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学科主题: 物理化学
题名: Organic-inorganic hybrid silica monolith based immobilized titanium ion affinity chromatography column for analysis of mitochondrial phosphoproteome
作者: Hou CY(侯春彦) ;  Ma JF(马俊峰) ;  Tao DY(陶定银) ;  Liang Z(梁振) ;  Zhang LH(张丽华) ;  Zhang YK(张玉奎)
会议名称: 全球第九届人类蛋白质组学会议
会议日期: 2010-9-19
出版日期: 2010-09-19
会议地点: 澳大利亚
通讯作者: 张丽华
部门归属: 生物技术部
主办者: University of New South Wales
摘要: Phosphorylation is a reversible event affecting both the folding and function of proteins, regulating essential functions such as cell division, signal transduction and enzymatic activity. However, due to the dynamic nature of proteomes and low stoichiometry of phsophoproteins, the selective enrichment of phosphopetides is still indispensable prior to MS analysis. In our recent work, 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 μm i.d. capillary by sol-gel method with tetraethoxysilane and 3- aminopropyltriethoxysilane as precursors. Subsequently, amine groups were functionalized by glutaraldehyde, and then activated with (aminomethyl) phosphonic acid, followed by Ti4+ chelation. By such a monolithic Ti4+-IMAC column, 15 phosphopeptides could be effectively isolated in the digest mixture of -casein and BSA with the molar ratio as low as 1:200. With a synthetic phosphorylated peptide, YKVPQLEIVPNSpAEER, as the sample, the loading capacity and recovery of the Ti4+-IMAC monolithic column weremeasured to be 1.4 μmol/mL and 69% respectively. Such Ti4+-IMAC monolithic hybrid silica column was further applied to enrich phosphopeptides from mitochondrial enriched component from rat liver. Totally, 226 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 Ti4+-IMAC column might provide a promising tool for large-scale phosphopeptide enrichment, facilitating the in-depth understanding of the biological functions of phosphoproteomes.
语种: 中文
内容类型: 会议论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/114164
Appears in Collections:中国科学院大连化学物理研究所_会议论文

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Recommended Citation:
Hou CY,Ma JF,Tao DY,et al. Organic-inorganic hybrid silica monolith based immobilized titanium ion affinity chromatography column for analysis of mitochondrial phosphoproteome[C]. 见:全球第九届人类蛋白质组学会议. 澳大利亚. 2010-9-19.
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