title: Copper depletion increases the mitochondrial-associated SOD1 in neuronal cells.
abstract: The role of copper in the toxicity of mutant copper-dependent enzyme superoxide dismutase (SOD1) found in patients affected with the familial form of amyotrophic lateral sclerosis (fALS) is widely debated. Here we report that treatment of human neuroblastoma cells SH-SY5Y with a specific copper chelator, triethylene tetramine (Trien) induces the decrease of intracellular copper level, paralleled by decreased activity of SOD1. A comparable effect is observed in mouse NSC-34-derived cells, a motoneuronal model, transfected for the inducible expression of either wild-type or G93A mutant human SOD1, one of the mutations associated with fALS. In both cell types, the drop of SOD1 activity is not paralleled by the same extent of decrease in SOD1 protein content. This discrepancy can be explained by the occurrence of a fraction of copper-free SOD1 upon copper depletion, which is demonstrated by the partial recovery of the enzyme activity after the addition of copper sulphate to homogenates of SH-SY5Y cells. Furthermore, copper depletion produces the enrichment of the physiological mitochondrial fraction of SOD1 protein, in both cells models. However, increasing the fraction of mitochondrial, possibly copper-free, mutant human SOD1 does not further alter mitochondrial morphology in NSC-34-derived cells. Thus, copper deficiency is not a factor which may worsen mitochondrial damage, which is one of the earliest events in fALS associated with mutant SOD1.
authors.affiliation: Department of Biology, Tor Vergata University of Rome, Via della Ricerca Scientifica 1, 00133, Rome, Italy.
authors.forename: Mario
authors.initials: M
authors.lastname: Arciello
authors.forename: Concetta Rosa
authors.initials: CR
authors.lastname: Capo
authors.forename: Sara
authors.initials: S
authors.lastname: D'Annibale
authors.forename: Mauro
authors.initials: M
authors.lastname: Cozzolino
authors.forename: Alberto
authors.initials: A
authors.lastname: Ferri
authors.forename: Maria Teresa
authors.initials: MT
authors.lastname: Carrì
authors.forename: Luisa
authors.initials: L
authors.lastname: Rossi
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D045744:Cell Line, Tumor; D003300:Copper / Q000172:deficiency*; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism*; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism*; D000072105:Superoxide Dismutase-1; D014266:Trientine / Q000494:pharmacology
chemical_list: C000606290:SOD1 protein, human; D003300:Copper; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1; D014266:Trientine