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. 1986 Mar 15;234(3):569–572. doi: 10.1042/bj2340569

Transient kinetics of subunit-III-depleted cytochrome c oxidase.

F Malatesta, G Antonini, P Sarti, M Brunori
PMCID: PMC1146609  PMID: 3013160

Abstract

Cytochrome c oxidase from ox heart was depleted of subunit III and its transient kinetic properties studied by stopped-flow and flash photolysis. It was found that the overall mechanism of electron transfer is very similar for subunit-III-depleted and native oxidase, although significant differences in some kinetic parameters have been detected. These include the second-order rate constant for cytochrome c oxidation and the rate-limiting step of the overall process. Moreover, at low cytochrome c/oxidase ratios (where the number of reducing equivalents is insufficient), the rate of reoxidation of cytochrome a was found to be very slow, even in air, and in fact for the subunit-III-depleted enzyme is even slower than for the native oxidase. The stability of reduced cytochrome a excludes the likelihood that removal of subunit III leads to a new O2-binding site, and the result may be relevant to the lowered vectorial H+/e- stoichiometry. The subunit-III-depleted oxidase can be pulsed under appropriate conditions and its combination with CO is unchanged, as shown by kinetic experiments and difference spectroscopy.

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Selected References

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

  1. Antonini E., Brunori M., Colosimo A., Greenwood C., Wilson M. T. Oxygen "pulsed" cytochrome c oxidase: functional properties and catalytic relevance. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3128–3132. doi: 10.1073/pnas.74.8.3128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Antonini E., Brunori M., Greenwood C., Malmström B. G. Catalytic mechanism of cytochrome oxidase. Nature. 1970 Dec 5;228(5275):936–937. doi: 10.1038/228936a0. [DOI] [PubMed] [Google Scholar]
  3. Antonini G., Brunori M., Colosimo A., Malatesta F., Sarti P. Pulsed cytochrome c oxidase. J Inorg Biochem. 1985 Mar-Apr;23(3-4):289–293. doi: 10.1016/0162-0134(85)85037-6. [DOI] [PubMed] [Google Scholar]
  4. Azzi A. Cytochrome c oxidase. Towards a clarification of its structure, interactions and mechanism. Biochim Biophys Acta. 1980 Dec;594(4):231–252. doi: 10.1016/0304-4173(80)90002-6. [DOI] [PubMed] [Google Scholar]
  5. Brunori M., Antonini G., Colosimo A., Malatesta F., Sarti P., Jones M. G., Wilson M. T. Stopped-flow studies of cytochrome oxidase reconstituted into liposomes: proton pumping and control of activity. J Inorg Biochem. 1985 Mar-Apr;23(3-4):373–379. doi: 10.1016/0162-0134(85)85048-0. [DOI] [PubMed] [Google Scholar]
  6. Brunori M., Giacometti G. M. Photochemistry of hemoproteins. Methods Enzymol. 1981;76:582–595. doi: 10.1016/0076-6879(81)76146-9. [DOI] [PubMed] [Google Scholar]
  7. Buse G., Meinecke L., Bruch B. The protein formula of beef heart cytochrome c oxidase. J Inorg Biochem. 1985 Mar-Apr;23(3-4):149–153. doi: 10.1016/0162-0134(85)85019-4. [DOI] [PubMed] [Google Scholar]
  8. Capaldi R. A., Malatesta F., Darley-Usmar V. M. Structure of cytochrome c oxidase. Biochim Biophys Acta. 1983 Jul 15;726(2):135–148. doi: 10.1016/0304-4173(83)90003-4. [DOI] [PubMed] [Google Scholar]
  9. GIBSON Q. H., GREENWOOD C. Reactions of cytochrome oxidase with oxygen and carbon monoxide. Biochem J. 1963 Mar;86:541–554. doi: 10.1042/bj0860541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Greenwood C., Brittain T. Studies on partially reduced mammalian cytochrome oxidase reactions with ferrocytochrome c. Biochem J. 1976 Sep 1;157(3):591–598. doi: 10.1042/bj1570591. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mayevsky A., Jamieson D., Chance B. Oxygen poisoning in the unanesthetized brain: correlation of the oxidation-reduction state of pyridine nucleotide with electrical activity. Brain Res. 1974 Aug 23;76(3):481–491. doi: 10.1016/0006-8993(74)90824-5. [DOI] [PubMed] [Google Scholar]
  12. Nałeçz K. A., Bolli R., Ludwig B., Azzi A. The role of subunit III in bovine cytochrome c oxidase. Comparison between native, subunit III-depleted and Paracoccus denitrificans enzymes. Biochim Biophys Acta. 1985 Jul 17;808(2):259–272. doi: 10.1016/0005-2728(85)90008-8. [DOI] [PubMed] [Google Scholar]
  13. Penttilä T. Properties and reconstitution of a cytochrome oxidase deficient in subunit III. Eur J Biochem. 1983 Jun 15;133(2):355–361. doi: 10.1111/j.1432-1033.1983.tb07470.x. [DOI] [PubMed] [Google Scholar]
  14. Puettner I., Carafoli E., Malatesta F. Spectroscopic and functional properties of subunit III-depleted cytochrome oxidase. J Biol Chem. 1985 Mar 25;260(6):3719–3723. [PubMed] [Google Scholar]
  15. Sarti P., Jones M. G., Antonini G., Malatesta F., Colosimo A., Wilson M. T., Brunori M. Kinetics of redox-linked proton pumping activity of native and subunit III-depleted cytochrome c oxidase: a stopped-flow investigation. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4876–4880. doi: 10.1073/pnas.82.15.4876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Thompson D. A., Ferguson-Miller S. Lipid and subunit III depleted cytochrome c oxidase purified by horse cytochrome c affinity chromatography in lauryl maltoside. Biochemistry. 1983 Jun 21;22(13):3178–3187. doi: 10.1021/bi00282a022. [DOI] [PubMed] [Google Scholar]
  17. Thompson D. A., Gregory L., Ferguson-Miller S. Cytochrome c oxidase depleted of subunit III: proton-pumping, respiratory control, and pH dependence of the midpoint potential of cytochrome a. J Inorg Biochem. 1985 Mar-Apr;23(3-4):357–364. doi: 10.1016/0162-0134(85)85046-7. [DOI] [PubMed] [Google Scholar]
  18. Vanneste W. H. The stoichiometry and absorption spectra of components a and a-3 in cytochrome c oxidase. Biochemistry. 1966 Mar;5(3):838–848. doi: 10.1021/bi00867a005. [DOI] [PubMed] [Google Scholar]
  19. Wilson M. T., Greenwood C., Brunori M., Antonini E. Kinetic studies on the reaction between cytochrome c oxidase and ferrocytochrome c. Biochem J. 1975 Apr;147(1):145–153. doi: 10.1042/bj1470145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Wilson M. T., Peterson J., Antonini E., Brunori M., Colosimo A., Wyman J. A plausible two-state model for cytochrome c oxidase. Proc Natl Acad Sci U S A. 1981 Nov;78(11):7115–7118. doi: 10.1073/pnas.78.11.7115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. YONETANI T. Studies on cytochrome oxidase. III. Improved preparation and some properties. J Biol Chem. 1961 Jun;236:1680–1688. [PubMed] [Google Scholar]

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