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学科主题物理化学
Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method
Cheng, Kai1,4; Chen, Rui1,2; Seebun, Deeptee2; Ye, Mingliang1; Figeys, Daniel2,3; Zou, Hanfa1; Ye ML(叶明亮); DanielFigeys
关键词N-glycoproteomics Mass Spectrometry Glycan Structure Software Platform
刊名JOURNAL OF PROTEOMICS
2014-10-14
DOI10.1016/j.jprot.2014.08.006
110期:1页:145-154
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Biochemical Research Methods
研究领域[WOS]Biochemistry & Molecular Biology
关键词[WOS]HYDROPHILIC INTERACTION CHROMATOGRAPHY ; ELECTRON-TRANSFER DISSOCIATION ; MASS-SPECTROMETRY DATA ; PROTEIN GLYCOSYLATION ; GLYCAN STRUCTURE ; CELL-ADHESION ; C-TRAP ; IDENTIFICATION ; COMPLEX ; SEARCH
英文摘要The detailed characterization of site-specific glycosylation requires the identification of glycan composition and specific attachment sites on proteins, which need the identification of intact glycopeptides by mass spectrometry. In this study, we present an analytical and computational strategy for the high throughput characterization of intact N-glycopeptides derived from complex proteome samples. N-glycopeptides were identified using the spectra acquired for intact glycopeptides as well as de-glycopeptides. The Y1 ion (peptide + GlcNAc) was accurately determined from the spectra of intact glycopeptides, and the structure of glycan was then identified by searching a constructed glycan database with calculated molecular weight of glycans and their fragment ions. The peptide sequences of intact glycopeptides were identified by matching the molecular weight calculated from Y1 ion to that of deglycosylated peptides from the same HILIC enrichment and identified by a separated LC-MS/MS analysis. The fully automated software platform integrates all of the above processes involved in the identification of the intact N-glycopeptides. This platform was applied to detailed characterization of site-specific glycosylation in HEK 293T cells, which led to the identification of 2249 unique intact N-glycopeptides. These intact glycopeptides revealed 1769 site-specific N-glycans on 453 glycosylation sites which demonstrated the high heterogeneity of glycosylations.
语种英语
WOS记录号WOS:000345183200012
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被引频次:36[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/144529
专题中国科学院大连化学物理研究所
通讯作者Ye ML(叶明亮); DanielFigeys
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Ottawa, Fac Med, Dept Biochem Microbiol & Immunol, Ottawa Inst Syst Biol, Ottawa, ON K1H 8M5, Canada
3.Univ Ottawa, Fac Sci, Dept Chem, Ottawa, ON K1N 6N5, Canada
4.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
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Cheng, Kai,Chen, Rui,Seebun, Deeptee,et al. Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method[J]. JOURNAL OF PROTEOMICS,2014,110(1):145-154.
APA Cheng, Kai.,Chen, Rui.,Seebun, Deeptee.,Ye, Mingliang.,Figeys, Daniel.,...&DanielFigeys.(2014).Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method.JOURNAL OF PROTEOMICS,110(1),145-154.
MLA Cheng, Kai,et al."Large-scale characterization of intact N-glycopeptides using an automated glycoproteomic method".JOURNAL OF PROTEOMICS 110.1(2014):145-154.
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