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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)
KeywordImmobilized Metal Affinity Chromatography Miniaturization Phosphoproteome Analysis Silica Monolithic Column
Source PublicationPROTEOMICS
2007-02-01
ISSN1615-9853
DOI10.1002/pmic.200600045
Volume7Issue:3Pages:351-360
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank1;1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordIONIZATION MASS-SPECTROMETRY ; PROTEIN-PHOSPHORYLATION SITES ; ION AFFINITY ; LIQUID-CHROMATOGRAPHY ; PROTEOMIC ANALYSIS ; PHASE-SEPARATION ; MALDI-MS ; IDENTIFICATION ; ONLINE ; PHOSPHOPEPTIDES
AbstractImmobilized 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.
Language英语
URL查看原文
WOS IDWOS:000244271500002
Citation statistics
Cited Times:69[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/98459
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)
Affiliation1.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
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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