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. 1983 Jan 15;210(1):199–205. doi: 10.1042/bj2100199

Protonmotive functions of cytochrome c oxidase in reconstituted vesicles. Influence of turnover rate on 'proton translocation'.

G Proteau, J M Wrigglesworth, P Nicholls
PMCID: PMC1154206  PMID: 6303310

Abstract

1. Oxidation of ferrocytochrome c by cytochrome c oxidase incorporated into proteoliposomes induces a transient acidification of the external medium. This change is dependent on the presence of valinomycin and can be abolished by carbonyl cyanide p-trifluoromethoxyphenylhydrazone or by nigericin. The H+/e- ratio for the initial acidification varies with the internal buffering capacity of the vesicles, and under suitable conditions approaches + 1, the pulse slowly decaying to give a net alkalinity change with H+/e- value approaching -1. 2. Inhibition of cytochrome c oxidase turnover by ferricytochrome c or by azide addition results in ferrocytochrome c-dependent H+ pulses with decreasing H+/e- ratios. The rate of the initial H+ production remains higher than the rate of equilibration of the pH gradient, indicating an intrinsic dependence of the H+/e- ratio on enzyme turnover. The final net alkalinity changes are relatively unaffected by turnover inhibition.

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Selected References

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  1. Brand M. D., Harper W. G., Nicholls D. G., Ingledew W. J. Unequal charge separation by different coupling spans of the mitochondrial electron transport chain. FEBS Lett. 1978 Nov 1;95(1):125–129. doi: 10.1016/0014-5793(78)80066-0. [DOI] [PubMed] [Google Scholar]
  2. CONRAD H., SMITH L. A study of the kinetics of the oxidation of cytochrome c by cytochrome c oxidase. Arch Biochem Biophys. 1956 Aug;63(2):403–413. doi: 10.1016/0003-9861(56)90055-8. [DOI] [PubMed] [Google Scholar]
  3. Casey R. P., Chappell J. B., Azzi A. Limited-turnover studies on proton translocation in reconstituted cytochrome c oxidase-containing vesicles. Biochem J. 1979 Jul 15;182(1):149–156. doi: 10.1042/bj1820149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Kornblatt J. A., Kells D. I., Williams G. R. The conformational states of oxidized and oxygenated beef-heart cytochrome c oxidase. Can J Biochem. 1975 Apr;53(4):461–466. doi: 10.1139/o75-063. [DOI] [PubMed] [Google Scholar]
  5. Krab K., Wikström M. Proton-translocating cytochrome c oxidase in artificial phospholipid vesicles. Biochim Biophys Acta. 1978 Oct 11;504(1):200–214. doi: 10.1016/0005-2728(78)90018-x. [DOI] [PubMed] [Google Scholar]
  6. Kuboyama M., Yong F. C., King T. E. Studies on cytochrome oxidase. 8. Preparation and some properties of cardiac cytochrome oxidase. J Biol Chem. 1972 Oct 25;247(20):6375–6383. [PubMed] [Google Scholar]
  7. Mitchell P., Moyle J. Acid-base titration across the membrane system of rat-liver mitochondria. Catalysis by uncouplers. Biochem J. 1967 Aug;104(2):588–600. doi: 10.1042/bj1040588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Mitchell P., Moyle J. Estimation of membrane potential and pH difference across the cristae membrane of rat liver mitochondria. Eur J Biochem. 1969 Feb;7(4):471–484. doi: 10.1111/j.1432-1033.1969.tb19633.x. [DOI] [PubMed] [Google Scholar]
