Assignment of Saccharide Identities through Analysis of Oxonium Ion Fragmentation Profiles in LC MS/MS of Glycopeptides
Assignment of Saccharide Identities through Analysis of Oxonium Ion Fragmentation Profiles in LC MS/MS of Glycopeptides
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DOI：
10.1021/pr500898r
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发表时间：
2014-12-01
影响因子：
4.4
通讯作者：
Nilsson, Jonas
中科院分区：
文献类型：
--
作者：
Halim, Adnan;Westerlind, Ulrika;Nilsson, Jonas
蛋白质糖基化在多种生物过程的调节中发挥着关键作用，确定聚糖结构-功能关系对于更好地理解这些事件非常重要。然而，聚糖和糖肽结构异构体的表征仍然具有挑战性，并且通常依赖于保守的生物合成途径。在使用碰撞诱导解离 (CID) 的液相色谱-串联质谱 (LC-MS/MS) 糖蛋白组学分析中，含有 N-乙酰己糖胺 (HexNAc) 残基的糖氧鎓离子很突出。通过对合成和天然衍生的 N- 和 O- 糖肽的束型 CID 光谱和离子阱 CID 光谱分析，我们发现末端被 GalNAca1-O-、GlcNAc beta 1-O-、Gal beta 3GalNAca1-O-、Gal beta 4GlcNAc beta-O- 和 Gal beta 3GlcNAc beta-O- 结构取代的糖肽之间的氧鎓离子断裂模式具有特征性差异。因此，此类糖肽的氧鎓离子断裂谱的差异可用于区分这些聚糖结构，并且在基于 LC-MS/MS 的糖蛋白质组研究中可能很重要。
Protein glycosylation plays critical roles in the regulation of diverse biological processes, and determination of glycan structure-function relationships is important to better understand these events. However, characterization of glycan and glycopeptide structural isomers remains challenging and often relies on biosynthetic pathways being conserved. In glycoproteomic analysis with liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID), saccharide oxonium ions containing N-acetylhexosamine (HexNAc) residues are prominent. Through analysis of beam-type CID spectra and ion trap CID spectra of synthetic and natively derived N- and O-glycopeptides, we found that the fragmentation patterns of oxonium ions characteristically differ between glycopeptides terminally substituted with GalNAca1-O-, GlcNAc beta 1-O-, Gal beta 3GalNAca1-O-, Gal beta 4GlcNAc beta-O-, and Gal beta 3GlcNAc beta-O- structures. The difference in the oxonium ion fragmentation profiles of such glycopeptides may thus be used to distinguish among these glycan structures and could be of importance in LC-MS/MS-based glycoproteomic studies.