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题名: Fe3+ immobilized metal affinity chromatography with silica monolithic capillary column for phosphoproteome analysis
作者: Feng, Shun;  Pan, Chensong;  Jiang, Xiaogang;  Xu, Songyu;  Zhou, Houjiang;  Ye, Mingliang;  Zou, Hanfa
通讯作者: 邹汉法
关键词: immobilized metal affinity chromatography ;  miniaturization ;  phosphoproteome analysis ;  silica monolithic column
刊名: PROTEOMICS
发表日期: 2007-02-01
DOI: 10.1002/pmic.200600045
卷: 7, 期:3, 页:351-360
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 ;  Biochemistry & Molecular Biology
研究领域[WOS]: Biochemistry & Molecular Biology
英文摘要: Immobilized metal affinity chromatography (IMAC) is a commonly used technique for phosphoproteome analysis due to its high affinity for adsorption of phosphopeptides. Miniaturization of IMAC column is essential for the analysis of a small amount of sample. Nanoscale IMAC column vas prepared by chemical m6dification of silica monolith with iminodiacetic acid (IDA) followed by the immobilization of Fe3+ ion inside the capillary. it was demonstrated that Fe3+-IDA silica monolithic IMAC capillary column could specifically capture the phosphopeptides from tryptic digest of (x-casein with analysis by MALDI-TOF MS. The silica monolithic IMAC capillary column was manually coupled with nanoflow RPLC/nanospray ESI mass spectrometer (mu RPLC-nanoESI MS) for phosphoproteome analysis. The system was validated by analysis of standard phosphoproteins and then it was applied to the analysis of protein phosphorylation in mouse liver lysate. Besides MS/MS spectra, MS/MS/MS spectra were also collected for neutral loss peak. After database search and manual validation with conservative criteria, 29 singly phosphorylated peptides were identified by analyzing a tryptic digest of only 12 mu g mouse liver lysate. The results demonstrated that the silica monolithic IMAC capillary column coupled with gRPLC-nanoESI MS was very suitable for the phosphoproteome analysis of minute sample.
关键词[WOS]: IONIZATION MASS-SPECTROMETRY ;  PROTEIN-PHOSPHORYLATION SITES ;  ION AFFINITY ;  LIQUID-CHROMATOGRAPHY ;  PROTEOMIC ANALYSIS ;  PHASE-SEPARATION ;  MALDI-MS ;  IDENTIFICATION ;  ONLINE ;  PHOSPHOPEPTIDES
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000244271500002
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/98459
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&D Ctr, Dalian 116023, Peoples R China
2.Xinjiang Univ, Coll Chem & Chem Engn, Urumqi, Xinjiang, Peoples R China

Recommended Citation:
Feng, Shun,Pan, Chensong,Jiang, Xiaogang,et al. Fe3+ immobilized metal affinity chromatography with silica monolithic capillary column for phosphoproteome analysis[J]. PROTEOMICS,2007,7(3):351-360.
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