title: Zn(2+) binding to the cytoplasmic side of Paracoccus denitrificans cytochrome c oxidase selectively uncouples electron transfer and proton translocation.
abstract: Using a combination of stopped-flow spectrophotometric proton pumping measurements and time-resolved potential measurements on black lipid membranes, we have investigated the effect of Zn(2+) ions on the proton transfer properties of Paracoccus denitrificans cytochrome c oxidase. When zinc was enclosed in the interior of cytochrome c oxidase containing liposomes, the H/e stoichiometry was found to gradually decrease with increasing Zn(2+) concentration. Half-inhibition of proton pumping was observed at [Zn(2+)](i)=75 microM corresponding to about 5-6 Zn(2+) ions per oxidase molecule. In addition, there was a significant increase in the respiratory control ratio of the proteoliposomes upon incorporation of Zn(2+). Time-resolved potential measurements on a black lipid membrane showed that the electrogenic phases slowed down in the presence of Zn(2+) correspond to phases that have been attributed to proton uptake from the cytoplasmic side and to proton pumping. We conclude that Zn(2+) ions bind close to or within the two proton transfer pathways of the bacterial cytochrome c oxidase.
authors.affiliation: Max Planck Institute of Biophysics, Department of Molecular Membrane Biology, Frankfurt am Main, Germany. kannt@mpibp-frankfurt.mpg.de
authors.forename: A
authors.initials: A
authors.lastname: Kannt
authors.forename: T
authors.initials: T
authors.lastname: Ostermann
authors.forename: H
authors.initials: H
authors.lastname: Müller
authors.forename: M
authors.initials: M
authors.lastname: Ruitenberg
mesh_terms: D001665:Binding Sites; D003593:Cytoplasm / Q000201:enzymology; D004579:Electron Transport; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D010231:Paracoccus denitrificans / Q000201:enzymology*; D011510:Proteolipids; D011522:Protons; D013053:Spectrophotometry; D055672:Static Electricity; D015032:Zinc / Q000378:metabolism*
chemical_list: D011510:Proteolipids; D011522:Protons; C020485:proteoliposomes; D003576:Electron Transport Complex IV; D015032:Zinc