Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach

	Improvement of the Quantification Accuracy and Throughput for Phosphoproteome Analysis by a Pseudo Triplex Stable Isotope Dimethyl Labeling Approach
	Song, Chunxia 3; Wang, Fangjun 1,3; Ye, Mingliang 3; Cheng, Kai 3; Chen, Rui 3; Zhu, Jun 3; Tan, Yexiong 2; Wang, Hongyang 2; Figeys, Daniel 1; Zou, Hanfa 3
刊名	analytical chemistry
	2011-10-15
卷号	83 期号:20 页码:7755-7762
ISSN号	0003-2700
通讯作者	邹汉法 ; danielfigeys
产权排序	1,1
中文摘要	improvement of the quantification accuracy and throughput for phosphoproteome analysis by a pseudo triplex stable isotope dimethyl labeling approach
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.
学科主题	物理化学
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, analytical
研究领域[WOS]	chemistry
关键词[WOS]	mass-spectrometry ; quantitative proteomics ; cell-culture ; amino-acids ; map kinase ; phosphorylation ; silac ; expression ; separation ; cancer
收录类别	SCI
语种	英语
WOS记录号	WOS:000295817500026
公开日期	2012-07-09
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/115283]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
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
推荐引用方式 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.,...&Zou, Hanfa.(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.