Flavin-containing polyamine oxidase is a hydrogen peroxide source in the oxidative response to the protein phosphatase inhibitor cantharidin in Zea mays L.
In this study, the specific contribution of polyamine oxidase (PAO), a hydrogen peroxide (H2O2)-producing enzyme, to the oxidative burst induced in maize mesocotyl by the phosphatase inhibitor cantharidin was examined. For this purpose, a pharmacological approach was applied using, either in vitro or in vivo, two strong inhibitors of maize PAO (MPAO), N-prenylagmatine (G3) and its structural analogue Ro5, as well as diphenyleneiodonium (DPI), an inhibitor of the phagocyte NAD(P)H oxidase. DPI was shown to be a good MPAO inhibitor in vitro. G3, Ro5, and DPI were very effective in inhibiting in vivo the extracellular accumulation of H2O2 that is released by mesocotyl segments upon spermidine supply. G3 and Ro5 did not show any inhibition in vitro of either horseradish peroxidase or barley oxalate oxidase. Moreover, G3 and Ro5 did not inhibit the extracellular accumulation of superoxide radical that is released in vivo upon NADH supply. G3, Ro5, and DPI strongly affected H2O2 production induced in maize mesocotyl by cantharidin. Histochemical localization of H2O2 in cantharidin-treated mesocotyl cross-sections revealed an increase of H2O2-specific staining in the epidermal and subepidermal tissues. The effect was also inhibited by G3 and DPI. Moreover, an increase in MPAO activity was observed in the same tissues upon cantharidin treatment. All these data suggest that G3 and Ro5 behave as powerful and selective inhibitors of MPAO activity either in vitro or in vivo and that MPAO activity contributes to a major part of the cantharidin-induced H2O2 synthesis in the apoplastic milieu of maize mesocotyl.
Cona
Alessandra
A
Dipartimento di Biologia, Università degli Studi Roma Tre, Viale Guglielmo Marconi 446, 00146 Rome, Italy.
Rea
Giuseppina
G
Botta
Maurizio
M
Corelli
Federico
F
Federico
Rodolfo
R
Angelini
Riccardo
R
D000376:Agmatine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002193:Cantharidin / Q000494:pharmacology*; D006861:Hydrogen Peroxide / Q000378:metabolism*; D009861:Onium Compounds / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors / Q000187:drug effects*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors; D036226:Seedlings / Q000187:drug effects; D003313:Zea mays / Q000187:drug effects*; D000097667:Polyamine Oxidase
D000376:Agmatine; D002193:Cantharidin; D006861:Hydrogen Peroxide; D009861:Onium Compounds; D000587:Oxidoreductases Acting on CH-NH Group Donors; D010749:Phosphoprotein Phosphatases; D000097667:Polyamine Oxidase; C514183:N-prenylagmatine; C007517:diphenyleneiodonium