Electrostatic interaction of cytochrome c with cytochrome c1 and cytochrome oxidase.
The reactions of horse heart cytochrome c with succinate-cytochrome c reductase and cytochrome oxidase were studied as a function of ionic strength using both spectrophotometric and oxygen electrode assay techniques. The kinetic parameter Vmax/Km for both reactions decreased very rapidly as the ionic strength was increased, indicating that electrostatic interactions were important to the reactions. A new semiempirical relationship for the electrostatic energy of interaction between cytochrome c and its oxidation-reduction partners was developed, in which specific complementary charge-pair interactions between lysine amino groups on cytochrome c and negatively charged carboxylate groups on the other protein are assumed to dominate the interaction. The contribution of individual cytochrome c lysine amino groups to the electrostatic interaction was estimated from the decrease in reaction rate caused by specific modification of the lysine amino groups by reagents that change the charge to 0 or -1. These estimates range from -0.9 kcal/mol for lysines immediately surrounding the heme crevice of cytochrome c to 0 kcal/mol for lysines well removed from the heme crevice region. The semiempirical relationship for the total electrostatic energy of interaction was in quantitative agreement with the experimental ionic strength dependence of the reaction rates when the parameters were based on the specific lysine modification results. The electrostatic energies of interaction between cytochrome c and its reductase and oxidase were nearly the same, providing additional evidence that the two reactions take place at similar sites on cytochrome c.
D000818:Animals; D002151:Calorimetry; D003574:Cytochrome c Group / Q000031:analogs & derivatives* / Q000378:metabolism*; D003575:Cytochromes c1 / Q000378:metabolism*; D003576:Electron Transport Complex IV / Q000378:metabolism*; D006418:Heme / Q000378:metabolism; D006736:Horses; D007700:Kinetics; D008239:Lysine; D009206:Myocardium / Q000378:metabolism; D009994:Osmolar Concentration; D011048:Polarography; D013367:Submitochondrial Particles / Q000378:metabolism; D013384:Succinate Cytochrome c Oxidoreductase / Q000378:metabolism
D003574:Cytochrome c Group; D006418:Heme; D003575:Cytochromes c1; D013384:Succinate Cytochrome c Oxidoreductase; D003576:Electron Transport Complex IV; D008239:Lysine