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学科主题: 物理化学
题名: Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach
作者: Song, Chunxia3;  Wang, Fangjun1, 3;  Ye, Mingliang3;  Cheng, Kai3;  Chen, Rui3;  Zhu, Jun3;  Tan, Yexiong2;  Wang, Hongyang2;  Figeys, Daniel1;  Zou, Hanfa3
通讯作者: 邹汉法 ;  DanielFigeys
刊名: ANALYTICAL CHEMISTRY
发表日期: 2011-10-15
DOI: 10.1021/ac201299j
卷: 83, 期:20, 页:7755-7762
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
摘要: Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach
研究领域[WOS]: Chemistry
英文摘要: Accurately quantifying the changes of phosphorylation level on specific sites is crucial to understand the role of protein phosphorylation in physiological and pathological processes. Here, a pseudo triplex stable isotope dimethyl labeling approach was developed to improve the accuracy and the throughput of comprehensive quantitative phosphoproteome analyses. In this strategy, two identical samples are labeled with light and heavy isotopes, respectively, while another comparative sample is labeled with an intermediate isotope. Two replicated quantification results were achieved in just one experiment, and the relative standard deviation (RSD) criterion was used to control the quantification accuracy. Compared with the conventional duplex labeling approach, the number of quantified phosphopeptides increased nearly 50% and the experimental time was reduced by 50% under the same quantification accuracy. Combined with the automated online reversed phase-strong cation exchange-reversed phase (RP-SCX-RP) multidimensional separation system, a comparative phosphoproteome analysis of hepatocellular carcinoma (HCC) and normal human liver tissues was performed. Over 1800 phosphopeptides corresponding to similar to 2000 phosphorylation sites were quantified reliably in a 42 h multidimensional analysis. The pro-directed motifs, which were mainly associated with the extracellular signal-regulated kinases (ERKs), were observed as being overrepresented in the regulated phosphorylation sites, and some quantification results of phosphorylation sites were validated by the other studies. Therefore, this pseudo triplex labeling approach was demonstrated as a promising alternative for the comprehensive quantitative phosphoproteome analysis.
关键词[WOS]: MASS-SPECTROMETRY ;  QUANTITATIVE PROTEOMICS ;  CELL-CULTURE ;  AMINO-ACIDS ;  MAP KINASE ;  PHOSPHORYLATION ;  SILAC ;  EXPRESSION ;  SEPARATION ;  CANCER
语种: 英语
WOS记录号: WOS:000295817500026
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/115283
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.Univ Ottawa, Fac Med, Ottawa Inst Syst Biol, Dept Biochem Microbiol & Immunol, Ottawa, ON K1H 8M5, Canada
2.Second Mil Med Univ, Int Cooperat Lab Signal Transduct, Eastern Hepatobiliary Surg Inst, Shanghai 200438, Peoples R China
3.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China

Recommended Citation:
Song, Chunxia,Wang, Fangjun,Ye, Mingliang,et al. Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach[J]. ANALYTICAL CHEMISTRY,2011,83(20):7755-7762.
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