Stimulation of the ferroxidase activity of ceruloplasmin during iron loading into ferritin.
Ceruloplasmin purified from horse serum was rapidly reduced upon addition of increasing equivalents of ferrous iron, generating an electronically and conformationally distinct form. This form of ceruloplasmin was characterized by significant (80%) loss of EPR detectable type I and type II copper(II), complete loss of visible absorbance at 610 nm, as well as decreased hydrophobic surface area. The reduced form of ceruloplasmin slowly reduced molecular oxygen to complete its catalytic cycle. The presence of varied concentrations of apoferritin, but not apotransferrin, significantly enhanced the rate of ceruloplasmin oxidation. The magnitude of this stimulatory effect increased as the molar ratio of ceruloplasmin to apoferritin approached 1.0, shown previously to be the optimum ratio for loading iron into ferritin. The rate of ferrous iron oxidation by ceruloplasmin was significantly stimulated by the presence of apoferritin; however, apotransferrin had no effect. The length of time required for ceruloplasmin to oxidize all the iron and return to the native form of the enzyme was also affected by the concentration of iron. In addition, the rate of iron loading into ferritin was dependent upon ferrous iron concentration. These results provide evidence for the formation of a specific complex between the reduced form of ceruloplasmin and apoferritin and that reduction of ceruloplasmin by ferrous iron may be the signal for complex formation.
Reilly
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Biotechnology Center, Utah State University, Logan, Utah 84322-4705, USA.
Aust
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D000818:Animals; D001052:Apoferritins / Q000378:metabolism; D002570:Ceruloplasmin / Q000378:metabolism*; D003300:Copper / Q000378:metabolism; D004578:Electron Spin Resonance Spectroscopy; D005293:Ferritins / Q000378:metabolism*; D006736:Horses; D007501:Iron / Q000378:metabolism*; D010084:Oxidation-Reduction; D013050:Spectrometry, Fluorescence; D013053:Spectrophotometry
D003300:Copper; D005293:Ferritins; D001052:Apoferritins; D007501:Iron; D002570:Ceruloplasmin