title: The reaction of Pseudomonas aeruginosa cytochrome c oxidase with carbon monoxide.
abstract: The binding of CO to ascorbate-reduced Pseudomonas cytochrome oxidase was investigated by static-titration, stopped-flow and flash-photolytic techniques. Static-titration data indicated that the binding process was non-stoicheiometric, with a Hill number of 1.44. Stopped-flow kinetics obtained on the binding of CO to reduced Pseudomonas cytochrome oxidase were biphasic in form; the faster rate exhibited a linear dependence on CO concentration with a second-order rate constant of 2 X 10(4) M-1-s-1, whereas the slower reaction rapidly reached a pseudo-first-order rate limit at approx. 1s-1. The relative proportions of the two phases observed in stopped-flow experiments also showed a dependency on CO concentration, the slower phase increasing as the CO concentration decreased. The kinetics of CO recombination after flash-photolytic dissociation of the reduced Pseudomonas cytochrome oxidase-CO complex were also biphasic in character, both phases showing a linear pseudo-first-order rate dependence on CO concentration. The second-order rate constants were determined as 3.6 X 10(4)M-1-s-1 and 1.6 X 10(4)M-1-s-1 respectively. Again the relative proportions of the two phases varied with CO concentration, the slower phase predominating at low CO concentrations. CO dissociation from the enzyme-CO complex measured in the presence of O2 and NO indicated the presence of two rates, of the order of 0.03s-1 and 0.15s-1. When sodium dithionite was used as a reducing agent for the Pseudomonas cytochrome oxidase, the CO-combination kinetics observed by both stopped flow and flash photolysis were extremely complex and not able to be simply analysed.
mesh_terms: D001665:Binding Sites; D002248:Carbon Monoxide / Q000378:metabolism*; D003201:Computers; D003576:Electron Transport Complex IV / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D009589:Nitrogen Oxides; D010100:Oxygen; D010782:Photolysis; D011550:Pseudomonas aeruginosa / Q000201:enzymology*
chemical_list: D008024:Ligands; D009589:Nitrogen Oxides; D002248:Carbon Monoxide; D003576:Electron Transport Complex IV; D010100:Oxygen