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题名: Large-scale phosphoproteome analysis of human liver tissue by enrichment and fractionation of phosphopeptides with strong anion exchange chromatography
作者: Han, Guanghui1;  Ye, Mingliang1;  Zhou, Houjiang1;  Jiang, Xinning1;  Feng, Shun1;  Jiang, Xiaogang1;  Tian, Ruijun1;  Wan, Dafang2;  Zou, Hanfa1;  Gu, Jianren2
通讯作者: 邹汉法
关键词: human liver ;  phosphopeptide validation ;  phosphoproteome analysis ;  SAX
刊名: PROTEOMICS
发表日期: 2008-04-01
DOI: 10.1002/pmic.200700884
卷: 8, 期:7, 页:1346-1361
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 ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods ;  Biochemistry & Molecular Biology
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: The mixture of phosphopeptides enriched from proteome samples are very complex. To reduce the complexity it is necessary to fractionate the phosphopeptides. However, conventional enrichment methods typically only enrich phosphopeptides but not fractionate phosphopeptides. In this study, the application of strong anion exchange (SAX) chromatography for enrichment and fractionation of phosphopeptides was presented. it was found that phosphopeptides were highly enriched by SAX and majority of unmodified peptides did not bind onto SAX. Compared with Fe3+ immobilized metal affinity chromatography (Fe3+-IMAC), almost double phosphopeptides were identified from the same sample when only one fraction was generated by SAX. SAX and Fe3+-IMAC showed the complementarity in enrichment and identification of phosphopeptides. It was also demonstrated that SAX have the ability to fractionate phosphopeptides under gradient elution based on their different interaction with SAX adsorbent. SAX was further applied to enrich and fractionate phosphopeptides in tryptic digest of proteins extracted from human liver tissue adjacent to tumorous region for phosphoproteome profiling. This resulted in the highly confident identification of 274 phosphorylation sites from 305 unique phosphopeptides corresponding to 168 proteins at false discovery rate (FDR) of 0.96%.
关键词[WOS]: ION AFFINITY-CHROMATOGRAPHY ;  TANDEM MASS-SPECTROMETRY ;  PROTEIN IDENTIFICATION TECHNOLOGY ;  TITANIUM-DIOXIDE MICROCOLUMNS ;  GROWTH-FACTOR RECEPTOR ;  IN-VIVO ;  TYROSINE PHOSPHORYLATION ;  SACCHAROMYCES-CEREVISIAE ;  SIGNALING PATHWAYS ;  MEMBRANE-PROTEINS
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000254986200002
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/99953
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, Dalian 116023, Peoples R China
2.Shanghai Canc Inst, State Key Lab Oncogenes & Related Genes, Shanghai, Peoples R China

Recommended Citation:
Han, Guanghui,Ye, Mingliang,Zhou, Houjiang,et al. Large-scale phosphoproteome analysis of human liver tissue by enrichment and fractionation of phosphopeptides with strong anion exchange chromatography[J]. PROTEOMICS,2008,8(7):1346-1361.
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