Theoretical and computational analysis of the membrane potential generated by cytochrome c oxidase upon single electron injection into the enzyme.
We have developed theory and the computational scheme for the analysis of the kinetics of the membrane potential generated by cytochrome c oxidase upon single electron injection into the enzyme. The theory allows one to connect the charge motions inside the enzyme to the membrane potential observed in the experiments by using data from the "dielectric topography" map of the enzyme that we have created. The developed theory is applied for the analysis of the potentiometric data recently reported by the Wikström group [I. Belevich, D.A. Bloch, N. Belevich, M. Wikström and M.I. Verkhovsky, Exploring the proton pump mechanism of cytochrome c oxidase in real time, Proc. Natl. Acad. Sci. U. S. A. 104 (2007) 2685-2690] on the O to E transition in Paracoccus denitrificans oxidase. Our analysis suggests, that the electron transfer to the binuclear center is coupled to a proton transfer (proton loading) to a group just "above" the binuclear center of the enzyme, from which the pumped proton is subsequently expelled by the chemical proton arriving to the binuclear center. The identity of the pump site could not be determined with certainty, but could be localized to the group of residues His326 (His291 in bovine), propionates of heme a(3), Arg 473/474, and Trp164. The analysis also suggests that the dielectric distance from the P-side to Fe a is 0.4 or larger. The difficulties and pitfalls of quantitative interpretation of potentiometric data are discussed.
Department of Chemistry, University of California, Davis, CA 95616, USA.
Ryogo
R
Sugitani
Emile S
ES
Medvedev
Alexei A
AA
Stuchebrukhov
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D019295:Computational Biology; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000378:metabolism*; D004583:Electrons; D007700:Kinetics; D008564:Membrane Potentials; D008954:Models, Biological; D050505:Mutant Proteins / Q000378:metabolism; D010231:Paracoccus denitrificans / Q000166:cytology / Q000201:enzymology* / Q000502:physiology*; D017433:Protein Structure, Secondary; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011522:Protons; D012242:Rhodobacter sphaeroides / Q000378:metabolism
D050505:Mutant Proteins; D021122:Protein Subunits; D011522:Protons; D003576:Electron Transport Complex IV