title: The O(2) reduction and proton pumping gate mechanism of bovine heart cytochrome c oxidase.
abstract: X-ray structures of bovine heart cytochrome c oxidase with bound respiratory inhibitors (O(2) analogues) have been determined at 1.8-2.05Å resolution to investigate the function of the O(2) reduction site which includes two metal sites (Fe(a3)(2+) and Cu(B)(1+)). The X-ray structures of the CO- and NO-bound derivatives indicate that although there are three possible electron donors that can provide electrons to the bound O(2), located in the O(2) reduction site, the formation of the peroxide intermediate is effectively prevented to provide an O(2)-bound form as the initial intermediate. The structural change induced upon binding of CN(-) suggests a non-sequential 3-electron reduction of the bound O(2)(-) for the complete reduction without release of any reactive oxygen species. The X-ray structure of the derivative with CO bound to Cu(B)(1+) after photolysis from Fe(a3)(2+) demonstrates weak side-on binding. This suggests that Cu(B) controls the O(2) supply to Fe(a3)(2+) without electron transfer to provide sufficient time for collection of protons from the negative side of the mitochondrial membrane. The proton-pumping pathway of bovine heart cytochrome c oxidase includes a hydrogen-bond network and a water channel located in tandem between the positive and negative side of the mitochondrial membrane. Binding of a strong ligand to Fe(a3) induces a conformational change which significantly narrows the water channel and effectively blocks the back-leakage of protons from the hydrogen bond network. The proton pumping mechanism proposed by these X-ray structural analyses has been functionally confirmed by mutagenesis analyses of bovine heart cytochrome c oxidase. This article is part of a Special Issue entitled: Allosteric cooperativity in respiratory proteins.
authors: Department of Life Science, University of Hyogo, Hyogo, Japan. yoshi@sci.u-hyogo.ac.jp
authors: Shinya
authors: S
authors: Yoshikawa
authors: Kazumasa
authors: K
authors: Muramoto
authors: Kyoko
authors: K
authors: Shinzawa-Itoh
mesh_terms: D000818:Animals; D002417:Cattle; D003576:Electron Transport Complex IV / Q000737:chemistry / Q000378:metabolism*; D006860:Hydrogen Bonding; D009206:Myocardium / Q000201:enzymology; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D017494:Proton Pumps / Q000737:chemistry / Q000378:metabolism*; D011522:Protons
chemical_list: D017494:Proton Pumps; D011522:Protons; D003576:Electron Transport Complex IV; D010100:Oxygen