Organic-Inorganic Hybrid Silica Monolith Based Immobilized Titanium Ion Affinity Chromatography Column for Analysis of Mitochondrial Phosphoproteome

	Organic-Inorganic Hybrid Silica Monolith Based Immobilized Titanium Ion Affinity Chromatography Column for Analysis of Mitochondrial Phosphoproteome
	Hou, Chunyan 1,2; Ma, Junfeng 1,2; Tao, Dingyin 1,2; Shan, Yichu 1; Liang, Zhen 1; Zhang, Lihua 1; Zhang, Yukui 1
刊名	journal of proteome research
	2010-08-01
卷号	9 期号:8 页码:4093-4101
关键词	immobilized metal ion affinity chromatography organic-inorganic hybrid silica monolith titanium ion phosphopeptide enrichment mitochondrial phosphoproteome
通讯作者	张丽华
产权排序	1;1
英文摘要	a novel kind of immobilized metal affinity chromatography (imac) column based on organic-inorganic hybrid silica monolith has been developed. the monolithic support was prepared in a 250 pm i.d. capillary by the sol-gel method with tetraethoxysilane (teos) and 3-aminopropyltriethoxysilane (aptes) as precursors. subsequently, amine groups were functionalized by glutaraldehyde, and then activated with (aminomethyl) phosphonic acid, followed by chelation. by such a hybrid silica monolithic ti(4+)-imac column, 15 phosphopeptides were effectively isolated from the digest mixture of cc-casein and bsa with the molar ratio as low as 1:200, illustrating its superior selectivity. with a synthetic phosphorylated peptide, ykvpqleivpnspaeer, as the sample, the loading capacity and recovery of the ti(4+)-imac monolithic column were measured to be 1.4 mu mol/ml and 69%, respectively. such an imac monolithic column was further applied to enrich phosphopeptides from rat liver mitochondria. in total, 224 unique phosphopeptides, corresponding to 148 phosphoprotein groups, were identified by duplicate nanorplc-ltq ms/ms/ms runs with a false-positive rate of less than 1% at the peptide level. these results demonstrate that the hybrid silica monolith based ti(4+)-imac column might provide a promising tool for large-scale phosphopeptide enrichment, facilitating the in-depth understanding of the biological functions of phosphoproteomes.
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]	protein-phosphorylation ; mass-spectrometry ; phosphopeptide enrichment ; in-vivo ; identification ; efficiency ; erk1/2
收录类别	SCI
原文出处	false
语种	英语
WOS记录号	WOS:000280583700031
公开日期	2010-11-30
内容类型	期刊论文
源URL	[http://159.226.238.44/handle/321008/103563]
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 Res & Anal Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
推荐引用方式 GB/T 7714	Hou, Chunyan,Ma, Junfeng,Tao, Dingyin,et al. Organic-Inorganic Hybrid Silica Monolith Based Immobilized Titanium Ion Affinity Chromatography Column for Analysis of Mitochondrial Phosphoproteome[J]. journal of proteome research,2010,9(8):4093-4101.
APA	Hou, Chunyan.,Ma, Junfeng.,Tao, Dingyin.,Shan, Yichu.,Liang, Zhen.,...&Zhang, Yukui.(2010).Organic-Inorganic Hybrid Silica Monolith Based Immobilized Titanium Ion Affinity Chromatography Column for Analysis of Mitochondrial Phosphoproteome.journal of proteome research,9(8),4093-4101.
MLA	Hou, Chunyan,et al."Organic-Inorganic Hybrid Silica Monolith Based Immobilized Titanium Ion Affinity Chromatography Column for Analysis of Mitochondrial Phosphoproteome".journal of proteome research 9.8(2010):4093-4101.