Hydrogen peroxide damages the zinc-binding site of zinc-deficient Cu,Zn superoxide dismutase.
Mutations in Cu,Zn superoxide dismutase (Cu,Zn SOD) account for approximately 20% of cases of familial amyotrophic lateral sclerosis (ALS), a late-onset neurodegenerative disease affecting motor neurons. These mutations decrease protein stability and lower zinc affinity. Zinc-deficient SOD (Cu,E SOD) has altered redox activities and is toxic to motor neurons in vitro. Using bovine SOD, we studied the effects of hydrogen peroxide (H(2)O(2)) on Cu,E SOD and Cu,Zn SOD. Hydrogen peroxide treatment of Cu,E SOD inactivated zinc binding activity six times faster than superoxide dismutase activity, whereas inactivation of dismutase activity occurred at the same rate for both Cu,Zn SOD and Cu,E SOD. Zinc binding by Cu,E SOD was also damaged by simultaneous generation of superoxide and hydrogen peroxide by xanthine oxidase plus xanthine. Although urate, xanthine, and ascorbate can protect superoxide dismutase activity of Cu,Zn SOD from inactivation, they were not effective at protecting Cu,E SOD. Hydrogen peroxide induced subtle changes in the tertiary structure but not the secondary structure of Cu,E SOD as detected by near and far UV circular dichroism. Our results suggest that low levels of hydrogen peroxide could potentially enhance the toxicity of zinc deficient SOD to motor neurons in ALS by rendering zinc loss from SOD irreversible.
Sampson
J B
JB
Department of Anesthesiology, University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
Beckman
J S
JS
D000690:Amyotrophic Lateral Sclerosis / Q000201:enzymology / Q000209:etiology / Q000235:genetics; D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002942:Circular Dichroism; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D066298:In Vitro Techniques; D007700:Kinetics; D009046:Motor Neurons / Q000201:enzymology; D009154:Mutation; D010084:Oxidation-Reduction; D017433:Protein Structure, Secondary; D013482:Superoxide Dismutase / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics*; D015032:Zinc / Q000378:metabolism*
D004791:Enzyme Inhibitors; D006861:Hydrogen Peroxide; D013482:Superoxide Dismutase; D015032:Zinc