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学科主题物理化学
In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis
Huang, Junfeng1,2; Qin, Hongqiang1; Dong, Jing1; Song, Chunxia1; Bian, Yangyang1,2; Dong, Mingming1,2; Cheng, Kai1,2; Wang, Fangjun1; Sun, Deguang3; Wang, Liming3; Ye, Mingliang1; Zou, Hanfa1; Ye ML(叶明亮); Zou HF(邹汉法)
关键词In Situ Sample processIng Approach Phosphoprotein Enrichment On-beads Digestion Dimethyl Labeling Solid Phase Labeling Phosphoproteome Quantification Human Liver Hepatocellular Carcinoma
刊名JOURNAL OF PROTEOME RESEARCH
2014-09-01
DOI10.1021/pr500454g
13期:9页:3896-3904
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Biochemical Research Methods
研究领域[WOS]Biochemistry & Molecular Biology
关键词[WOS]ION AFFINITY-CHROMATOGRAPHY ; PHOSPHORYLATED PROTEINS ; MASS-SPECTROMETRY ; SELECTIVE ENRICHMENT ; PHOSPHOPEPTIDES ; QUANTIFICATION ; LIVER ; IDENTIFICATION ; ADSORBENT ; PEPTIDES
英文摘要Current sample preparation protocols for quantitative phosphoproteome analysis are tedious and time-consuming. Here, a facile in situ sample processing approach (iSPA) is developed by using macroporous Ti(IV)-IMAC microspheres as the preparation "beds", where all sample preparation procedures including the enrichment of phosphoproteins, tryptic digestion of proteins, enrichment, and isotope labeling of phosphopeptides are performed in situ sequentially. As a result of the in situ processing design and the seamless procedures, extra steps for desalting and buffer exchanging, which are always required in conventional approaches, are avoided, and the sample loss and contamination could be greatly reduced. Thus, better sensitivity and accuracy for the quantitative phosphoproteome analysis were obtained. This strategy was further applied to differential phosphoproteome analysis of human liver tissues with or without hepatocellular carcinoma (HCC). In total, 8548 phosphorylation sites were confidently quantified from three replicate analyses of 0.5 mg of human liver protein extracts.
语种英语
WOS记录号WOS:000341345000004
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被引频次:6[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/321008/144524
专题中国科学院大连化学物理研究所
通讯作者Ye ML(叶明亮); Zou HF(邹汉法)
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
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
3.Dalian Med Univ, Affiliated Hosp 2, Dalian 116027, Peoples R China
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Huang, Junfeng,Qin, Hongqiang,Dong, Jing,et al. In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis[J]. JOURNAL OF PROTEOME RESEARCH,2014,13(9):3896-3904.
APA Huang, Junfeng.,Qin, Hongqiang.,Dong, Jing.,Song, Chunxia.,Bian, Yangyang.,...&邹汉法.(2014).In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis.JOURNAL OF PROTEOME RESEARCH,13(9),3896-3904.
MLA Huang, Junfeng,et al."In Situ Sample Processing Approach (iSPA) for Comprehensive Quantitative Phosphoproteome Analysis".JOURNAL OF PROTEOME RESEARCH 13.9(2014):3896-3904.
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