中国科学院机构知识库网格
Chinese Academy of Sciences Institutional Repositories Grid
Improve the Coverage for the Analysis of Phosphoproteome of HeLa Cells by a Tandem Digestion Approach

文献类型:期刊论文

作者Bian, Yangyang1,2; Ye, Mingliang1; Song, Chunxia1,2; Cheng, Kai1,2; Wang, Chunli1,2; Wei, Xiaoluan1; Zhu, Jun1,2; Chen, Rui1,2; Wang, Fangjun1; Zou, Hanfa1
刊名journal of proteome research
出版日期2012-05-01
卷号11期号:5页码:2828-2837
关键词tandem protease digestion Glu-C phosphorylation sites phosphoproteomics LC-MS/MS kinase motif
ISSN号1535-3893
产权排序1,1
通讯作者叶明亮 ; 邹汉法
英文摘要complete coverage of all phosphorylation sites in a proteome is the ultimate goal for large-scale phosphoproteome analysis. however, only making use of one protease trypsin for protein digestion cannot cover all phosphorylation sites, because not all tryptic phosphopeptides are detectable in ms. to further increase the phosphoproteomics coverage of hela cells, we proposed a tandem digestion approach by using two different proteases. by combining the data set of the first glu-c digestion and the second trypsin digestion, the tandem digestion approach resulted in the identification of 8062 unique phosphopeptides and 8507 phosphorylation sites in hela cells. the conventional trypsin digestion approach resulted in the identification of 3891 unique phosphopeptides and 4647 phosphorylation sites. it was found that the phosphorylation sites identified from the above two approaches were highly complementary. by combining above two data sets, in total we identified 10899 unique phosphopeptides and 11262 phosphorylation sites, corresponding to 3437 unique phosphoproteins with fdr < 1% at peptide level. we also compared the kinase motifs extracted from trypsin, glu-c, or a second trypsin digestion data sets. it was observed that basophilic motifs were more frequently found in the trypsin and the second trypsin digestion data sets, and the acidic motifs were more frequently found in the glu-c digestion data set. these results demonstrated that our tandem digestion approach is a good complement to the conventional trypsin digestion approach for improving the phosphoproteomics analysis coverage of hela cells.
WOS标题词science & technology ; life sciences & biomedicine
学科主题物理化学
类目[WOS]biochemical research methods
研究领域[WOS]biochemistry & molecular biology
关键词[WOS]scale phosphorylation analysis ; mass-spectrometry ; protein-phosphorylation ; phosphopeptide enrichment ; saccharomyces-cerevisiae ; chromatography ; separation ; elastase ; strategy ; sites
收录类别SCI
语种英语
WOS记录号WOS:000303492100017
公开日期2013-10-11
源URL[http://159.226.238.44/handle/321008/118236]  
专题大连化学物理研究所_中国科学院大连化学物理研究所
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.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
推荐引用方式
GB/T 7714
Bian, Yangyang,Ye, Mingliang,Song, Chunxia,et al. Improve the Coverage for the Analysis of Phosphoproteome of HeLa Cells by a Tandem Digestion Approach[J]. journal of proteome research,2012,11(5):2828-2837.
APA Bian, Yangyang.,Ye, Mingliang.,Song, Chunxia.,Cheng, Kai.,Wang, Chunli.,...&Zou, Hanfa.(2012).Improve the Coverage for the Analysis of Phosphoproteome of HeLa Cells by a Tandem Digestion Approach.journal of proteome research,11(5),2828-2837.
MLA Bian, Yangyang,et al."Improve the Coverage for the Analysis of Phosphoproteome of HeLa Cells by a Tandem Digestion Approach".journal of proteome research 11.5(2012):2828-2837.

入库方式: OAI收割

来源:大连化学物理研究所

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