De Novo Glycan Sequencing by Electronic Excitation Dissociation and Fixed-Charge Derivatization,Analytical Chemistry

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De Novo Glycan Sequencing by Electronic Excitation Dissociation and Fixed-Charge Derivatization
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-02-14 00:00:00 , DOI: 10.1021/acs.analchem.7b04077
Yang Tang _{1,

2} , Yi Pu _{1,

2} , Jinshan Gao ₃ , Pengyu Hong ₄ , Catherine E. Costello _{1,

2} , Cheng Lin ₁

Affiliation

Detailed glycan structural characterization is frequently achieved by collisionally activated dissociation (CAD) based sequential tandem mass spectrometry (MSⁿ) analysis of permethylated glycans. However, it is challenging to implement MSⁿ (n > 2) during online glycan separation, and this has limited its application to analysis of complex glycan mixtures from biological samples. Further, permethylation can reduce liquid chromatographic (LC) resolution of isomeric glycans. Here, we studied the electronic excitation dissociation (EED) fragmentation behavior of native glycans with a reducing-end fixed charge tag and identified key spectral features that are useful for topology and linkage determination. We also developed a de novo glycan sequencing software that showed remarkable accuracy in glycan topology elucidation based on the EED spectra of fixed charge-derivatized glycans. The ability to obtain glycan structural details at the MS² level, without permethylation, via a combination of fixed charge derivatization, EED, and de novo spectral interpretation, makes the present approach a promising tool for comprehensive and rapid characterization of glycan mixtures.

中文翻译：

通过电子离解和固定电荷衍生化从头进行聚糖测序

通常通过基于碰撞活化解离（CAD）的全甲基化聚糖的顺序串联质谱分析（MS ⁿ）分析来实现详细的聚糖结构表征。但是，实施MS ⁿ（n> 2）在在线聚糖分离过程中，这限制了它在分析生物样品中复杂的聚糖混合物中的应用。此外，全甲基化可降低异构聚糖的液相色谱（LC）分离度。在这里，我们研究了具有还原端固定电荷标签的天然聚糖的电子激发解离（EED）片段化行为，并确定了可用于拓扑和连锁确定的关键光谱特征。我们还开发了从头开始的聚糖测序软件，该软件基于固定电荷衍生化聚糖的EED光谱，在糖类拓扑结构阐明中显示出显着的准确性。在MS ²上获得聚糖结构细节的能力通过固定电荷衍生化，EED和从头光谱解释的结合，在没有过甲基化的情况下获得高水平的甲基化，使得本发明的方法成为用于聚糖混合物的全面和快速表征的有前途的工具。

更新日期：2018-02-14

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