Metal ion complexes of apoferritin. Evidence for initial binding in the hydrophilic channels.
Metal binding to the iron storage protein apoferritin is the first step in the process by which iron accumulates within the protein shell. In the present study, the stoichiometry of metal binding to apoferritin in solution has been examined using the probe ions Mn(II), VO(IV), and Cd(II) in conjunction with EPR spectroscopic and cadmium ion selective electrode measurements. Binding studies were carried out with the individual ions, in competition with one another, and in competition with Fe(II), Fe(III), and Tb(III). All three probe ions show binding stoichiometries near 0.3 and 0.7 metal ion per subunit, close to the theoretically predicted values of 0.33 and 0.67 for the binding of one and two metal ions, respectively, per three subunits. These results in conjunction with other data are consistent with the binding of one, and possibly two, metal ions within each of the eight hydrophilic channels which are located on 3-fold axes leading to the interior of the protein. Pairs of cadmium binding sites have been located in these channels by x-ray crystallography (Rice, D. W., Ford, G. C., White, J. L., Smith, J. M. A., and Harrison, P. M. (1983) Adv. Inorg. Biochem. 5, 39-49). The possibility that some metal binding occurs elsewhere on the protein is not precluded by the present data, however. In competition experiments between various metal ions, approximately 0.3 metal ion per subunit is readily displaced implying common binding sites in the channels for all of them. The stoichiometry of Mn(II) displacement by Fe(II) is less clear. Oxidation of Fe(II) to Fe(III) by molecular oxygen in the presence of Mn(II) regenerates some Mn(II) binding on the protein, suggesting migration of iron(III) to other protein sites, or perhaps to core.
D000818:Animals; D001052:Apoferritins / Q000378:metabolism*; D002104:Cadmium / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D005293:Ferritins / Q000031:analogs & derivatives*; D006736:Horses; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000378:metabolism; D007700:Kinetics; D046911:Macromolecular Substances; D008345:Manganese / Q000378:metabolism; D008670:Metals / Q000378:metabolism*; D011485:Protein Binding; D013154:Spleen / Q000378:metabolism; D013725:Terbium / Q000378:metabolism
D046911:Macromolecular Substances; D008670:Metals; D002104:Cadmium; D013725:Terbium; D008345:Manganese; D005293:Ferritins; D001052:Apoferritins; D007501:Iron