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Ferrous iron is released when purified AcnA is inactivated by superoxide. Purified AcnA (7 μM) was exposed to the xanthine oxidase superoxide-generating system in aerobic buffer. Released Fe2+ was detected by formation of a colored complex with ferrene. Catalase was present to scavenge H2O2; where indicated, SOD was added to scavenge superoxide. Most (>80%) aconitase activity was absent by the first time point.
