Prooxidant action of desferrioxamine: enhancement of alkaline phosphatase inactivation by interaction with ascorbate system.
Desferrioxamine (DFO) nearly doubles alkaline phosphatase oxidative inactivation by the ascorbate system. The effect is dependent on ascorbate and desferrioxamine concentrations, exhibiting in both cases a saturation mechanism. Conversion of desferrioxamine to ferrioxamine abolishes the prooxidant action. Desferrioxamine also increases ascorbate-dependent oxygen consumption and nitroblue tetrazolium reduction. Superoxide dismutase, which blocks the desferrioxamine enhancing effect on enzyme inactivation, markedly slows down nitroblue tetrazolium reduction as well as oxygen consumption by ascorbate plus desferrioxamine, while it fails to protect against the ascorbate system alone. Therefore, in the presence of desferrioxamine, the metal-catalyzed ascorbate autooxidation becomes superoxide-dependent and thus inhibitable by superoxide dismutase. Catalase, peroxidase, and ascorbate oxidase protect alkaline phosphatase from inactivation by both ascorbate and ascorbate-desferrioxamine systems. Hemin shields the enzyme from ascorbate plus DFO attack but not from ascorbate alone. In air-saturated solution, desferrioxamine seems to mediate one electron transfer from ascorbate to oxygen, generating superoxide anions, which can either trigger a Fenton reaction or produce desferal nitroxide radicals. In the absence of oxygen, ascorbate alone is ineffective, but the ascorbate plus desferrioxamine system still inactivates the enzyme; catalase, peroxidase, and ascorbate oxidase, but not superoxide dismutase, afford protection.
Mordente
A
A
Istituto di Chimica Biologica, Università Cattolica del S. Cuore, Facoltà di Medicina, Rome, Italy.
Meucci
E
E
Miggiano
G A
GA
Martorana
G E
GE
D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors*; D001205:Ascorbic Acid / Q000494:pharmacology*; D003676:Deferoxamine / Q000494:pharmacology*; D005609:Free Radicals; D006878:Hydroxides; D017665:Hydroxyl Radical; D007700:Kinetics; D009580:Nitroblue Tetrazolium / Q000378:metabolism; D010084:Oxidation-Reduction; D013482:Superoxide Dismutase / Q000494:pharmacology
D005609:Free Radicals; D006878:Hydroxides; D009580:Nitroblue Tetrazolium; D017665:Hydroxyl Radical; D013482:Superoxide Dismutase; D000469:Alkaline Phosphatase; D003676:Deferoxamine; D001205:Ascorbic Acid