Enrichment of phosphopeptides by Fe3+-immobilized mesoporous nanoparticles of MCM-41 for MALDI and nano-LC-MS/MS analysis

	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
刊名	journal of proteome research
	2006-11-03
卷号	5 期号:11 页码:3114-3124
关键词	phosphopeptides MALDI Fe3+-immobilized MCM-41 nano-LC-MS/MS enrichment
ISSN号	1535-3893
通讯作者	邹汉法
产权排序	1;1
英文摘要	fe3+-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.
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]	flight-mass-spectrometry ; ion affinity-chromatography ; protein-phosphorylation ; molecular-sieves ; phosphoproteome analysis ; ms analysis ; adsorption ; sites ; time ; identification
收录类别	SCI
原文出处	true
语种	英语
WOS记录号	WOS:000241755400025
公开日期	2010-11-30
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/97477]
专题	大连化学物理研究所_中国科学院大连化学物理研究所
作者单位	1.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
推荐引用方式 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.,...&Zou, Hanfa.(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.