Reaction of the desulfoferrodoxin from Desulfoarculus baarsii with superoxide anion. Evidence for a superoxide reductase activity.
Desulfoferrodoxin is a small protein found in sulfate-reducing bacteria that contains two independent mononuclear iron centers, one ferric and one ferrous. Expression of desulfoferrodoxin from Desulfoarculus baarsii has been reported to functionally complement a superoxide dismutase deficient Escherichia coli strain. To elucidate by which mechanism desulfoferrodoxin could substitute for superoxide dismutase in E. coli, we have purified the recombinant protein and studied its reactivity toward O-(2). Desulfoferrodoxin exhibited only a weak superoxide dismutase activity (20 units mg(-1)) that could hardly account for its antioxidant properties. UV-visible and electron paramagnetic resonance spectroscopy studies revealed that the ferrous center of desulfoferrodoxin could specifically and efficiently reduce O-(2), with a rate constant of 6-7 x 10(8) M(-1) s(-1). In addition, we showed that membrane and cytoplasmic E. coli protein extracts, using NADH and NADPH as electron donors, could reduce the O-(2) oxidized form of desulfoferrodoxin. Taken together, these results strongly suggest that desulfoferrodoxin behaves as a superoxide reductase enzyme and thus provide new insights into the biological mechanisms designed for protection from oxidative stresses.
Lombard
M
M
Laboratoire de Chimie et Biochimie des Centres Redox Biologiques, DBMS-CEA/CNRS/Université Joseph Fourier, 17 Avenue des Martyrs, 38054 Grenoble, Cedex 9, France.
Fontecave
M
M
Touati
D
D
Nivière
V
V
D000595:Amino Acid Sequence; D003574:Cytochrome c Group / Q000378:metabolism; D003901:Desulfovibrio / Q000201:enzymology*; D004926:Escherichia coli / Q000235:genetics; D005288:Ferredoxins / Q000235:genetics / Q000378:metabolism*; D005296:Ferrous Compounds; D007501:Iron / Q000032:analysis; D008956:Models, Chemical; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011994:Recombinant Proteins / Q000378:metabolism; D013053:Spectrophotometry; D013482:Superoxide Dismutase / Q000378:metabolism; D013481:Superoxides / Q000378:metabolism*
D003574:Cytochrome c Group; D005288:Ferredoxins; D005296:Ferrous Compounds; D011994:Recombinant Proteins; C066602:desulfoferrodoxin; D013481:Superoxides; D007501:Iron; D013482:Superoxide Dismutase