Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples

	Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples
	Wang, Fangjun; Han, Guanghui; Yu, Zhiyuan; Jiang, Xinning; Sun, Shutao; Chen, Rui; Ye, Mingliang; Zou, Hanfa
刊名	journal of separation science
	2010-07-01
卷号	33 期号:13 页码:1879-1887
关键词	Fractionating Human liver Phosphoproteome Strong anion-exchange Trap column
ISSN号	1615-9306
通讯作者	邹汉法
产权排序	1;1
英文摘要	it is one of the key issues to develop powerful fractionating method to increase the identification of the low-abundance phosphopeptides. in this study, a semi-online 2-d lc separation strategy based on three-step fractionation of the enriched peptides on strong anion-exchange trap column was developed. it was demonstrated that the sensitivity and phosphoproteome coverage obtained by this fractionating method with strong anion-exchange trap column is much higher than those by the conventional methods based on c18 trap column. in addition, when the same amount of sample was loaded, the number of identified phosphopeptides had increased 108%. combination of this three-step fractionation method with rplc-ms/ms analysis by 300 min rp-gradient separation was applied to phosphoproteome analysis of human liver proteins, and 853 unique phosphopeptides was positively identified from 500 mu g tryptic digest of human liver proteins. after three cycles' consecutive analyses, 1554 unique phosphopeptides and 1566 phosphorylated sites were totally identified from 735 phosphorylated proteins at a false discovery rate of < 1% in about 54h of analysis time.
WOS标题词	science & technology ; physical sciences
类目[WOS]	chemistry, analytical
研究领域[WOS]	chemistry
关键词[WOS]	spectrometry-based proteomics ; ion affinity-chromatography ; lc-ms/ms analysis ; phosphorylated peptides ; mass-spectrometry ; posttranslational modifications ; magnetic nanoparticles ; liquid-chromatography ; shotgun proteomics ; mouse-liver
收录类别	SCI
原文出处	false
语种	英语
WOS记录号	WOS:000280163900001
公开日期	2010-11-30
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/103147]
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	Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
推荐引用方式 GB/T 7714	Wang, Fangjun,Han, Guanghui,Yu, Zhiyuan,et al. Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples[J]. journal of separation science,2010,33(13):1879-1887.
APA	Wang, Fangjun.,Han, Guanghui.,Yu, Zhiyuan.,Jiang, Xinning.,Sun, Shutao.,...&Zou, Hanfa.(2010).Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples.journal of separation science,33(13),1879-1887.
MLA	Wang, Fangjun,et al."Fractionation of phosphopeptides on strong anion-exchange capillary trap column for large-scale phosphoproteome analysis of microgram samples".journal of separation science 33.13(2010):1879-1887.