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Specific phosphopeptide enrichment with immobilized titanium ion affinity chromatography adsorbent for phosphoproteome analysis
Zhou, Houjiang; Ye, Mingliang; Dong, Jing; Han, Guanghui; Jiang, Xinning; Wu, Renan; Zou, Hanfa; Zou HF(邹汉法); Zou HF(邹汉法)
KeywordImmobilized Metal Ion Affinity Chromatography Titanium Ion Phosphate Polymer Phosphopeptide Enrichment Phosphoproteome Analysis
Source PublicationJOURNAL OF PROTEOME RESEARCH
2008-09-01
ISSN1535-3893
DOI10.1021/pr800223m
Volume7Issue:9Pages:3957-3967
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 KeywordTANDEM MASS-SPECTROMETRY ; SCALE PHOSPHORYLATION ANALYSIS ; HIGHLY SELECTIVE ENRICHMENT ; NANO-LC-MS/MS ; SACCHAROMYCES-CEREVISIAE ; MAGNETIC NANOPARTICLES ; METHYL ESTERIFICATION ; PEPTIDE MIXTURES ; MOUSE-LIVER ; MS ANALYSIS
AbstractThe elucidation of protein post-translational modifications, such as phosphorylation, remains a challenging analytical task for proteomic studies. Since many of the proteins targeted for phosphorylation are low in abundance and phosphorylation is typically substoichiometric, a prerequisite for their identification is the specific enrichment of phosphopeptide prior to mass spectrometric analysis. Here, we presented a new method termed as immobilized titanium ion affinity chromatography (Ti4+-IMAC) for enriching phosphopeptides. A phosphate polymer, which was prepared by direct polymerization of monomers containing phosphate groups, was applied to immobilize Ti4+ through the chelating interaction between phosphate groups on the polymer and Ti4+. The resulting Ti4+-IMAC resin specifically isolates phosphopeptides from a digest mixture of standard phosphoproteins and nonphosphoprotein (BSA) in a ratio as low as 1:500. Ti4+-IMAC was further applied for phosphoproteome analysis of mouse liver. We also compared Ti4+-IMAC to other enrichment methods including Fe3+-IMAC, Zr4+-IMAC, TiO2 and ZrCl2 and demonstrate superior selectivity and efficiency of Ti4+-IMAC for the isolation and enrichment of phosphopeptides. The high specificity and efficiency of phosphopeptide enrichment by Ti4+-IMAC mainly resulted from the flexibility of immobilized titanium ion with spacer arm linked to polymer beads as well as the specific interaction between immobilized titanium ion and phosphate group on phosphopeptides.
Language英语
URL查看原文
WOS IDWOS:000259015500031
Citation statistics
Cited Times:194[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/100199
Collection中国科学院大连化学物理研究所
Corresponding AuthorZou HF(邹汉法); Zou HF(邹汉法)
AffiliationChinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Dalian 116023, Peoples R China
Recommended Citation
GB/T 7714
Zhou, Houjiang,Ye, Mingliang,Dong, Jing,et al. Specific phosphopeptide enrichment with immobilized titanium ion affinity chromatography adsorbent for phosphoproteome analysis[J]. JOURNAL OF PROTEOME RESEARCH,2008,7(9):3957-3967.
APA Zhou, Houjiang.,Ye, Mingliang.,Dong, Jing.,Han, Guanghui.,Jiang, Xinning.,...&邹汉法.(2008).Specific phosphopeptide enrichment with immobilized titanium ion affinity chromatography adsorbent for phosphoproteome analysis.JOURNAL OF PROTEOME RESEARCH,7(9),3957-3967.
MLA Zhou, Houjiang,et al."Specific phosphopeptide enrichment with immobilized titanium ion affinity chromatography adsorbent for phosphoproteome analysis".JOURNAL OF PROTEOME RESEARCH 7.9(2008):3957-3967.
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