A pulse-radiolysis study of cytochrome c3. Kinetics of the reduction of cytochrome c3 by methyl viologen radicals and the characterisation of the redox properties of cytochrome c3 from Desulfovibrio vulgaris (Hildenborough).
1. Pulse-radiolysis experiments were performed in the presence of methyl viologen and cytochrome c3. After the pulse, methyl viologen radicals are formed and the kinetics of these radicals with cytochrome c3 are studied, The reaction between cytochrome c3 and methyl viologen radicals (MV+) is diffusion controlled. The ionic strength dependence and the pH-dependence of this reaction were studied. From the ionic strength dependence (at pH 7.8) we found that the net charge of the fully oxidized cytochrome c3 molecule was Z = + 4.7 +/- 0.7. 2. After the pulse an equilibrium is reached for the reaction of MV+ with cytochrome c3. From this equilibrium an apparent midpoint potential can be obtained. The apparent midpoint potential of this multihaem molecule was found to depend on the degree of reduction, alpha. With the help of the Nernst equation an empirical equation is obtained to describe this dependence of the midpoint potential: E0 = - 0.250 - 0.088 alpha (in V). 3. An estimation is made of the energy of interaction between the haems due to electrostatic interactions (delta epsilon less than 32 mV) and due to ionic strength effects (- 12 mV less than delta epsilon less than 26 mV). The results suggest that the redox properties of the individual haems in the cytochrome c3 molecule are dependent on the degree of reduction of the other haems in the molecule. 4. The reaction of cytochrome c3 with MV+ or with ethanol radicals (EtOH) has been compared with the reactions of horse-heart cytochrome c and of metmyoglobin with the same radicals. The reaction of MV+ or EtOH with horse-heart cytochrome c is found to be diffusion controlled; the reactions with metmyoglobin on the other hand are most probably controlled by an activation energy.
D000818:Animals; D001426:Bacterial Proteins / Q000302:isolation & purification; D055598:Chemical Phenomena; D002621:Chemistry; D003574:Cytochrome c Group / Q000302:isolation & purification* / Q000378:metabolism; D003901:Desulfovibrio / Q000201:enzymology*; D005609:Free Radicals; D006736:Horses; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008786:Metmyoglobin / Q000378:metabolism; D010084:Oxidation-Reduction / Q000187:drug effects; D010269:Paraquat / Q000494:pharmacology*
D001426:Bacterial Proteins; D003574:Cytochrome c Group; D005609:Free Radicals; D008786:Metmyoglobin; C017165:cytochrome c(3); D010269:Paraquat