NOVEL APPROACHES TO PREVENTING URINARY TRACT INFECTION IN WOMEN

一种单唾液酸神经节苷脂，被证明能与 从人肾组织中提取和纯化了致尿路病原性大肠杆菌。 神经节苷脂的完整一级结构通过 一维和二维核磁共振氢谱的结合，电喷雾电离 质谱学与电喷雾电离/碰撞 全甲基化完整的诱导解离MS/MS(ESI-MS/CID-MS) 化合物，GC/EI-MS进行甲基化连锁分析。1H-核磁共振 在600 MHz下对800(g，共 鞘糖脂，氢交换并溶解于DMSO-d6/2% D2O。在初步积累了一维1H-核磁共振谱后， 通过以下方法对多糖和神经酰胺的结构特征进行评估 反常质子与其他结构报告基团的比较 与已知的神经鞘糖脂的同源性。标准的2-D系列 然后进行实验(DQF-COSY、TOCSY、NOESY)以指定为 尽可能多的共鸣，以建立身份和一个埃里克形式 所有单糖残基，并建立序列和尽可能多的 通过糖苷间偶极相互作用尽可能地连接位点。 然后用Hakomori方法对等分(100(G))进行全甲基化， 一半进行ESI-MS和-MS/MS分析(SCHEX API-III) 确认了糖链序列和神经酰胺特性，另一半是 解聚和衍生化为部分甲基化的阿迪醇乙酸酯 (PMAA)用于GC-MS分析(惠普5890 GC/5970 MSD，DB-5 列)。通过保留时间和EI对PMAAs进行了鉴定 与已知标准比较的碎片化模式；这些证实 所有己糖和连接形式的同一性和连接性 神经节苷脂中的2-脱氧-2-乙酰氨基己糖残留量。脂肪酸 甲烷裂解产物甲酯的GC-MS分析 神经节苷脂用于确认神经酰胺类型的分布 在这个准备过程中。描述这项工作的手稿已经被 已提交供出版。

A monosialosyl ganglioside shown to bind specifically to uropathogenic E. coli was extracted and purified from human kidney. The complete primary structure of the ganglioside was elucidated by a combination of 1- and 2-D 1H-NMR spectroscopy, electrospray ionization mass spectroscopy (ESI-MS) and electrospray ionization/collision induced dissociation MS/MS (ESI-MS/CID-MS) of the permethylated intact compound, and methylation linkage analysis by GC/EI-MS. 1H-NMR spectroscopy at 600 MHz was carried out on 800 (g of glycosphingolipid, deuterium exchanged and dissolved in DMSO-d6/2% D2O. Following preliminary accumulation of a 1-D 1H-NMR spectrum, an assessment of glycan and ceramide structural features was made via comparison of anomeric proton and other structural reporter groups with those of known glycosphingolipids. A standard series of 2-D experiments (DQF-COSY, TOCSY, NOESY) was then performed to assign as many resonances as possible, to establish identity and anom eric form of all monosaccharide residues, and to establish sequence and as many linkage sites as possible via interglycosidic dipolar interactions. An aliquot (100 (g) was then permethylated by the Hakomori method, one half subjected to ESI-MS and -MS/MS analysis (Sciex API-III) to confirm glycan sequence and ceramide features, and the other half was depolymerized and derivatized to partially methylated alditol acetates (PMAAs) for GC-MS analysis (Hewlett-Packard 5890 GC/5970 MSD, DB-5 column). Identification of PMAAs was made by retention times and EI fragmentation patterns compared with known standards; these confirmed both the identity and linkage form of all hexose and 2-deoxy-2-acetamidohexose residues in the ganglioside. Fatty acid analysis by GC-MS of the methyl esters obtained from methanolysis of the ganglioside was used to confirm the distribution of ceramide types in this preparation. A manuscript describing this work has been submitted for publication.