DICP OpenIR
学科主题物理化学
Organic-inorganic hybrid silica monolith based immobilized titanium ion affinity chromatography column for analysis of mitochondrial phosphoproteome
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
2010-09-19
会议地点澳大利亚
页码100/2
部门归属生物技术部
主办者University of New South Wales
英文摘要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
文献类型会议论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/114164
专题中国科学院大连化学物理研究所
通讯作者Zhang LH(张丽华)
推荐引用方式
GB/T 7714
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],2010:100/2.
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