Photoinduced oxidation of microperoxidase-8: Generation of ferryl and cation-radical porphyrins

The electron-transfer reactivity of microperoxidase-8 (MP8), the heme octapeptide derived from enzymatic cleavage of cytochrome c, has been studied by nanosecond flash photolysis methods. Ferric MP8 is rapidly oxidized by photogenerated Ru(bpy)(3)(3+) in acidic solutions to a ferric cation-radical porphyrin (k similar to 5.6 x 10(9) M(-1) s(-1)); the oxidation product in alkaline solutions is ferryl MP8 (k similar to 2.2 x 10(9) M(-1) s(-1)). Numerical simulations of the kinetics for the direct oxidation of ferric to ferryl MP8 predict a marked pH effect on the rate of reaction in alkaline solutions; however, only a very weak pH dependence is observed in the range 7-8.5, indicating that the ferryl species is generated by intramolecular electron transfer within a ferric cation-radical porphyrin. Transient spectra taken between pH 6 and 8.5 show increasing ferryl absorption as the pH is increased, demonstrating a pH-dependent equilibrium between the two oxidized forms of MP8.