Sequencing of sulfated oligosaccharides from mucins by liquid chromatography and electrospray ionization tandem mass spectrometry
Sequencing of sulfated oligosaccharides from mucins by liquid chromatography and electrospray ionization tandem mass spectrometry
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DOI：
10.1021/ac000631b
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发表时间：
2000-10-01
影响因子：
7.4
通讯作者：
Hansson, GC
中科院分区：
文献类型：
--
作者：
Thomsson, KA;Karlsson, H;Hansson, GC
作为分析硫酸化粘蛋白衍生寡糖的不同混合物的策略的一部分，已经探索了负离子模式下的液相色谱-电喷雾串联质谱（ESI-MS/MS）。采用氨基键合色谱柱与Q-TOF仪器连接，对猪胃和大肠中的两种硫酸寡糖糖醇混合物进行了分析。九个硫酸化粘蛋白衍生的寡糖糖醇从猪胃进行了广泛的片段化，允许其序列的测定。碎片化产生的初级、二级和三级碎片离子为阐明糖序列和硫酸根基团的定位提供了信息。从一个单一的色谱分析，从猪大肠纯化的28个不同的硫酸化粘蛋白寡糖糖醇的序列进行了解析，揭示了有关突出的核心序列和末端血型型表位的信息。这两种硫酸化低聚糖混合物的分析证明了HPLC-ESI-MS/MS的有用性：生物样品中存在的多种异构硫酸化低聚糖的在线分离，允许识别非衍生化低聚糖序列的信息片段化，以及足以分析从天然来源获得的量的灵敏度。
As part of a strategy for profiling diverse mixtures of sulfated mucin-derived oligosaccharides, liquid chromatography coupled to electrospray ionization tandem mass spectrometry (ESI-MS/MS) in the negative ion mode has been explored. Two mixtures of sulfated oligosaccharide alditols from porcine stomach and large intestine were analyzed by straight phase chromatography using an amino-bonded column connected to a Q-TOF instrument. Nine sulfated mucin-derived oligosaccharide alditols from porcine stomach underwent extensive fragmentation allowing determination of their sequence. The fragmentation generated primary, secondary, and tertiary fragment ions informative for the elucidation of the saccharide sequence and localization of the sulfate group. From a single chromatographic analysis, the sequences of 28 different sulfated mucin oligosaccharide alditols purified from porcine large intestine were elucidated, revealing information concerning prominent core sequences and terminal blood group-type epitopes. Analysis of these two sulfated oligosaccharide mi;mixtures demonstrated the usefialness of HPLC-ESI-MS/MS: the on-line separation of multiple isomeric sulfated oligosaccharides as present in biological samples, informative fragmentation allowing the identification of the sequence of nonderivatized oligosaccharides, and a sensitivity sufficient for the analysis of quantities as obtained from natural sources.