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Phosphoproteome analysis of human liver tissue by long-gradient nanoflow LC coupled with multiple stage MS analysis
Han, Guanghui1; Ye, Mingliang1; Liu, Hongwei1; Song, Chunxia1; Sun, Deguang2; Wu, Yifeng2; Jiang, Xinning1; Chen, Rui1; Wang, Chunli1; Wang, Liming2; Zou, Hanfa1; Zou HF(邹汉法)
关键词Fractionation Human Liver Long-gradient Elution Phosphoproteome Analysis
刊名ELECTROPHORESIS
2010-03-01
ISSN10.1002/elps.200900493
DOI10.1002/elps.200900493
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
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
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
语种英语
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WOS记录号WOS:000276807300013
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被引频次:47[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/321008/103057
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
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GB/T 7714
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.
APA Han, Guanghui.,Ye, Mingliang.,Liu, Hongwei.,Song, Chunxia.,Sun, Deguang.,...&邹汉法.(2010).Phosphoproteome analysis of human liver tissue by long-gradient nanoflow LC coupled with multiple stage MS analysis.ELECTROPHORESIS,31(6),1080-1089.
MLA Han, Guanghui,et al."Phosphoproteome analysis of human liver tissue by long-gradient nanoflow LC coupled with multiple stage MS analysis".ELECTROPHORESIS 31.6(2010):1080-1089.
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