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Fe3+ immobilized metal affinity chromatography with silica monolithic capillary column for phosphoproteome analysis
Feng, Shun; Pan, Chensong; Jiang, Xiaogang; Xu, Songyu; Zhou, Houjiang; Ye, Mingliang; Zou, Hanfa; Zou HF(邹汉法); Zou HF(邹汉法)
关键词Immobilized Metal Affinity Chromatography Miniaturization Phosphoproteome Analysis Silica Monolithic Column
刊名PROTEOMICS
2007-02-01
ISSN1615-9853
DOI10.1002/pmic.200600045
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
关键词[WOS]IONIZATION MASS-SPECTROMETRY ; PROTEIN-PHOSPHORYLATION SITES ; ION AFFINITY ; LIQUID-CHROMATOGRAPHY ; PROTEOMIC ANALYSIS ; PHASE-SEPARATION ; MALDI-MS ; IDENTIFICATION ; ONLINE ; PHOSPHOPEPTIDES
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.
语种英语
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WOS记录号WOS:000244271500002
引用统计
文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/98459
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 R&D Ctr, Dalian 116023, Peoples R China
2.Xinjiang Univ, Coll Chem & Chem Engn, Urumqi, Xinjiang, Peoples R China
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GB/T 7714
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.
APA Feng, Shun.,Pan, Chensong.,Jiang, Xiaogang.,Xu, Songyu.,Zhou, Houjiang.,...&邹汉法.(2007).Fe3+ immobilized metal affinity chromatography with silica monolithic capillary column for phosphoproteome analysis.PROTEOMICS,7(3),351-360.
MLA Feng, Shun,et al."Fe3+ immobilized metal affinity chromatography with silica monolithic capillary column for phosphoproteome analysis".PROTEOMICS 7.3(2007):351-360.
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