title: Divalent metal ions in plant mitochondria and their role in interactions with proteins and oxidative stress-induced damage to respiratory function.
abstract: Understanding the metal ion content of plant mitochondria and metal ion interactions with the proteome are vital for insights into both normal respiratory function and the process of protein damage during oxidative stress. We have analyzed the metal content of isolated Arabidopsis (Arabidopsis thaliana) mitochondria, revealing a 26:8:6:1 molar ratio for iron:zinc:copper:manganese and trace amounts of cobalt and molybdenum. We show that selective changes occur in mitochondrial copper and iron content following in vivo and in vitro oxidative stresses. Immobilized metal affinity chromatography charged with Cu(2+), Zn(2+), and Co(2+) was used to identify over 100 mitochondrial proteins with metal-binding properties. There were strong correlations between the sets of immobilized metal affinity chromatography-interacting proteins, proteins predicted to contain metal-binding motifs, and protein sets known to be oxidized or degraded during abiotic stress. Mitochondrial respiratory chain pathways and matrix enzymes varied widely in their susceptibility to metal-induced loss of function, showing the selectivity of the process. A detailed study of oxidized residues and predicted metal interaction sites in the tricarboxylic acid cycle enzyme aconitase identified selective oxidation of residues in the active site and showed an approach for broader screening of functionally significant oxidation events in the mitochondrial proteome.
authors.affiliation: Australian Research Council Centre of Excellence in Plant Energy Biology, M316, University of Western Australia, Crawley, Western Australia 6009, Australia.
authors.forename: Yew-Foon
authors.initials: YF
authors.lastname: Tan
authors.forename: Nicholas
authors.initials: N
authors.lastname: O'Toole
authors.forename: Nicolas L
authors.initials: NL
authors.lastname: Taylor
authors.forename: A Harvey
authors.initials: AH
authors.lastname: Millar
mesh_terms: D017360:Arabidopsis / Q000737:chemistry*; D002413:Cations, Divalent / Q000378:metabolism; D002846:Chromatography, Affinity; D006706:Homeostasis; D019216:Metals, Heavy / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010940:Plant Proteins / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D019216:Metals, Heavy; D024101:Mitochondrial Proteins; D010940:Plant Proteins