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Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples
Wang, Fangjun; Han, Guanghui; Yu, Zhiyuan; Jiang, Xinning; Sun, Shutao; Chen, Rui; Ye, Mingliang; Zou, Hanfa; Zou HF(邹汉法)
关键词Fractionating Human Liver Phosphoproteome Strong Anion-exchange Trap Column
刊名JOURNAL OF SEPARATION SCIENCE
2010-07-01
ISSN1615-9306
DOI10.1002/jssc.200900718
33期:13页:1879-1887
收录类别SCI
文章类型Article
部门归属18
项目归属1809
产权排名1;1
WOS标题词Science & Technology ; Physical Sciences
类目[WOS]Chemistry, Analytical
研究领域[WOS]Chemistry
关键词[WOS]SPECTROMETRY-BASED PROTEOMICS ; ION AFFINITY-CHROMATOGRAPHY ; LC-MS/MS ANALYSIS ; PHOSPHORYLATED PEPTIDES ; MASS-SPECTROMETRY ; POSTTRANSLATIONAL MODIFICATIONS ; MAGNETIC NANOPARTICLES ; LIQUID-CHROMATOGRAPHY ; SHOTGUN PROTEOMICS ; MOUSE-LIVER
英文摘要It is one of the key issues to develop powerful fractionating method to increase the identification of the low-abundance phosphopeptides. In this study, a semi-online 2-D LC separation strategy based on three-step fractionation of the enriched peptides on strong anion-exchange trap column was developed. It was demonstrated that the sensitivity and phosphoproteome coverage obtained by this fractionating method with strong anion-exchange trap column is much higher than those by the conventional methods based on C18 trap column. In addition, when the same amount of sample was loaded, the number of identified phosphopeptides had increased 108%. Combination of this three-step fractionation method with RPLC-MS/MS analysis by 300 min RP-gradient separation was applied to phosphoproteome analysis of human liver proteins, and 853 unique phosphopeptides was positively identified from 500 mu g tryptic digest of human liver proteins. After three cycles' consecutive analyses, 1554 unique phosphopeptides and 1566 phosphorylated sites were totally identified from 735 phosphorylated proteins at a false discovery rate of < 1% in about 54h of analysis time.
语种英语
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WOS记录号WOS:000280163900001
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被引频次:15[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/321008/103147
专题中国科学院大连化学物理研究所
通讯作者Zou HF(邹汉法)
作者单位Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
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Wang, Fangjun,Han, Guanghui,Yu, Zhiyuan,et al. Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples[J]. JOURNAL OF SEPARATION SCIENCE,2010,33(13):1879-1887.
APA Wang, Fangjun.,Han, Guanghui.,Yu, Zhiyuan.,Jiang, Xinning.,Sun, Shutao.,...&邹汉法.(2010).Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples.JOURNAL OF SEPARATION SCIENCE,33(13),1879-1887.
MLA Wang, Fangjun,et al."Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples".JOURNAL OF SEPARATION SCIENCE 33.13(2010):1879-1887.
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