Elevated proton leak of the intermediate OH in cytochrome c oxidase.
The kinetics of the formation and relaxation of transmembrane electric potential (Deltapsi) during the complete single turnover of CcO was studied in the bovine heart mitochondrial and the aa(3)-type Paracoccus denitrificans enzymes incorporated into proteoliposome membrane. The real-time Deltapsi kinetics was followed by the direct electrometry technique. The prompt oxidation of CcO and formation of the activated, oxidized (O(H)) state of the enzyme leaves the enzyme trapped in the open state that provides an internal leak for protons and thus facilitates dissipation of Deltapsi (tau(app) < or = 0.5-0.8 s). By contrast, when the enzyme in the O(H) state is rapidly re-reduced by sequential electron delivery, Deltapsi dissipates much slower (tau(app) > 3 s). In P. denitrificans CcO proteoliposomes the accelerated Deltapsi dissipation is slowed down by a mutational block of the proton conductance through the D-, but not K-channel. We concluded that in contrast to the other intermediates the O(H) state of CcO is vulnerable to the elevated internal proton leak that proceeds via the D-channel.
Bloch
Dmitry A
DA
Institute of Biotechnology, 00014 University of Helsinki, Helsinki, Finland.
Jasaitis
Audrius
A
Verkhovsky
Michael I
MI
D000465:Algorithms; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D002417:Cattle; D003576:Electron Transport Complex IV / Q000737:chemistry* / Q000235:genetics; D004583:Electrons; D007700:Kinetics; D053078:Membrane Potential, Mitochondrial; D008567:Membranes, Artificial; D008929:Mitochondria, Heart / Q000737:chemistry; D016297:Mutagenesis, Site-Directed; D010084:Oxidation-Reduction; D010231:Paracoccus denitrificans; D011510:Proteolipids / Q000737:chemistry; D011522:Protons
D001426:Bacterial Proteins; D008567:Membranes, Artificial; D011510:Proteolipids; D011522:Protons; C020485:proteoliposomes; D003576:Electron Transport Complex IV