title: Studies of the primary oxygen intermediate in the reaction of fully reduced cytochrome oxidase.
abstract: The formation and disappearance of a photosensitive species during the reaction of reduced cytochrome c oxidase (putatively a3II.O2), EC 1.9.3.1, has been followed by (a) mixing a3II.CO with O2 in a stopped flow apparatus; (b) initiating the oxygen-oxidase reaction by removing CO with a laser flash; (c) probing the reaction mixture for photosensitivity with a second laser flash. Photosensitivity appears in the reaction mixture after the first laser flash, reaches a maximum after 50-60 microseconds ([O2] greater than 100 microM), and disappears in a further 50-100 microseconds. The kinetics can be represented by the scheme [formula: see text]. In species B, O2 is associated with the protein, possibly CuB, but not with the heme. Species C is the photosensitive a3II.O2 complex, and in D, a3 iron has been oxidized. The formation of species C is responsible for the rapid phase of absorbance change in the oxidase-oxygen reaction. The rate of reaction with oxygen approaches the limit of 35,000 s-1 at high oxygen. Nitric oxide, however, reacts with FeII oxidase with a rate of 1 x 10(8) M-1 s-1, which is accurately maintained up to an observed rate of 10(5) s-1. In flash photolysis experiments, approximately half of the photodissociated nitric oxidase recombines in a biphasic geminate reaction with rates of 1 x 10(8) s-1 and 1 x 10(7) s-1.
authors.affiliation: Department of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853.
authors.forename: R S
authors.initials: RS
authors.lastname: Blackmore
authors.forename: C
authors.initials: C
authors.lastname: Greenwood
authors.forename: Q H
authors.initials: QH
authors.lastname: Gibson
mesh_terms: D000818:Animals; D002417:Cattle; D003576:Electron Transport Complex IV / Q000737:chemistry*; D007700:Kinetics; D009206:Myocardium / Q000201:enzymology; D009569:Nitric Oxide / Q000737:chemistry; D010084:Oxidation-Reduction; D010100:Oxygen / Q000737:chemistry*; D010777:Photochemistry
chemical_list: D009569:Nitric Oxide; D003576:Electron Transport Complex IV; D010100:Oxygen