Wild-type superoxide dismutase acquires binding and toxic properties of ALS-linked mutant forms through oxidation.
Recent studies suggest that superoxide dismutase (SOD1) may represent a major target of oxidative damage in neurodegenerative diseases. To test the possibility that oxidized species of wild-type (WT) SOD1 might be involved in pathogenic processes, we analyzed the properties of the WT human SOD1 protein after its oxidation in vivo or in vitro by hydrogen peroxide (H2O2) treatment. Using transfected Neuro2a cells expressing WT or amyotrophic lateral sclerosis-linked SOD1 species, we show that exposure to H2O2 modifies the properties of WT SOD1. Western blot analysis of immunoprecipitates from cell lysates revealed that, like mutant SOD1, oxidized WT SOD1 can be conjugated with poly-ubiquitin and can interact with Hsp70. Chromogranin B, a neurosecretory protein that interacts with mutant SOD1 but not with WT SOD1, was co-immunoprecipitated with oxidized WT SOD1 from lysates of Neuro2a cells treated with H2O2. Treatment of microglial cells (line BV2) with either oxidized WT SOD1 or mutant SOD1 recombinant proteins induced tumor necrosis factor-alpha and inducible nitric oxide synthase. Furthermore, exposure of cultured motor neurons to oxidized WT SOD1 caused dose-dependent cell death like mutant SOD1 proteins. These results suggest that WT SOD1 may acquire binding and toxic properties of mutant forms of SOD1 through oxidative damage.
Ezzi
Samer Abou
SA
Department of Anatomy and Physiology, Laval University, Research Centre of CHUL, Québec, Canada.
Urushitani
Makoto
M
Julien
Jean-Pierre
JP
D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics*; D000818:Animals; D016923:Cell Death / Q000235:genetics / Q000502:physiology; D002478:Cells, Cultured; D053378:Chromogranin B / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D015151:Immunoblotting; D047468:Immunoprecipitation; D051379:Mice; D009046:Motor Neurons / Q000502:physiology; D009154:Mutation / Q000502:physiology; D010084:Oxidation-Reduction; D010957:Plasmids / Q000235:genetics; D011485:Protein Binding / Q000235:genetics; D017510:Protein Folding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013116:Spinal Cord / Q000166:cytology; D013347:Subcellular Fractions / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics* / Q000378:metabolism / Q000633:toxicity*; D000072105:Superoxide Dismutase-1; D014162:Transfection; D014451:Ubiquinone / Q000378:metabolism
D053378:Chromogranin B; D011994:Recombinant Proteins; C000606290:SOD1 protein, human; D014451:Ubiquinone; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1