Comprehensive and Reliable Phosphorylation Site Mapping of Individual Phosphoproteins by Combination of Multiple Stage Mass Spectrometric Analysis with a Target-Decoy Database Search

	Comprehensive and Reliable Phosphorylation Site Mapping of Individual Phosphoproteins by Combination of Multiple Stage Mass Spectrometric Analysis with a Target-Decoy Database Search
	Han, Guanghui 1; Ye, Mingliang 1; Jiang, Xinning 1; Chen, Rui 1; Ren, Jian 2,3; Xue, Yu 2,3; Wang, Fangjun 1; Song, Chunxia 1; Yao, Xuebiao 2,3; Zou, Hanfa 1
刊名	analytical chemistry
	2009-07-15
卷号	81 期号:14 页码:5794-5805
通讯作者	邹汉法
产权排序	1;1
英文摘要	since the emergence of proteomics, much attention has been paid to the development of new technologies for phosphoproteomcis analysis. compared with large scale phosphorylation analysis at the proteome level, comprehensive and reliable phosphorylation site mapping of individual phosphoprotein is equally important. here, we present a modified target-decoy database search strategy for confident phosphorylation site analysis of individual phosphoproteins without manual interpretation of spectra. instead of using all protein sequences in a proteome database of an organism for the construction of a target-decoy database for phosphoproteome analysis, the composite database constructed for phosphorylation site analysis of individual phosphoproteins only included the sequences of the individual target proteins and a decoy version of a small inhomogeneous protein database. it was found that the confidence of phosphopeptide identifications could be effectively controlled when the acquired ms(2) and ms(3) spectra were searched against the above composite database followed with data processing. because of the small size of the composite database, the computation time for the database search is very short, which allows the adoption of low-specificity proteases for protein digestion to increase the coverage of phosphorylation site mapping. the sensitivity and comprehensive phosphorylation site mapping of this approach was demonstrated by using two standard phosphoprotein samples of alpha-casein and beta-casein, and this approach was further applied to analyze the phosphorylation of the cyclic amp-dependent protein kinase (pka), which resulted in the identification of 17 phosphorylation sites, including five novel sites on four pka subunits.
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, analytical
研究领域[WOS]	chemistry
关键词[WOS]	protein-phosphorylation ; in-vivo ; phosphopeptide identifications ; affinity-chromatography ; automatic validation ; signaling networks ; enrichment ; proteomics ; strategy ; fragmentation
收录类别	SCI
原文出处	false
语种	英语
WOS记录号	WOS:000268135000028
公开日期	2010-11-30
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/102971]
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.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China 2.Univ Sci & Technol China, Hefei Natl Lab Phys Sci Microscale, Hefei 230027, Peoples R China 3.Univ Sci & Technol China, Sch Life Sci, Hefei 230027, Peoples R China
推荐引用方式 GB/T 7714	Han, Guanghui,Ye, Mingliang,Jiang, Xinning,et al. Comprehensive and Reliable Phosphorylation Site Mapping of Individual Phosphoproteins by Combination of Multiple Stage Mass Spectrometric Analysis with a Target-Decoy Database Search[J]. analytical chemistry,2009,81(14):5794-5805.
APA	Han, Guanghui.,Ye, Mingliang.,Jiang, Xinning.,Chen, Rui.,Ren, Jian.,...&Zou, Hanfa.(2009).Comprehensive and Reliable Phosphorylation Site Mapping of Individual Phosphoproteins by Combination of Multiple Stage Mass Spectrometric Analysis with a Target-Decoy Database Search.analytical chemistry,81(14),5794-5805.
MLA	Han, Guanghui,et al."Comprehensive and Reliable Phosphorylation Site Mapping of Individual Phosphoproteins by Combination of Multiple Stage Mass Spectrometric Analysis with a Target-Decoy Database Search".analytical chemistry 81.14(2009):5794-5805.