Mass Spectrometry of Metal-Coordinated Oligosaccharides

描述（由申请人提供）：本提案中概述的研究涉及使用傅立叶变换离子回旋共振（FTICR）和离子阱质谱法结合碰撞诱导解离（CID）和红外多光子解离（IRMPD）表征碳水化合物结构。在第一部分中，具体目标包括：官能团变体的位点特异性分析，特别强调a）确定寡糖上的磷酸化位点，B）唾液酸部分的稳定性和位点特异性测定，c）硫酸化的特异性连接位点的测定和GAG硫酸乙酰肝素的序列分析。该提案的第二部分涉及用于鉴定阿拉伯呋喃糖中的结构和连接位置的方法开发以及用于表征从分枝杆菌中提取的分枝菌酰阿拉伯半乳聚糖（mAG）和脂阿拉伯甘露聚糖（LAM）的方法的实施。大多数的一般协议建议纳入使用金属离子或金属配体系统，以实现结构解析的目标。与吉布森、Lowary、考克斯和Sasisekharan博士合作，将方法应用于生物分离和相关碳水化合物，方法开发是双尾的。

DESCRIPTION (provided by applicant): Research outlined in this proposal involves the use of fourier transform ion cyclotron resonance (FTICR) and ion trap mass spectrometry in combination with collision induced dissociation (CID) and infrared multiphoton dissociation (IRMPD) for the characterization of carbohydrate structure. In the first portion specific goals include: Site specific analysis of functional group variants with particular emphasis on a) determining phosphorylation sites on oligosaccharides, b) stabilization of sialic acid moieties and site specific determination, c) determination of specific linkage sites of sulfation and sequence analysis of GAG heparan sulfate. The second part of this proposal addresses both the methods development for identifying structure and linkage position in arabinofuranoses as well as implementation of the method for characterization of mycolyl arobinogalactan (mAG) and lipoarabinomannan (LAM) extracted from mycobacteria. Most of the general protocol proposed incorporates the use of metal ions or metal-ligand systems to achieve the goal of structural elucidation. Methods development is dove-tailed with application of the methods to biologically isolated and relevant carbohydrates as collaborations with Drs. Gibson, Lowary, Cox and Sasisekharan.