DICP OpenIR
Subject Area物理化学
Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach
Song, Chunxia3; Wang, Fangjun1,3; Ye, Mingliang3; Cheng, Kai3; Chen, Rui3; Zhu, Jun3; Tan, Yexiong2; Wang, Hongyang2; Figeys, Daniel1; Zou, Hanfa3; Zou HF(邹汉法); DanielFigeys
Source PublicationANALYTICAL CHEMISTRY
2011-10-15
ISSN0003-2700
DOI10.1021/ac201299j
Volume83Issue:20Pages:7755-7762
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank1,1
WOS HeadingsScience & Technology ; Physical Sciences
WOS SubjectChemistry, Analytical
WOS Research AreaChemistry
WOS KeywordMASS-SPECTROMETRY ; QUANTITATIVE PROTEOMICS ; CELL-CULTURE ; AMINO-ACIDS ; MAP KINASE ; PHOSPHORYLATION ; SILAC ; EXPRESSION ; SEPARATION ; CANCER
AbstractImprovement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach; 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.
Language英语
WOS IDWOS:000295817500026
Citation statistics
Cited Times:43[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/115283
Collection中国科学院大连化学物理研究所
Corresponding AuthorZou HF(邹汉法); DanielFigeys
Affiliation1.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
GB/T 7714
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.
APA Song, Chunxia.,Wang, Fangjun.,Ye, Mingliang.,Cheng, Kai.,Chen, Rui.,...&DanielFigeys.(2011).Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach.ANALYTICAL CHEMISTRY,83(20),7755-7762.
MLA Song, Chunxia,et al."Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach".ANALYTICAL CHEMISTRY 83.20(2011):7755-7762.
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