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题名: Specific phosphopeptide enrichment with immobilized titanium ion affinity chromatography adsorbent for phosphoproteome analysis
作者: Zhou, Houjiang1;  Ye, Mingliang1;  Dong, Jing1;  Han, Guanghui1;  Jiang, Xinning1;  Wu, Renan1;  Zou, Hanfa1
通讯作者: 邹汉法
关键词: immobilized metal ion affinity chromatography ;  titanium ion ;  phosphate polymer ;  phosphopeptide enrichment ;  phosphoproteome analysis
刊名: JOURNAL OF PROTEOME RESEARCH
发表日期: 2008-09-01
DOI: 10.1021/pr800223m
卷: 7, 期:9, 页:3957-3967
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 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.
关键词[WOS]: TANDEM MASS-SPECTROMETRY ;  SCALE PHOSPHORYLATION ANALYSIS ;  HIGHLY SELECTIVE ENRICHMENT ;  NANO-LC-MS/MS ;  SACCHAROMYCES-CEREVISIAE ;  MAGNETIC NANOPARTICLES ;  METHYL ESTERIFICATION ;  PEPTIDE MIXTURES ;  MOUSE-LIVER ;  MS ANALYSIS
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000259015500031
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/100199
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 R&A Ctr, Dalian 116023, Peoples R China

Recommended Citation:
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.
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