CYTOCHROME BO FROM ESCHERICHIA-COLI DOES NOT EXHIBIT THE SAME PROTON-TRANSFER CHARACTERISTICS AS THE BOVINE CYTOCHROME-C-OXIDASE DURING OXYGEN REDUCTION

The reaction where fully reduced cytochrome bo from E coli partially reduces dioxygen has been characterized with respect to the kinetics of the associated proton uptake, and with respect to the pH- and D2O-sensitivity of the electron transfer reactions. A monophasic proton uptake with a rate constant of about 8 x 10(3) s-1 and a stoichiometry of 0.8 H+/bo were recorded, using the indicator dye, Cresol red, at pH 8.2. The electron transfer reactions were independent of pH in the range 6.0-9.5 and were not affected by exchanging H2O to D2O as solvent. Comparison of these results with those obtained in an earlier investigation of the bovine cytochrome c oxidase [(1992) Biochemistry 31, 11853-11859], indicates differences between the two oxidases with respect to the role of protons in oxygen reduction and/or the mechanism of proton uptake from the medium.