Combination of β-elimination and liquid chromatography/quadrupole time-of-flight mass spectrometry for the determination of O-glycosylation sites

被引：28

作者：

Zheng, Yufang ^{[1
]}

Guo, Zhihong ^{[2
]}

Cai, Zongwei ^{[1
]}

机构：

[1] Hong Kong Baptist Univ, Dept Chem, Kowloon, Hong Kong, Peoples R China

[2] Hong Kong Univ Sci & Technol, Dept Chem, Kowloon, Hong Kong, Peoples R China

来源：

TALANTA | 2009年 / 78卷 / 02期

关键词：

O-glycosylation site; Glycopeptide; beta-Elimination/addition; LC/Q-TOF MS; ELECTRON-CAPTURE DISSOCIATION; FIXED-CHARGE DERIVATIZATION; POST-SOURCE DECAY; GLYCOPEPTIDES; LOCALIZATION; PEPTIDES; IDENTIFICATION; DEGLYCOSYLATION; HYDROLYSIS; THREONINE;

D O I：

10.1016/j.talanta.2008.11.026

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

Determination of O-glycosylation sites in glycopeptides was developed by using two model compounds designed from mucin2 tandem repeat motif and erythropoietin. beta-Elimination/addition reaction using dimethylamine on glycosylated site through a Michael-type condensation produced efficient deglycosylation with appropriate chemical modification. The use of dimethylamine was efficient to release the O-linked glycan in a reaction time period of 2-6 h at 55 degrees C. Peptide sequencing was then performed using the liquid chromatography/quadrupole time-of-flight mass spectrometry and MS-MS experiments. Interpretation of fragmentation pathways of the beta-elimination/addition products enabled straightforward recognition of glycosylation site. Compared to the fragmentation of corresponding native peptides, mass shift of-18 Da or +27 Da was clearly observed for the two kinds of beta-elimination/addition products of the glycosylated threonine. Dimethylamine was found to provide higher efficiency of beta-elimination/addition than methylamine and ammonia. (c) 2008 Elsevier B.V. All rights reserved.

引用

页码：358 / 363

页数：6

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