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学科主题: 物理化学
题名: N-Glycosylation Site Analysis of Proteins from Saccharomyces cerevisiae by Using Hydrophilic Interaction Liquid Chromatography-Based Enrichment, Parallel Deglycosylation, and Mass Spectrometry
作者: Cao, Liwei1;  Yu, Long1;  Guo, Zhimou1;  Shen, Aijin1;  Guo, Yunu1;  Liang, Xinmiao1
通讯作者: 于龙 ;  梁鑫淼
关键词: glycoproteomics ;  Saccharomyces cerevisiae ;  hydrophilic interaction liquid chromatography ;  glycopeptide ;  mass spectrometry ;  yeast
刊名: JOURNAL OF PROTEOME RESEARCH
发表日期: 2014-03-01
DOI: 10.1021/pr401049e
卷: 13, 期:3, 页:1485-1493
收录类别: SCI
合作性质: 
文章类型: Article
部门归属: 18
项目归属: 1803
产权排名: 待补充
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods
资助者: 1,1
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: N-Glycosylation site analysis of baker's yeast Saccharomyces cerevisiae is of fundamental significance to elucidate the molecular mechanism of human congenital disorders of glycosylation (CDG). Here we present a mass spectrometry (MS)-based workflow for the profiling of N-glycosylated sites in S. cerevisiae proteins. In this workflow, proteolytic glycopeptides were enriched by using a hydrophilic material named Click TE-Cys to improve the glycopeptide selectivity and coverage. To enhance the reliability of the identified results, the enriched glycopeptides were subjected to parallel deglycosylation by using two endoglycosidases (i.e., PNGase F and Endo H-f), respectively, prior to LC-MS/MS analysis. On the basis of the workflow, a total of 135 N-glycosylated sites including 6 known, 93 potential, and 36 novel sites were identified and mapped to 79 proteins. Among the novel-type sites, nine sites from eight proteins, which were simultaneously identified via PNGase F and Endo H-f deglycosylation, are believed to possess high confidence. The established workflow, together with the profile of N-glycosylated sites, will contribute to the improvement of S. cerevisiae model for revealing the pathogenesis of CDG.
关键词[WOS]: CONGENITAL DISORDERS ;  LINKED GLYCOPROTEINS ;  GLYCOPEPTIDE-ENRICHMENT ;  SELECTIVE ENRICHMENT ;  SERUM GLYCOPROTEINS ;  IDENTIFICATION ;  YEAST ;  MATRIX ;  LECTIN ;  QUANTIFICATION
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000332756300028
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/119718
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China

Recommended Citation:
Cao, Liwei,Yu, Long,Guo, Zhimou,et al. N-Glycosylation Site Analysis of Proteins from Saccharomyces cerevisiae by Using Hydrophilic Interaction Liquid Chromatography-Based Enrichment, Parallel Deglycosylation, and Mass Spectrometry[J]. JOURNAL OF PROTEOME RESEARCH,2014,13(3):1485-1493.
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