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题名: Organic-Inorganic Hybrid Silica Monolith Based Immobilized Titanium Ion Affinity Chromatography Column for Analysis of Mitochondrial Phosphoproteome
作者: Hou, Chunyan1, 2;  Ma, Junfeng1, 2;  Tao, Dingyin1, 2;  Shan, Yichu1;  Liang, Zhen1;  Zhang, Lihua1;  Zhang, Yukui1
通讯作者: 张丽华
关键词: immobilized metal ion affinity chromatography ;  organic-inorganic hybrid silica monolith ;  titanium ion ;  phosphopeptide enrichment ;  mitochondrial phosphoproteome
刊名: JOURNAL OF PROTEOME RESEARCH
发表日期: 2010-08-01
DOI: 10.1021/pr100294z
卷: 9, 期:8, 页:4093-4101
收录类别: SCI
文章类型: Article
部门归属: 18
项目归属: 1810
产权排名: 1;1
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine
类目[WOS]: Biochemical Research Methods
研究领域[WOS]: Biochemistry & Molecular Biology
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.
关键词[WOS]: PROTEIN-PHOSPHORYLATION ;  MASS-SPECTROMETRY ;  PHOSPHOPEPTIDE ENRICHMENT ;  IN-VIVO ;  IDENTIFICATION ;  EFFICIENCY ;  ERK1/2
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000280583700031
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/103563
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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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

Recommended Citation:
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.
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