METHODS FOR AUTOMATED SYNTHESES OF OLIGOSACCHARIDES AND LECTIN TAGS

描述（由申请人提供）：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一直受到明确定义的聚糖结构的阻碍。该项目的总体目标是响应RFA-GM-09-005“扩展碳水化合物的化学空间”，开发几种关键聚糖构建块的快速途径，开发自动化溶液相合成不仅包含常见的而且特别困难的糖苷键的生物学重要结构的方法，并开发一种新工具，使用这些合成的聚糖来识别低聚糖的蛋白质伴侣，例如，致病菌表面。该项目的具体目标是：1)开发自动合成β -葡糖苷和最具挑战性的糖苷键之一- -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糖的自动合成；2)开发自动合成秀丽隐杆线虫细胞表面高支链糖的方法，这些糖是各种细菌病原体感染的关键；3)开发了自动合成含糖醛酸重复寡糖的方法，并将这些方法应用于某些细菌荚膜多糖和糖胺聚糖的合成；4)开发一种新的固相交联试剂，将这些自动合成平台的聚糖产物结合在一起，用质谱法鉴定碳水化合物结合蛋白。

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.