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Immobilized zirconium ion affinity chromatography for specific enrichment of phosphopeptides in phosphoproteome analysis
Feng, Shun; Ye, Mingliang; Zhou, Houjiang; Jiang, Xiaogang; Jiang, Xingning; Zou, Hanfa; Gong, Bolin; Zou HF(邹汉法); Zou HF(邹汉法)
刊名MOLECULAR & CELLULAR PROTEOMICS
2007-09-01
ISSN1535-9476
DOI10.1074/mcp.T600071-MCP200
6期:9页:1656-1665
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
关键词[WOS]TANDEM MASS-SPECTROMETRY ; PROTEIN-PHOSPHORYLATION ANALYSIS ; LARGE-SCALE ANALYSIS ; TYROSINE PHOSPHORYLATION ; SELECTIVE ENRICHMENT ; MONOLITHIC CAPILLARY ; YEAST PROTEOME ; MS/MS ANALYSIS ; LC-MS/MS ; IDENTIFICATION
英文摘要Large scale characterization of phosphoproteins requires highly specific methods for purification of phosphopeptides because of the low abundance of phosphoproteins and substoichiometry of phosphorylation. Enrichment of phosphopeptides from complex peptide mixtures by IMAC is a popular way to perform phosphoproteome analysis. However, conventional IMAC adsorbents with iminodiacetic acid as the chelating group to immobilize Fe3+ lack enough specificity for efficient phosphoproteome analysis. Here we report a novel IMAC adsorbent through Zr4+ chelation to the phosphonate- modified poly( glycidyl methacrylate- co- ethylene dimethacrylate) polymer beads. The high specificity of Zr4+- IMAC adsorbent was demonstrated by effectively enriching phosphopeptides from the digest mixture of phosphoprotein (alpha- or beta- casein) and bovine serum albumin with molar ratio at 1: 100. Zr(4 +)IMAC adsorbent was also successfully applied for the analysis of mouse liver phosphoproteome, resulting in the identification of 153 phosphopeptides ( 163 phosphorylation sites) from 133 proteins in mouse liver lysate. Significantly more phosphopeptides were identified than by the conventional Fe3+- IMAC approach, indicating the excellent performance of the Zr4 IMAC approach. The high specificity of Zr4+- IMAC adsorbent was found to mainly result from the strong interaction between chelating Zr4+ and phosphate group on phosphopeptides. Enrichment of phosphopeptides by Zr4 +- IMAC provides a powerful approach for large scale phosphoproteome analysis.
语种英语
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WOS记录号WOS:000249237200015
引用统计
被引频次:196[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/98729
专题中国科学院大连化学物理研究所
通讯作者Zou HF(邹汉法); Zou HF(邹汉法)
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog Res & Anal Ctr, Dalian 116023, Peoples R China
2.Xinjiang Univ, Coll Chem & Chem Engn, Urumqi 830046, Xinjiang, Peoples R China
3.Ningxia Univ, Key Lab Biotechnol, Yin Chuan 750021, Peoples R China
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GB/T 7714
Feng, Shun,Ye, Mingliang,Zhou, Houjiang,et al. Immobilized zirconium ion affinity chromatography for specific enrichment of phosphopeptides in phosphoproteome analysis[J]. MOLECULAR & CELLULAR PROTEOMICS,2007,6(9):1656-1665.
APA Feng, Shun.,Ye, Mingliang.,Zhou, Houjiang.,Jiang, Xiaogang.,Jiang, Xingning.,...&邹汉法.(2007).Immobilized zirconium ion affinity chromatography for specific enrichment of phosphopeptides in phosphoproteome analysis.MOLECULAR & CELLULAR PROTEOMICS,6(9),1656-1665.
MLA Feng, Shun,et al."Immobilized zirconium ion affinity chromatography for specific enrichment of phosphopeptides in phosphoproteome analysis".MOLECULAR & CELLULAR PROTEOMICS 6.9(2007):1656-1665.
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