物是宿主-病原体相互作用的关键，但他们的研究一直受到限制，因为很难获得纯寡糖。该项目的总体目标是响应RFA-GM-09-005“扩大碳水化合物的化学空间”，开发通往几个关键碳水化合物构建块的快速路线，开发自动化的溶液相合成生物重要结构的方法，这些结构不仅包含常见的糖苷键，而且还包含特别困难的糖苷键，并开发一种新的工具，使用这些合成糖聚糖来识别低聚糖的蛋白质伙伴。

DESCRIPTION (provided by applicant): Carbohydrates are key to understanding and ultimately modulating host-pathogen interactions, but the study of these interactions has been hampered by access to well-defined glycan structures. The overall goals of this project, in response to RFA-GM-09-005 "Expanding the Chemical Space of Carbohydrates," are to develop quick routes to several key glycan building blocks, develop methods for the automated solution-phase synthesis of biologically important structures that contain not only common but also particularly difficult glycosidic linkages, and to develop a new tool using these synthetic glycans to identify protein partners of oligosaccharides on, for example, the surface of pathogenic bacteria. The specific aims of the proposed project are to: 1) develop methods for the automated syntheses of beta-glucosides and one of the most challenging glycosidic linkages-beta-mannosides-and apply these methods to the automated syntheses of fungally-associated glucans and mannans; 2) develop methods for the automated synthesis of the highly branched sugars found on the cell surface of the nematode C. elegans that are key to infectivity by various bacterial pathogens; 3) develop methods for the automated synthesis of uronic acid-containing repeating oligosaccharides and apply these methods to the synthesis of certain bacterial capsular polysaccharides and glycosaminoglycans; and 4) develop a new solid-phase-based cross-linking reagent that can incorporate these glycan products of the automated synthesis platform to identify carbohydrate binding proteins by mass spectrometry. PUBLIC HEALTH RELEVANCE: Carbohydrates are key to host-pathogen interactions, but their study has been limited because pure oligosaccharides are hard to obtain. The overall goals of this project, in response to RFA- GM-09-005 "Expanding the Chemical Space of Carbohydrates," are to develop quick routes to several key carbohydrate building blocks, develop methods for the automated solution-phase synthesis of biologically important structures that contain not only common but also particularly difficult glycosidic linkages, and to develop a new tool using these synthetic glycans to identify protein partners of oligosaccharides.