Monoclonal antibody to human cytochrome c: effect on electron-transfer reactions.
A monoclonal antibody has been produced to an antigenic site on human cytochrome c which includes amino acid number 58 (isoleucine). This area is on the bottom back of the cytochrome, removed from the postulated binding/reaction sites for oxidase and reductase, but in the area of the molecule where an appreciable change in conformation is seen on oxidation-reduction. In spectrophotometric assays, where binding of cytochrome c to the oxidase or reductase is rate-limiting, the antibody gave stimulation of the reductase reaction under some conditions, where the oxidase reaction was inhibited. Also variation of the pH of the reaction medium resulted in differential effects on the oxidase and reductase reactions. Different effects of the antibody were seen when the oxidase was assayed polarographically, as compared to the spectrophotometric measurements. The data show that the binding/reaction sites on cytochrome c for the oxidase and reductase must be different. They suggest that binding of antibody may affect conformational changes in the whole molecule, distorting the binding/reaction sites. Conformational changes may be involved as a control mechanism in cytochrome c-mediated electron-transfer reactions.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000918:Antibody Specificity; D001665:Binding Sites; D001666:Binding Sites, Antibody; D002417:Cattle; D003574:Cytochrome c Group / Q000276:immunology* / Q000378:metabolism; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000378:metabolism; D006801:Humans; D009245:NADH Dehydrogenase / Q000378:metabolism; D010084:Oxidation-Reduction; D011048:Polarography; D011487:Protein Conformation; D013053:Spectrophotometry
D000911:Antibodies, Monoclonal; D003574:Cytochrome c Group; D009245:NADH Dehydrogenase; D003576:Electron Transport Complex IV