DICP OpenIR
Subject Area物理化学
Depletion of Acidic Phosphopeptides by SAX To Improve the Coverage for the Detection of Basophilic Kinase Substrates
Dong, Mingming1,2; Ye, Mingliang1; Cheng, Kai1,2; Song, Chunxia1,2; Pan, Yanbo1,2; Wang, Chunli1,2; Bian, Yangyang1,2; Zou, Hanfa1; Ye ML(叶明亮); Zou HF(邹汉法)
KeywordPhosphoproteome Analysis Sax Depletion Of Acidic Phosphopeptides Basophilic Kinase Substrates
Source PublicationJOURNAL OF PROTEOME RESEARCH
2012-09-01
ISSN1535-3893
DOI10.1021/pr300503z
Volume11Issue:9Pages:4673-4681
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank1,1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordION AFFINITY-CHROMATOGRAPHY ; TITANIUM-DIOXIDE MICROCOLUMNS ; MASS-SPECTROMETRIC ANALYSIS ; PROTEIN-PHOSPHORYLATION ; PHOSPHOPROTEOME ANALYSIS ; SIGNALING NETWORKS ; IN-VIVO ; ENRICHMENT ; PEPTIDES ; IDENTIFICATION
AbstractThe 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.
Language英语
WOS IDWOS:000308389900025
Citation statistics
Cited Times:22[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/118241
Collection中国科学院大连化学物理研究所
Corresponding AuthorYe ML(叶明亮); Zou HF(邹汉法)
Affiliation1.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
GB/T 7714
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.
APA Dong, Mingming.,Ye, Mingliang.,Cheng, Kai.,Song, Chunxia.,Pan, Yanbo.,...&邹汉法.(2012).Depletion of Acidic Phosphopeptides by SAX To Improve the Coverage for the Detection of Basophilic Kinase Substrates.JOURNAL OF PROTEOME RESEARCH,11(9),4673-4681.
MLA Dong, Mingming,et al."Depletion of Acidic Phosphopeptides by SAX To Improve the Coverage for the Detection of Basophilic Kinase Substrates".JOURNAL OF PROTEOME RESEARCH 11.9(2012):4673-4681.
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