HYDROXYL RADICAL PRODUCTION IN THE REACTIONS OF COPPER-CONTAINING AMINE OXIDASES WITH SUBSTRATES

Solutions of porcine kidney diamine oxidase, PKDAO, and bovine plasma amine oxidase, BPAO, were saturated with the spin-trapping agent alpha-phenyl-N-t-butylnitrone, PBN, and incubated with cadaverine or benzylamine substrate, respectively, under aerobic conditions. EPR spectra due to trapped hydroxyl radicals were obtained for both enzymes with no evidence of superoxide formation. Under anaerobic conditions, hydroxyl radicals were formed only when H2O2 was present as well as substrate. Catalase prevented hydroxyl radical formation by PKDAO but not BPAO. The results indicate that hydroxyl radical is produced in the reaction of the product H2O2 with the reduced enzymes and therefore may be important in turnover-related enzyme degradation, but is not a true reaction intermediate.