Cytochrome b−245 of the Neutrophil Superoxide-generating System Contains Two Nonidentical Hemes

Analysis of potentiometric titrations of the cytochrome b-245 from a X+ chronic granulomatous disease patient with an Arg54 --> Ser mutation in gp91phox indicates that the mutant form of the cytochrome contains two nonidentical hemes with midpoint potentials of Em7 = -220 and Em7 = -300 mV. In the light of this information, reanalysis of redox titrations of wild-type cytochrome b-245 implies that it probably also contains two separate heme centers with midpoint potentials of Em7 = -225 and Em7 = -265 mV. The effect of the Arg54 --> Ser substitution is to reduce the midpoint potential of one of the heme centers by approximately 35 mV and suggests possible interaction between Arg54 and a heme propionate side chain.