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学科主题: 物理化学
题名: Identification of phosphopeptides with unknown cleavage specificity by a de novo sequencing assisted database search strategy
作者: Dong, Mingming1, 2;  Ye, Mingliang1;  Cheng, Kai1, 2;  Dong, Jing1;  Zhu, Jun1, 2;  Qin, Hongqiang1, 2;  Bian, Yangyang1, 2;  Zou, Hanfa1
通讯作者: 叶明亮 ;  邹汉法
关键词: De novo sequencing ;  Endogenous phosphopeptide ;  Phosphoproteomics ;  Phosphopeptide identification ;  Technology
刊名: PROTEOMICS
发表日期: 2014-11-01
DOI: 10.1002/pmic.201400268
卷: 14, 期:21-22, 页:2410-2416
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 ;  Biochemistry & Molecular Biology
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: In theory, proteases with broad cleavage specificity could be applied to digest protein samples to improve the phosphoproteomic analysis coverage. However, in practice this approach is seldom employed. This is because the identification of phosphopeptides without enzyme specificity by conventional database search strategy is extremely difficult due to the huge search space. In this study, we investigated the performance of a de novo sequencing assisted database search strategy for the identification of such phosphopeptides. Firstly, we compared the performance of conventional database search strategy and the de novo sequencing assisted database search strategy for the identification of peptides and phosphopeptides without stetting enzyme specificity. It was found that the identification sensitivity dropped significantly for the conventional one while it was only slightly decreased for the new approach. Then, this new search strategy was applied to identify phosphopeptides generated by Proteinase K digestion, which resulted in the identification of 717 phosphopeptides. Finally, this strategy was utilized for the identification of serum endogenous phosphopeptides, which were generated in vivo by different kinds of proteases and kinases, and the identification of 68 unique serum endogenous phosphopepitdes was successfully achieved.
关键词[WOS]: SCALE PHOSPHOPROTEOME ANALYSIS ;  CHROMATOGRAPHY ;  ENRICHMENT ;  PEPTIDOME
语种: 英语
WOS记录号: WOS:000344807600004
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/144419
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, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China

Recommended Citation:
Dong, Mingming,Ye, Mingliang,Cheng, Kai,et al. Identification of phosphopeptides with unknown cleavage specificity by a de novo sequencing assisted database search strategy[J]. PROTEOMICS,2014,14(21-22):2410-2416.
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