title: Exposure of aconitase to smoking-related oxidants results in iron loss and increased iron response protein-1 activity: potential mechanisms for iron accumulation in human arterial cells.
abstract: Smokers have an elevated risk of cardiovascular disease, but the origin(s) of this increased risk are incompletely defined. Evidence supports an accumulation of the oxidant-generating enzyme myeloperoxidase (MPO) in the inflamed artery wall, and smokers have high levels of SCN(-), a preferred MPO substrate, with this resulting in HOSCN formation. We hypothesised that HOSCN, a thiol-specific oxidant may target the iron-sulphur cluster of aconitase (both isolated, and within primary human coronary artery endothelial cells; HCAEC) resulting in enzyme dysfunction, release of iron, and conversion of the cytosolic isoform to iron response protein-1, which regulates intracellular iron levels. We show that exposure of isolated aconitase to increasing concentrations of HOSCN releases iron from the aconitase [Fe-S]4 cluster, and decreases enzyme activity. This is associated with protein thiol loss and modification of specific Cys residues in, and around, the [Fe-S]4 cluster. Exposure of HCAEC to HOSCN resulted in increased intracellular levels of chelatable iron, loss of aconitase activity and increased iron response protein-1 (IRP-1) activity. These data indicate HOSCN, an oxidant associated with oxidative stress in smokers, can induce aconitase dysfunction in human endothelial cells via Cys oxidation, damage to the [Fe-S]4 cluster, iron release and generation of IRP-1 activity, which modulates ferritin protein levels and results in dysregulation of iron metabolism. These data may rationalise, in part, the presence of increased levels of iron in human atherosclerotic lesions and contribute to increased oxidative damage and endothelial cell dysfunction in smokers. Similar reactions may occur at other sites of inflammation.
authors.affiliation: The Heart Research Institute, 7 Eliza Street, Newtown, Sydney, NSW, 2042, Australia.|Faculty of Medicine, University of Sydney, Sydney, NSW, 2006, Australia.
authors.forename: Jihan
authors.initials: J
authors.lastname: Talib
authors.affiliation: The Heart Research Institute, 7 Eliza Street, Newtown, Sydney, NSW, 2042, Australia. davies@sund.ku.dk.|Faculty of Medicine, University of Sydney, Sydney, NSW, 2006, Australia. davies@sund.ku.dk.|Department of Biomedical Sciences, Panum Institute, University of Copenhagen, 2200, Copenhagen, Denmark. davies@sund.ku.dk.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Davies
mesh_terms: D000154:Aconitate Hydratase / Q000737:chemistry / Q000378:metabolism*; D002478:Cells, Cultured; D003331:Coronary Vessels / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology; D007501:Iron / Q000378:metabolism; D000090463:Iron Deficiencies; D035925:Iron-Regulatory Proteins / Q000378:metabolism*; D016877:Oxidants / Q000737:chemistry / Q000378:metabolism*; D012907:Smoking / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013861:Thiocyanates / Q000494:pharmacology
chemical_list: D035925:Iron-Regulatory Proteins; D016877:Oxidants; D013861:Thiocyanates; C517667:hypothiocyanous acid; D006997:Hypochlorous Acid; D007501:Iron; D000154:Aconitate Hydratase
keywords: Aconitase; Cardiovascular disease; Hypothiocyanous acid; Inflammation; Iron release; Iron response protein-1; Myeloperoxidase; Thiol oxidation
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