Effects of Ionizable Groups on the Redox Potentials of Cytochrome c3 from D. vulgaris Miyazaki F

摘要

The p2H dependence of the NMR chemical shifts of the proton signals of heme methyl groups and ionizable groups in the vicinity of the heme were investigated. The p2H titration of heme methyl signals in four macroscopic oxidation states by saturation tranfer method was performed in the range between p2H 5.2 and 9.0. While the p2H dependence of the heme methyl resonance in fully oxidized state was small, most resonances in the intermediate oxidation states showed certain shifts. Particularly, methyl resonances of heme 1 (sequential heme numbering) exhibited sharp p2H dependence in acidic range. モ-CH2 of the propionate of hemes 1 and 4 were titrated in the range of p2H 4.5-9.0. Only the 6-propionate group of heme 1 was protonated in this p2H range and its titration curve was similar to those of methyl resonances of heme 1 in intermediate oxidation states. Analysis of the microscopic redox potentials showed that they change depending on p2H. The ionizable groups responsible for the p2H dependence of these potentials are 6-propionate of heme 1 in acidic range and His 67 in basic range.