METHODS FOR AUTOMATED SYNTHESES OF OLIGOSACCHARIDES AND LECTIN TAGS

描述（由申请方提供）：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受到了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“扩大碳水化合物的化学空间”，开发几种关键聚糖结构单元的快速路线，开发自动化液相合成生物学重要结构的方法，这些结构不仅含有常见的糖苷键，而且含有特别困难的糖苷键，并开发一种新的工具，使用这些合成聚糖来鉴定例如致病细菌表面上寡糖的蛋白质伴侣。该项目的具体目标是：1）开发自动化合成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化合成; 2）开发自动化合成线虫C. 3）开发自动合成含糖醛酸的重复寡糖的方法，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）开发一种新型的固相基交叉-连接试剂，其可以掺入自动化合成平台的这些聚糖产物以通过质谱法鉴定碳水化合物结合蛋白。 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，但由于很难获得纯寡糖，因此对其研究受到限制。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”，开发几种关键碳水化合物结构单元的快速路线，开发用于生物学重要结构的自动化液相合成的方法，这些结构不仅包含常见的，而且还包含特别困难的糖苷键，并开发一种新的工具，使用这些合成聚糖来鉴定寡糖的蛋白质伴侣。

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.