Luminol peroxidation catalyzed by human isoferritins.
The role of ferritin in catalyzing the oxidation of luminol with the production of chemiluminescence was investigated. The effect of pH was compared to its effect on K3Fe(CN)6-catalyzed oxidation and different pH optima were recorded for the two catalysts. The ferrous iron chelator, bipyridyl, enhanced the production of chemiluminescence catalyzed by FeSO4 and ferritin but had little effect on the K3Fe(CN)6-catalyzed reaction. Desferal reduced the level of chemiluminescence in the presence of FeSO4 and ferritin but was a much more effective inhibitor of chemiluminescence catalyzed by K3Fe(CN)6. The hydroxyl radical scavenger, mannitol, had little effect upon light production whereas superoxide dismutase inhibited light production. The addition of antihuman spleen ferritin completely inhibited activity. The catalytic activity of both H and L rich ferritins was affected by iron content. Activity increased until the Fe/protein ratio reached 0.04 micrograms Fe/micrograms protein and then decreased with increasing iron content. Thus activity is controlled by the iron content of the molecule and influenced by its subunit composition as is the uptake of iron into ferritin. These findings suggest that ferroxidation by ferritin is associated with the ability to generate radicals of the nitrogenous base luminol with the production of chemiluminescence. Although activity is greatest at alkaline pH there is significant activity at pH 7.4. Ferritin therefore may be able to generate free radical reactions in vivo with the acidic isoferritin being most active.
Henley
R
R
Department of Medical Biochemistry, University Hospital of Wales, Health Park, Cardiff, United Kingdom.
Worwood
M
M
D015082:2,2'-Dipyridyl / Q000494:pharmacology; D003676:Deferoxamine / Q000494:pharmacology; D005292:Ferricyanides; D005293:Ferritins / Q000378:metabolism*; D016166:Free Radical Scavengers; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008163:Luminescent Measurements; D008165:Luminol / Q000378:metabolism*; D010084:Oxidation-Reduction; D013154:Spleen / Q000378:metabolism; D013482:Superoxide Dismutase / Q000494:pharmacology
D005292:Ferricyanides; D016166:Free Radical Scavengers; D015082:2,2'-Dipyridyl; D008165:Luminol; D005293:Ferritins; D013482:Superoxide Dismutase; D003676:Deferoxamine; C028033:potassium ferricyanide