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题名: Specific mixing facilitates the comparative quantification of phosphorylation sites with significant dysregulations
作者: Liu, Jing1, 2;  Xu, Bo1;  Liu, Zheyi1, 2;  Dong, Mingming1, 2;  Mao, Jiawei1, 2;  Zhou, Ye1, 2;  Chen, Jin1, 2;  Wang, Fangjun1;  Zou, Hanfa1
关键词: Mass spectrometry ;  Comparative phosphoproteome quantification ;  Stable isotope labeling ;  Mixing at specific ratio ;  Extreme relative abundance
刊名: ANALYTICA CHIMICA ACTA
发表日期: 2017-01-15
DOI: 10.1016/j.aca.2016.10.044
卷: 950, 页:129-137
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ;  Physical Sciences
类目[WOS]: Chemistry, Analytical
研究领域[WOS]: Chemistry
英文摘要: Mass spectrometry (MS) based quantitative analyses of proteome and proteome post-translational modifications (PTMs) play more and more important roles in biological, pharmaceutical and clinical studies. However, it is still a big challenge to accurately quantify the proteins or proteins PTM sites with extreme relative abundances in comparative protein samples, such as the significantly dysregulated ones. Herein, a novel quantification strategy, Mixing at Specific Ratio (MaSR) before isotope labeling, had been developed to improve the quantification accuracy and coverage of extreme proteins and protein phosphorylation sites. Briefly, the comparative protein samples were firstly mixed together at specific ratios of 9:1 and 1:9 (w/w), followed with mass differentiate light and heavy isotope labeling, respectively. The extreme proteins and protein phosphorylation sites, even if the newly expressed or disappeared ones, could be accurately quantified due to all of the proteins' relative abundances had been adjusted to 2 orders of magnitude (1/9-9) by this strategy. The number of quantified phosphorylation sites with more than 20 folds changes was improved about 10 times in comparative quantification of pervanadate stimulated phosphoproteome of HeLa cells, and 134 newly generated and 21 disappeared phosphorylation sites were solely quantified by the MaSR strategy. The significantly up-regulated phosphorylation sites were mainly involved in the key phosphoproteins regulating the insulin-related pathways, such as PI3K-AKT and RAS-MAPK pathways. Therefore, the MaSR strategy exhibits as a promising way in elucidating the biological processes with significant dysregulations. (C) 2016 Elsevier B.V. All rights reserved.
关键词[WOS]: GROWTH-FACTOR RECEPTOR ;  QUANTITATIVE PROTEOMICS ;  MASS-SPECTROMETRY ;  IN-VIVO ;  SILAC ;  MECHANISMS ;  EXPRESSION ;  THROUGHPUT ;  VANADATE ;  PROTEINS
语种: 英语
WOS记录号: WOS:000390629500014
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/151834
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.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China

Recommended Citation:
Liu, Jing,Xu, Bo,Liu, Zheyi,et al. Specific mixing facilitates the comparative quantification of phosphorylation sites with significant dysregulations[J]. ANALYTICA CHIMICA ACTA,2017,950:129-137.
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