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题名: Phosphoproteome analysis of human liver tissue by long-gradient nanoflow LC coupled with multiple stage MS analysis
作者: Han, Guanghui1;  Ye, Mingliang1;  Liu, Hongwei1;  Song, Chunxia1;  Sun, Deguang2;  Wu, Yifeng2;  Jiang, Xinning1;  Chen, Rui1;  Wang, Chunli1;  Wang, Liming2;  Zou, Hanfa1
通讯作者: 邹汉法
关键词: Fractionation ;  Human liver ;  Long-gradient elution ;  Phosphoproteome analysis
刊名: ELECTROPHORESIS
发表日期: 2010-03-01
DOI: 10.1002/elps.200900493
卷: 31, 期:6, 页:1080-1089
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 ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: Reversible protein phosphorylation plays a critical role in liver development and function Comprehensively cataloging the phosphoproteins and their phosphorylation sites in human liver tissue will facilitate the understanding of physiological and pathological mechanisms of liver Owing to lacking of efficient approach to fractionate phosphopeptides, nanoflow-RPLC with long-gradient elution was applied to reduce the complexity of the phosphopeptides in this study Two approaches were performed to further improve the coverage of phosphoproteome analysis of human liver tissue. In one approach, ten-replicated long-gradient LC-MS/MS runs were performed to analyze the enriched phosphopeptides, which resulted in the localization of 1080 phosphorylation sites from 495 proteins In another approach, proteins from liver tissue were first fractionated by SDS-PAGE and then long-gradient LC-MS/MS analysis was performed to analyze the phosphopeptides derived from each fraction, which resulted in the localization of 1786 phosphorylation sites from 911 proteins The two approaches showed the complementation in phosphoproteome analysis of human liver tissue. Combining the results of the two approaches, identification of 2225 nonredundant phosphorylation sites from 1023 proteins was obtained. The confidence of phosphopeptide identifications was strictly controlled with false discovery rate (FDR) <= 1% by a MS(2)/MS(3) target-decoy database search approach Among the localized 2225 phosphorylated sites, as many as 70.07% (1559 phosphorylated sites) were also reported by others, which confirmed the high confidence of the sites determined in this study. Considering the data acquired from low accuracy mass spectrometer and processed by a conservative MS(2)/MS(3) target-decoy approach, the number of localized phosphorylation sites obtained for human liver tissue in this study is quite impressive
关键词[WOS]: LARGE-SCALE ANALYSIS ;  MASS-SPECTROMETRY ;  PROTEIN-KINASE ;  MOUSE-LIVER ;  IN-VIVO ;  INDIVIDUAL PHOSPHOPROTEINS ;  PHOSPHORYLATION ANALYSIS ;  AFFINITY-CHROMATOGRAPHY ;  MONOLITHIC COLUMNS ;  SHOTGUN STRATEGY
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000276807300013
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/103057
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 R&A Ctr, Dalian 116023, Peoples R China
2.Dalian Med Univ, Affiliated Hosp 2, Dalian, Peoples R China

Recommended Citation:
Han, Guanghui,Ye, Mingliang,Liu, Hongwei,et al. Phosphoproteome analysis of human liver tissue by long-gradient nanoflow LC coupled with multiple stage MS analysis[J]. ELECTROPHORESIS,2010,31(6):1080-1089.
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