Electrochemical and spectroscopic investigations of the cytochrome bc1 complex from Rhodobacter capsulatus.
The cytochrome bc(1) complex from Rhodobacter capsulatus was investigated by protein electrochemistry and visible/IR spectroscopy. Infrared difference spectra, which represent redox-induced conformational changes of cofactors and their protein environments, show signals of the hemes, the quinone Q(i), and small conformational changes of the protein backbone. Furthermore, band features were tentatively assigned to protonated aspartic or glutamic acids involved in the redox transition of each of the b hemes, a proline in that of the [2Fe-2S] protein, and an arginine in that of cytochrome b(H). The midpoint potential of the [2Fe-2S] protein was determined for the first time at physiological temperature to be +290 mV at pH 8.7. The reduced minus oxidized difference extinction coefficients of the alpha-bands of the cytochromes were calculated as 11.5, 19, and 6.7 mM(-1) cm(-1) for cytochromes c(1), b(H), and b(L), respectively. A novel method has been developed to quantify protonation reactions of the complex during the redox reactions of its cofactors by evaluation of the buffer signals in the midinfrared region. Values will be discussed in relation to the pH dependence of the midpoint potentials.
Baymann
F
F
Institute de Biologie Physico-chimique, Paris, France. baymann@ibpc.fr
Robertson
D E
DE
Dutton
P L
PL
Mäntele
W
W
D000577:Amides; D004563:Electrochemistry; D004566:Electrodes; D014450:Electron Transport Complex III / Q000737:chemistry*; D006046:Gold; D006418:Heme / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007506:Iron-Sulfur Proteins / Q000737:chemistry; D046911:Macromolecular Substances; D010084:Oxidation-Reduction; D011489:Protein Denaturation; D011522:Protons; D011809:Quinones / Q000737:chemistry; D016354:Rhodobacter capsulatus / Q000201:enzymology*; D017550:Spectroscopy, Fourier Transform Infrared
D000577:Amides; D007506:Iron-Sulfur Proteins; D046911:Macromolecular Substances; D011522:Protons; D011809:Quinones; D006418:Heme; D006046:Gold; D014450:Electron Transport Complex III