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. 1978 Jul 1;173(1):11–17. doi: 10.1042/bj1730011

The reaction of Pseudomonas aeruginosa cytochrome c-551 oxidase with oxygen.

C Greenwood, D Barber, S R Parr, E Antonini, M Brunori, A Colosimo
PMCID: PMC1185743  PMID: 210764

Abstract

The reaction of ascorbate-reduced Pseudomonas cytochrome oxidase with oxygen was studied by using stopped-flow techniques at pH 7.0 and 25 degrees C. The observed time courses were complex, the reaction consisting of three phases. Of these, only the fastest process, with a second-order rate constant of 3.3 X 10(4) M-1.S-1, was dependent on oxygen concentration. The two slower processes were first-order reactions with rates of 1.0 +/- 0.4s-1 and 0.1 +/- 0.03s-1. A kinetic titration experiment revealed that the enzyme had a relatively low affinity constant for oxygen, approx. 10(4)M-1. Kinetic difference spectra were determined for all three reaction phases, showing each to have different characteristics. The fast-phase difference spectrum showed that changes occurred at both the haem c and haem d1 components of the enzyme during this process. These changes were consistent with the haem c becoming oxidized, but with the haem d1 assuming a form that did not correspond to the normal oxidized state, a situation that was not restored even after the second kinetic phase, which reflected further changes in the haem d1 component. The results are discussed in terms of a kinetic scheme.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Barber D., Parr S. R., Greenwood C. The reduction of Pseudomonas cytochrome c551 oxidase by chromous ions. Biochem J. 1977 Jun 1;163(3):629–632. doi: 10.1042/bj1630629. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. GIBSON Q. H., GREENWOOD C. THE REACTION OF CYTOCHROME OXIDASE WITH CYTOCHROME C. J Biol Chem. 1965 Feb;240:888–894. [PubMed] [Google Scholar]
  3. Gibson Q. H., Milnes L. Apparatus for rapid and sensitive spectrophotometry. Biochem J. 1964 Apr;91(1):161–171. doi: 10.1042/bj0910161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Greenwood C., Gibson Q. H. The reaction of reduced cytochrome C oxidase with oxygen. J Biol Chem. 1967 Apr 25;242(8):1782–1787. [PubMed] [Google Scholar]
  5. Greenwood C., Wilson M. T., Brunori M. Studies on partially reduced mammalian cytochrome oxidase. Reactions with carbon monoxide and oxygen. Biochem J. 1974 Feb;137(2):205–215. doi: 10.1042/bj1370205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. HORIO T., HIGASHI T., MATSUBARA H., KUSAI K., NAKAI M., OKUNUKI K. High purification and properties of Pseudomonas cytochrome oxidase. Biochim Biophys Acta. 1958 Aug;29(2):297–302. doi: 10.1016/0006-3002(58)90188-4. [DOI] [PubMed] [Google Scholar]
  7. HORIO T., HIGASHI T., YAMANAKA T., MATSUBARA H., OKUNUKI K. Purification and properties of cytochrome oxidase from Pseudomonas aeruginosa. J Biol Chem. 1961 Mar;236:944–951. [PubMed] [Google Scholar]
  8. Kuronen T., Ellfolk N. A new purification procedure and molecular properties of Pseudomonas cytochrome oxidase. Biochim Biophys Acta. 1972 Sep 20;275(3):308–318. doi: 10.1016/0005-2728(72)90212-5. [DOI] [PubMed] [Google Scholar]
  9. Parr S. R., Barber D., Greenwood C. A purification procedure for the soluble cytochrome oxidase and some other respiratory proteins from Pseudomonas aeruginosa. Biochem J. 1976 Aug 1;157(2):423–430. doi: 10.1042/bj1570423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Parr S. R., Barber D., Greenwood C., Brunori M. The electron-transfer reaction between azurin and the cytochrome c oxidase from Pseudomonas aeruginosa. Biochem J. 1977 Nov 1;167(2):447–455. doi: 10.1042/bj1670447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Parr S. R., Wilson M. T., Greenwood C. The reaction of Pseudomonas aeruginosa cytochrome c oxidase with carbon monoxide. Biochem J. 1975 Oct;151(1):51–59. doi: 10.1042/bj1510051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Shimada H., Orii Y. Oxidation-reduction behavior of the heme c and heme d moieties of Pseudomonas aeruginosa nitrite reductase and the formation of an oxygenated intermediate at heme d1. J Biochem. 1976 Jul;80(1):135–140. doi: 10.1093/oxfordjournals.jbchem.a131245. [DOI] [PubMed] [Google Scholar]
  13. Wharton D. C., Gibson Q. H. Cytochrome oxidase from Pseudomonas aeruginosa. IV. Reaction with oxygen and carbon monoxide. Biochim Biophys Acta. 1976 Jun 8;430(3):445–453. doi: 10.1016/0005-2728(76)90020-7. [DOI] [PubMed] [Google Scholar]
  14. YAMANAKA T., KIJIMOTO S., OKUNUKI K. Biological significance of Pseudomonas cytochrome oxidase in Pseudomonas aeruginosa. J Biochem. 1963 May;53:416–421. doi: 10.1093/oxfordjournals.jbchem.a127716. [DOI] [PubMed] [Google Scholar]
  15. YAMANAKA T., OKUNUKI K. Crystalline Pseudomonas cytochrome oxidase. II. Spectral properties of the enzyme. Biochim Biophys Acta. 1963 Mar 12;67:394–406. doi: 10.1016/0006-3002(63)91845-6. [DOI] [PubMed] [Google Scholar]
  16. YAMANAKA T., OTA A., OKUNUKI K. Activation of Pseudomonas cytochrome oxidase by catalase. J Biochem. 1961 May;49:414–420. doi: 10.1093/oxfordjournals.jbchem.a127319. [DOI] [PubMed] [Google Scholar]

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