Determination of CK2 Specificity and Substrates by Proteome-Derived Peptide Libraries

	Determination of CK2 Specificity and Substrates by Proteome-Derived Peptide Libraries
	Wang, Chunli 1,2; Ye, Mingliang 1; Bian, Yangyang 1,2; Liu, Fangjie 1,2; Cheng, Kai 1,2; Dong, Mingming 1,2; Dong, Jing 1; Zou, Hanfa 1
刊名	journal of proteome research
	2013-08-01
卷号	12 期号:8 页码:3813-3821
关键词	peptide library casein kinase 2 specificity substrates quantitative phosphoproteomics
英文摘要	understanding the specificity of kinases enables prediction of their substrates and uncovering kinase functions in signaling pathways. traditionally synthesized peptide libraries are used to determine the kinase specificity. in this study, a proteomics-based method was developed to determine the specificity of kinase by taking the advantages of proteome-derived peptide libraries and quantitative proteomics. proteome-derived peptide libraries were constructed by digesting proteins in total cell lysate followed with dephosphorylation of the resulting peptides. after incubating the peptide libraries with/without ck2 for in vitro kinase assay, stable isotopic labeling based quantitative phosphoproteomics was applied to distinguish the in vitro phosphosites generated by ck2. by using the above approach, 404 ck2 in vitro phosphosites were identified by id lc-ms/ms. those sites allowed the statistic determination of the ck2 specificity. in addition to the easy construction of the proteome-derived peptide library, another significant advantage of this method over the method with synthesized peptide libraries is that the identified phosphosites could be directly mapped to proteins for the screening of putative kinase substrates. it was found that the confidence for substrate identification could be significantly improved by comparing the in vitro ck2 sites with the in vivo sites identified by phosphoproteomics analysis of the same cell lines. by applying this integrated strategy, 138 phosphosites from 105 putative ck2 substrates of high confidence were determined.
WOS标题词	science & technology ; life sciences & biomedicine
类目[WOS]	biochemical research methods
研究领域[WOS]	biochemistry & molecular biology
关键词[WOS]	ion affinity-chromatography ; kinase ck2 ; phosphoproteome analysis ; mass-spectrometry ; phosphorylation ; enrichment ; leukemia ; weblogo ; target
收录类别	SCI
语种	英语
WOS记录号	WOS:000322852800025
公开日期	2015-11-10
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/137868]
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.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Wang, Chunli,Ye, Mingliang,Bian, Yangyang,et al. Determination of CK2 Specificity and Substrates by Proteome-Derived Peptide Libraries[J]. journal of proteome research,2013,12(8):3813-3821.
APA	Wang, Chunli.,Ye, Mingliang.,Bian, Yangyang.,Liu, Fangjie.,Cheng, Kai.,...&Zou, Hanfa.(2013).Determination of CK2 Specificity and Substrates by Proteome-Derived Peptide Libraries.journal of proteome research,12(8),3813-3821.
MLA	Wang, Chunli,et al."Determination of CK2 Specificity and Substrates by Proteome-Derived Peptide Libraries".journal of proteome research 12.8(2013):3813-3821.