title: Proteomic survey of copper-binding proteins in Arabidopsis roots by immobilized metal affinity chromatography and mass spectrometry.
abstract: 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.
authors.affiliation: Institute of BioAgricultural Sciences, Academia Sinica, Taipei, Taiwan, Republic of China.
authors.forename: Cheng-Che S
authors.initials: CC
authors.lastname: Kung
authors.forename: Wei-Ning
authors.initials: WN
authors.lastname: Huang
authors.forename: Yan-Chen
authors.initials: YC
authors.lastname: Huang
authors.forename: Kuo-Chen
authors.initials: KC
authors.lastname: Yeh
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D002846:Chromatography, Affinity; D003300:Copper / Q000378:metabolism*; D013058:Mass Spectrometry; D008667:Metalloproteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D018517:Plant Roots / Q000737:chemistry*; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
chemical_list: D008667:Metalloproteins; D020543:Proteome; D003300:Copper; D015032:Zinc