Proteomic survey of copper-binding proteins in Arabidopsis roots by immobilized metal affinity chromatography and mass spectrometry
Proteomic survey of copper-binding proteins in Arabidopsis roots by immobilized metal affinity chromatography and mass spectrometry
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DOI：
10.1002/pmic.200500108
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发表时间：
2006-05-01
期刊：
影响因子：
3.4
通讯作者：
Yeh, Kuo-Chen
中科院分区：
文献类型：
--
作者：
Kung, Cheng-Che S.;Huang, Wei-Ning;Yeh, Kuo-Chen
对于植物来说，铜在低浓度下是至关重要的，但在高浓度下却具有剧毒。植物已经进化出了一套调节铜离子的吸收、分布和利用的机制。这些机制需要铜相互作用蛋白来运输、螯合和隔离铜离子。在这项研究中，我们通过铜固定金属亲和层析（Cu-IMAC）系统地筛选了拟南芥根中的铜相互作用蛋白。我们还比较了拟南芥根金属蛋白质组与 Cu-IMAC 和 Zn-IMAC 的亲和力。从与 Cu-IMAC 具有亲和力的 38 个蛋白点的身份中，鉴定出 35 个独特的蛋白。这些蛋白质的功能分类包括氧化还原/水解反应、氨基酸代谢、谷胱甘肽代谢、磷酸化、翻译机制、膜相关蛋白和营养储存蛋白。预测并评分了潜在的铜相互作用基序。六个候选基序 H-(X)(5)-H、H-(X)(7)-H、H(X)(12)-H、H-(X)(6)-M、M-(X)(7)-H 和 H-(X)(3)-C 存在于 Cu-IMAC 分离蛋白中，频率高于整个拟南芥蛋白质组。
To plants, copper is vitally essential at low concentrations but extremely toxic at elevated concentrations. Plants have evolved a suite of mechanisms that modulate the uptake, distribution, and utilization of copper ions. These mechanisms require copper-interacting proteins for transporting, chelating, and sequestrating copper ions. In this study, we have systematically screened for copper-interacting proteins in Arabidopsis roots via copper-immobilized metal affinity chromatography (Cu-IMAC). We also compared Arabidopsis root metalloproteomes with affinity to Cu-IMAC and Zn-IMAC. From the identities of 38 protein spots with affinity to Cu-IMAC, 35 unique proteins were identified. Functional classification of these proteins includes redox/hydrolytic reactions, amino acid metabolism, glutathione metabolism, phosphorylation, translation machinery, membrane-associated proteins, and vegetative storage proteins. Potential copper-interacting motifs were predicted and scored. Six candidate motifs, H-(X)(5)-H, H-(X)(7)-H, H(X)(12)-H, H-(X)(6)-M, M-(X)(7)-H, and H-(X)(3)-C, are present in Cu-IMAC-isolated proteins with higher frequency than in the whole Arabidopsis proteome.