Characterization and Quantification of Highly Sulfated Glycosaminoglycan Isomers by Gated-Trapped Ion Mobility Spectrometry Negative Electron Transfer Dissociation MS/MS.
Characterization and Quantification of Highly Sulfated Glycosaminoglycan Isomers by Gated-Trapped Ion Mobility Spectrometry Negative Electron Transfer Dissociation MS/MS.
复制标题
通过门控离子迁移谱负电子转移解离 MS/MS 对高度硫酸化的糖胺聚糖异构体进行表征和定量。
DOI：
10.1021/acs.analchem.8b05283
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发表时间：
2019
影响因子：
7.4
通讯作者：
Lin,Cheng
中科院分区：
文献类型：
--
作者：
Wei,Juan;Wu,Jiandong;Tang,Yang;Ridgeway,MarkE;Park,MelvinA;Costello,CatherineE;Zaia,Joseph;Lin,Cheng
糖胺聚糖（GAGs）在许多生物过程中起着至关重要的作用，是天然存在的具有巨大结构异质性的复杂多糖混合物，包括许多结构异构体。质谱分析GAG异构体，特别是高硫酸肝素（Hep）和硫酸肝素（HS），具有挑战性，因为它们的结构相似，分析过程中容易失去硫。本研究表明，高硫酸化的Hep/HS异构体可以通过栅极-捕获离子迁移谱法（gate - tims）分辨，而硫损失可以忽略不计。随后对tims分离的Hep/HS异构体进行负电子转移解离（NETD）串联质谱（MS/MS）分析，产生广泛的糖苷和交叉环片段，以确定异构体的区分和结构阐明。门控tims - MS分析提供的高迁移率分辨率和保存不稳定的硫修饰也允许对高硫化肝素异构体进行相对定量。这些结果表明，gate - tims - netd MS/MS方法可用于高硫酸盐Hep/HS化合物的定性和定量分析，这是其他技术无法实现的。
Glycosaminoglycans (GAGs) play vital roles in many biological processes and are naturally present as complex mixtures of polysaccharides with tremendous structural heterogeneity, including many structural isomers. Mass spectrometric analysis of GAG isomers, in particular highly sulfated heparin (Hep) and heparan sulfate (HS), is challenging because of their structural similarity and facile sulfo losses during analysis. Herein, we show that highly sulfated Hep/HS isomers may be resolved by gated-trapped ion mobility spectrometry (gated-TIMS) with negligible sulfo losses. Subsequent negative electron transfer dissociation (NETD) tandem mass spectrometry (MS/MS) analysis of TIMS-separated Hep/HS isomers generated extensive glycosidic and cross-ring fragments for confident isomer differentiation and structure elucidation. The high mobility resolution and preservation of labile sulfo modifications afforded by gated-TIMS MS analysis also allowed relative quantification of highly sulfated heparin isomers. These results show that the gated-TIMS-NETD MS/MS approach is useful for both qualitative and quantitative analysis of highly sulfated Hep/HS compounds in a manner not possible with other techniques.