Dynamic equilibria in iron uptake and release by ferritin.
The function of ferritins is to store and release ferrous iron. During oxidative iron uptake, ferritin tends to lower Fe2+ concentration, thus competing with Fenton reactions and limiting hydroxy radical generation. When ferritin functions as a releasing iron agent, the oxidative damage is stimulated. The antioxidant versus pro-oxidant functions of ferritin are studied here in the presence of Fe2+, oxygen and reducing agents. The Fe(2+)-dependent radical damage is measured using supercoiled DNA as a target molecule. The relaxation of supercoiled DNA is quantitatively correlated to the concentration of exogenous Fe2+, providing an indirect assay for free Fe2+. After addition of ferrous iron to ferritin, Fe2+ is actively taken up and asymptotically reaches a stable concentration of 1-5 microM. Comparable equilibrium concentrations are found with plant or horse spleen ferritins, or their apoferritins. After addition of ascorbate, iron release is observed using ferrozine as an iron scavenger. Rates of iron release are dependent on ascorbate concentration. They are about 10 times larger with pea ferritin than with horse ferritin. In the absence of ferrozine, the reaction of ascorbate with ferritins produces a wave of radical damage; its amplitude increases with increased ascorbate concentrations with plant ferritin; the damage is weaker with horse ferritin and less dependent on ascorbate concentrations.
Laulhère
J P
JP
Chimie Bioinorganioque, LEDSS, Université J. Fourier, URA CNRS 0332, Grenoble, France.
Barcelò
F
F
Fontecave
M
M
D001205:Ascorbic Acid / Q000494:pharmacology; D004278:DNA, Superhelical; D005293:Ferritins / Q000378:metabolism*; D005296:Ferrous Compounds / Q000378:metabolism*; D005609:Free Radicals; D007700:Kinetics
D004278:DNA, Superhelical; D005296:Ferrous Compounds; D005609:Free Radicals; D005293:Ferritins; D001205:Ascorbic Acid