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Enrichment of phosphopeptides by Fe3+-immobilized mesoporous nanoparticles of MCM-41 for MALDI and nano-LC-MS/MS analysis
Pan, Chensong; Ye, Mingliang; Liu, Yuge; Feng, Shun; Jiang, Xiaogang; Han, Guanghui; Zhu, Junjie; Zou, Hanfa; Zou HF(邹汉法); Zou HF(邹汉法)
KeywordPhosphopeptides Maldi fe3+-immobilized Mcm-41 Nano-lc-ms/ms Enrichment
Source PublicationJOURNAL OF PROTEOME RESEARCH
2006-11-03
ISSN1535-3893
DOI10.1021/pr0600125
Volume5Issue:11Pages:3114-3124
Indexed BySCI
SubtypeArticle
Department18
Funding Project1809
Contribution Rank1;1
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectBiochemical Research Methods
WOS Research AreaBiochemistry & Molecular Biology
WOS KeywordFLIGHT-MASS-SPECTROMETRY ; ION AFFINITY-CHROMATOGRAPHY ; PROTEIN-PHOSPHORYLATION ; MOLECULAR-SIEVES ; PHOSPHOPROTEOME ANALYSIS ; MS ANALYSIS ; ADSORPTION ; SITES ; TIME ; IDENTIFICATION
AbstractFe3+-immobilized mesoporous molecular sieves MCM-41 with particle size of ca. 600 nm and pore size of ca. 3 nm is synthesized and applied to selectively trap and separate phosphopeptides from tryptic digest of proteins. For the capture of phosphopeptides, typically 10 mu L of tryptic digest solution was first diluted to 1 mL by solution of ACN/0.1% TFA (50:50, v/v) and incubated with 10 mu L of 0.1% acetic acid dispersed Fe3+-immobilized MCM-41 for 1 h under vibration. Fe3+-immobilized MCM-41 with trapped phosphopeptides was separated by centrifugation. The deposition was first washed with a volume of 300 mu L of solution containing 100 mM NaCl in ACN/0.1% TFA (50:50, v/v) and followed by a volume of 300 mu L of solution of 0.1% acetic acid to remove nonspecifically bound peptides. The nanoparticles with trapped phosphopeptides are mixed with 2,5-dihydroxybenzoic acid (2,5-DHB) and deposited onto the target for analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). It was found that phosphopeptides from tryptic digest of alpha-casein and beta-casein are effectively and specifically trapped on Fe3+-immobilized MCM-41 with few peptides nonspecifically adsorbed. After the extraction by Fe3+-immobilized MCM-41, the suppression to the detection of phosphopeptides caused by abundant nonphosphopeptides from tryptic digest is effectively eliminated, and the detection of phosphopeptides by MALDI is greatly enhanced with the value of signal-to-noise (S/N) increased by more than an order of magnitude. It is demonstrated that the mechanism of the adsorption of phosphopeptides on Fe3+-immobilized MCM-41 is based on the interaction between the Fe3+ and the phosphate group. Finally, Fe3+-immobilized MCM-41 is applied to extract phosphopeptides from tryptic digest of the lysate of mouse liver for phosphoproteome analysis by nano-LC-MS/MS.
Language英语
URL查看原文
WOS IDWOS:000241755400025
Citation statistics
Cited Times:63[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/97477
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&A Ctr, Dalian 116023, Peoples R China
2.Nanjing Univ, Dept Chem, Nanjing 210093, Peoples R China
Recommended Citation
GB/T 7714
Pan, Chensong,Ye, Mingliang,Liu, Yuge,et al. Enrichment of phosphopeptides by Fe3+-immobilized mesoporous nanoparticles of MCM-41 for MALDI and nano-LC-MS/MS analysis[J]. JOURNAL OF PROTEOME RESEARCH,2006,5(11):3114-3124.
APA Pan, Chensong.,Ye, Mingliang.,Liu, Yuge.,Feng, Shun.,Jiang, Xiaogang.,...&邹汉法.(2006).Enrichment of phosphopeptides by Fe3+-immobilized mesoporous nanoparticles of MCM-41 for MALDI and nano-LC-MS/MS analysis.JOURNAL OF PROTEOME RESEARCH,5(11),3114-3124.
MLA Pan, Chensong,et al."Enrichment of phosphopeptides by Fe3+-immobilized mesoporous nanoparticles of MCM-41 for MALDI and nano-LC-MS/MS analysis".JOURNAL OF PROTEOME RESEARCH 5.11(2006):3114-3124.
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