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学科主题: 物理化学
题名: Depletion of Acidic Phosphopeptides by SAX To Improve the Coverage for the Detection of Basophilic Kinase Substrates
作者: Dong, Mingming1, 2;  Ye, Mingliang1;  Cheng, Kai1, 2;  Song, Chunxia1, 2;  Pan, Yanbo1, 2;  Wang, Chunli1, 2;  Bian, Yangyang1, 2;  Zou, Hanfa1
通讯作者: 叶明亮 ;  邹汉法
关键词: phosphoproteome analysis ;  SAX ;  depletion of acidic phosphopeptides ;  basophilic kinase substrates
刊名: JOURNAL OF PROTEOME RESEARCH
发表日期: 2012-09-01
DOI: 10.1021/pr300503z
卷: 11, 期:9, 页:4673-4681
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1809
产权排名: 1,1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: The Ser/Thr protein kinases fall into three major subgroups, pro-directed, basophilic, and acidophilic, on the basis of the types of substrate sequences that they preferred. Despite many phosphoproteomics efforts that have been taken for global profiling of phosphopeptides, methodologies focusing on analyzing a particular type of kinase substrates have seldom been reported. Selective enrichment of phosphopeptides from basophilic kinase substrates is difficult because basophilic motifs are cleaved by trypsin during digestion. In this study, we develop a negative enrichment strategy to enhance the identification of basophilic kinase substrates. This method is based on an observation that high pH strong anion exchange (SAX) chromatography can separate tryptic phosphopeptides according to the number of acidic amino acidic residues that they have. Thus, SAX was applied to deplete acidic phosphopeptides from the phosphopeptide mixture, which improved the coverage for the detection of basophilic kinase substrates. The SAX depletion approach was further combined with online SCX-RP separation for large-scale analysis of mouse liver phosphoproteome, which resulted in the identification of 6944 phosphorylated sites. It was found that motifs associated with basophilic kinases prevail for these identified phosphorylated sites.
关键词[WOS]: ION AFFINITY-CHROMATOGRAPHY ;  TITANIUM-DIOXIDE MICROCOLUMNS ;  MASS-SPECTROMETRIC ANALYSIS ;  PROTEIN-PHOSPHORYLATION ;  PHOSPHOPROTEOME ANALYSIS ;  SIGNALING NETWORKS ;  IN-VIVO ;  ENRICHMENT ;  PEPTIDES ;  IDENTIFICATION
语种: 英语
WOS记录号: WOS:000308389900025
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/118241
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 Res & Anal Ctr, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China

Recommended Citation:
Dong, Mingming,Ye, Mingliang,Cheng, Kai,et al. Depletion of Acidic Phosphopeptides by SAX To Improve the Coverage for the Detection of Basophilic Kinase Substrates[J]. JOURNAL OF PROTEOME RESEARCH,2012,11(9):4673-4681.
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