Pulse radiolysis kinetics of the reaction of hydrated electrons with ferric-, ferrous-, protoporphyrin IX- and apo-myoglobin.

The kinetics of the reaction of hydrated electron (e-aq) with ferric-, ferrous-, metal-free protoporphyrin IX-and apo-myoglobin have been studied by pulse radiolysis so that a direct kinetic measure of the relative reactivities of the heme and the protein part of myoglobin can be made. The second-order association rate constant with ferric Mb is about 3-times that for ApoMb, while ferric Mb, ferrous Mb and protoporphyrin IX-Mb all react at about the same rate, indicating that it is mainly the porphyrin that is the electron-attracting site. The magnitude of the rate constants (8-25 nM-1 X S-1) indicates that the encounter of e-aq with the protein is almost certainly diffusion-controlled. The initial encounter is probably followed by electron migration along parallel paths to the heme and most likely several of the 12 histidine residues. The heme competes very effectively (approx. 70%) with these other sites. The kinetically measured reduction yield of heme is consistent with that found spectrally, indicating that a histidine radical on the protein does not effectively transfer an electron intramolecularly to the heme. The spectral changes found upon the completion of the fast reaction (approx. 40 microseconds) for protoporphyrin IX-Mb and ferrous Mb are consistent with the formation of a porphyrin anion radical. For ApoMb the spectral changes are consistent with the formation of a histidine free radical.