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题名: Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples
作者: Wang, Fangjun1;  Han, Guanghui1;  Yu, Zhiyuan1;  Jiang, Xinning1;  Sun, Shutao1;  Chen, Rui1;  Ye, Mingliang1;  Zou, Hanfa1
通讯作者: 邹汉法
关键词: Fractionating ;  Human liver ;  Phosphoproteome ;  Strong anion-exchange ;  Trap column
刊名: JOURNAL OF SEPARATION SCIENCE
发表日期: 2010-07-01
DOI: 10.1002/jssc.200900718
卷: 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
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.
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
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000280163900001
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/103147
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, Natl Chromatog R&A Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China

Recommended Citation:
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.
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