Redox-induced transitions in bovine cytochrome bc1 complex studied by perfusion-induced ATR-FTIR spectroscopy.
Redox transitions in a film of detergent-purified bovine cytochrome bc(1) complex were investigated by perfusion-induced attenuated total reflection Fourier transform infrared (ATR-FTIR) spectroscopy. The technique provides a flexible method for generating redox-induced IR changes of components of bovine cytochrome bc(1) complex at a high signal:noise ratio. These IR redox difference spectra arise from perturbations of prosthetic groups and surrounding protein. Visible difference spectra were recorded synchronously using a light beam reflected from the exposed prism surface and provided a quantitative means of determining the redox transitions that were occurring. IR and visible redox difference spectra of iron-sulfur protein/cytochrome c(1), heme b(H), and heme b(L) were separated by selective reduction and/or oxidation that extends published data on the homologous bacterial enzyme. Several bands could be tentatively assigned to redox-sensitive modes of hemes and ubiquinone and changes in the surrounding protein by comparison with available data for bacterial bc(1) complex, other related heme proteins, and model compounds. Some tentative assignments of further signals to specific amino acids are made on the basis of known crystal structures.
D000818:Animals; D002417:Cattle; D018360:Crystallography, X-Ray; D003575:Cytochromes c1 / Q000737:chemistry; D014450:Electron Transport Complex III / Q000737:chemistry*; D006418:Heme / Q000031:analogs & derivatives; D007506:Iron-Sulfur Proteins / Q000737:chemistry; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010477:Perfusion; D012680:Sensitivity and Specificity; D013053:Spectrophotometry / Q000379:methods; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation / Q000379:methods*; D014451:Ubiquinone / Q000737:chemistry
D007506:Iron-Sulfur Proteins; D014451:Ubiquinone; D006418:Heme; D003575:Cytochromes c1; D014450:Electron Transport Complex III