title: A role for rubredoxin in oxidative stress protection in Desulfovibrio vulgaris: catalytic electron transfer to rubrerythrin and two-iron superoxide reductase.
abstract: Desulfovibrio vulgaris rubredoxin, which contains a single [Fe(SCys)4] site, is shown to be a catalytically competent electron donor to two enzymes from the same organism, namely, rubrerythrin and two-iron superoxide reductase (a.k.a. rubredoxin oxidoreductase or desulfoferrodoxin). These two enzymes have been implicated in catalytic reduction of hydrogen peroxide and superoxide, respectively, during periods of oxidative stress in D. vulgaris, but their proximal electron donors had not been characterized. We further demonstrate the incorrectness of a previous report that rubredoxin is not an electron donor to the superoxide reductase and describe convenient assays for demonstrating the catalytic competence of all three proteins in their respective functions. Rubrerythrin is shown to be an efficient rubredoxin peroxidase in which the rubedoxin:hydrogen peroxide redox stoichiometry is 2:1 mol:mol. Using spinach ferredoxin-NADP+ oxidoreductase (FNR) as an artificial, but proficient, NADPH:rubredoxin reductase, rubredoxin was further found to catalyze rapid and complete reduction of all Fe3+ to Fe2+ in rubrerythrin by NADPH under anaerobic conditions. The combined system, FNR/rubredoxin/rubrerythrin, was shown to function as a catalytically competent NADPH peroxidase. Another small rubredoxin-like D. vulgaris protein, Rdl, could not substitute for rubredoxin as a peroxidase substrate of rubrerythrin. Similarly, D. vulgaris rubredoxin was demonstrated to efficiently catalyze reduction of D. vulgaris two-iron superoxide reductase and, when combined with FNR, to function as an NADPH:superoxide oxidoreductase. We suggest that, during periods of oxidative stress, rubredoxin could divert electron flow from the electron transport chain of D. vulgaris to rubrerythrin and superoxide reductase, thereby simultaneously protecting autoxidizable redox enzymes and lowering intracellular hydrogen peroxide and superoxide levels.
authors.affiliation: Department of Chemistry, University of Georgia, Athens, Georgia 30602, USA.
authors.forename: E D
authors.initials: ED
authors.lastname: Coulter
authors.forename: D M
authors.initials: DM
authors.lastname: Kurtz
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D001665:Binding Sites; D002384:Catalysis; D016969:Desulfovibrio vulgaris / Q000201:enzymology / Q000378:metabolism*; D019281:Dimerization; D004579:Electron Transport; D004583:Electrons; D005288:Ferredoxins / Q000378:metabolism*; D006422:Hemerythrin; D006861:Hydrogen Peroxide / Q000378:metabolism; D007700:Kinetics; D009247:NADH, NADPH Oxidoreductases / Q000378:metabolism*; D009249:NADP / Q000378:metabolism; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010544:Peroxidases / Q000378:metabolism; D019163:Reducing Agents / Q000378:metabolism; D012416:Rubredoxins / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D005288:Ferredoxins; D006422:Hemerythrin; D019163:Reducing Agents; D012416:Rubredoxins; C055857:rubrerythrins; D009249:NADP; D006861:Hydrogen Peroxide; D010544:Peroxidases; C045195:NADPH peroxidase; C023091:rubredoxin-NAD+ reductase; D009247:NADH, NADPH Oxidoreductases