Oxidation and reduction of cytochrome c bound to the phosphoprotein phosvitin.

The oxidation-reduction reactions and structural characteristics of phosvitin-bound cytochrome c were examined at various ratios of cytochrome c to phosvitin. At binding ratios below half the maximum, the rate constants for the oxidation reactions with cytochrome c oxidase and ferricyanide and the rate constants for the reduction reactions with cytochrome b2 and ascorbate were low, but at higher ratios these rate constants gradually increased to that of free cytochrome c and, in particular, the rate constant for oxidation by cytochrome c oxidase was raised to two to three times that of the free form. This binding-ratio dependence of the rate constants for the oxidation and reduction reactions was different from that of the net charge of the cytochrome c-phosvitin complex, implying that the negative charges of phosvitin are unlikely to modulate the rates. In contrast, the broadening of the NMR signals for the heme and methionine-80 methyl groups and the conformational transition in the vicinity of the heme moiety on change from the native to the cyanide-bound or urea-denatured form of cytochrome c showed a similar binding-ratio dependence to the rate constants for the oxidation and reduction reactions. Since the conformation and electronic structure in the heme environment of ferric and ferrous cytochromes c were not changed significantly by binding to phosvitin, and since the binding strength of cytochrome c to phosvitin at binding ratios below half the maximum is different from that at higher ratios, these findings suggest that a difference in the movement of cytochrome c in its complex with phosvitin may modulate its oxidation-reduction reactions.