  9. Mitchell P. The Ninth Sir Hans Krebs Lecture. Compartmentation and communication in living systems. Ligand conduction: a general catalytic principle in chemical, osmotic and chemiosmotic reaction systems. Eur J Biochem. 1979 Mar 15;95(1):1–20. doi: 10.1111/j.1432-1033.1979.tb12934.x. [DOI] [PubMed] [Google Scholar]
  10. Moyle J., Mitchell P. Cytochrome c oxidase is not a proton pump. FEBS Lett. 1978 Apr 15;88(2):268–272. doi: 10.1016/0014-5793(78)80190-2. [DOI] [PubMed] [Google Scholar]
  11. Nicholls D. G. The influence of respiration and ATP hydrolysis on the proton-electrochemical gradient across the inner membrane of rat-liver mitochondria as determined by ion distribution. Eur J Biochem. 1974 Dec 16;50(1):305–315. doi: 10.1111/j.1432-1033.1974.tb03899.x. [DOI] [PubMed] [Google Scholar]
  12. Nicholls P., Hildebrandt V. Binding of ligands and spectral shifts in cytochrome c oxidase. Biochem J. 1978 Jul 1;173(1):65–72. doi: 10.1042/bj1730065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Nicholls P., Hildebrandt V., Wrigglesworth J. M. Orientation and reactivity of cytochrome aa3 heme groups in proteoliposomes. Arch Biochem Biophys. 1980 Oct 15;204(2):533–543. doi: 10.1016/0003-9861(80)90065-x. [DOI] [PubMed] [Google Scholar]
  14. Papa S., Guerrieri F., Lorusso M., Izzo G., Boffoli D., Capuano F., Capitanio N., Altamura N. The H+/e- stoicheiometry of respiration-linked proton translocation in the cytochrome system of mitochondria. Biochem J. 1980 Oct 15;192(1):203–218. doi: 10.1042/bj1920203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Prochaska L. J., Bisson R., Capaldi R. A., Steffens G. C., Buse G. Inhibition of cytochrome c oxidase function by dicyclohexylcarbodiimide. Biochim Biophys Acta. 1981 Sep 14;637(2):360–373. doi: 10.1016/0005-2728(81)90175-4. [DOI] [PubMed] [Google Scholar]
  16. Racker E. A new procedure for the reconstitution of biologically active phospholipid vesicles. Biochem Biophys Res Commun. 1973 Nov 1;55(1):224–230. doi: 10.1016/s0006-291x(73)80083-x. [DOI] [PubMed] [Google Scholar]
  17. Reynafarje B., Brand M. D., Lehninger A. L. Evaluation of the H+/site ratio of mitochondrial electron transport from rate measurements. J Biol Chem. 1976 Dec 10;251(23):7442–7451. [PubMed] [Google Scholar]
  18. Sigel E., Carafoli E. Quantitative analysis of the proton and charge stoichiometry of cytochrome c oxidase from beef heart reconstituted into phospholipid vesicles. Eur J Biochem. 1980 Oct;111(2):299–306. doi: 10.1111/j.1432-1033.1980.tb04942.x. [DOI] [PubMed] [Google Scholar]
  19. Sigel E., Carafoli E. The proton pump of cytochrome c oxidase and its stoichiometry. Eur J Biochem. 1978 Aug 15;89(1):119–123. doi: 10.1111/j.1432-1033.1978.tb20903.x. [DOI] [PubMed] [Google Scholar]
  20. Solioz M., Carafoli E., Ludwig B. The cytochrome c oxidase of Paracoccus denitrificans pumps protons in a reconstituted system. J Biol Chem. 1982 Feb 25;257(4):1579–1582. [PubMed] [Google Scholar]
  21. Sorgato M. C., Ferguson S. J. Measurements of the components of the protonmotive force generated by cytochrome oxidase in submitochondrial particles. FEBS Lett. 1978 Jun 1;90(1):178–182. doi: 10.1016/0014-5793(78)80324-x. [DOI] [PubMed] [Google Scholar]
  22. Wikström M. K., Saari H. T. The mechanism of energy conservation and transduction by mitochondrial cytochrome c oxidase. Biochim Biophys Acta. 1977 Nov 17;462(2):347–361. doi: 10.1016/0005-2728(77)90133-5. [DOI] [PubMed] [Google Scholar]
  23. Wikström M., Krab K. Proton-pumping cytochrome c oxidase. Biochim Biophys Acta. 1979 Aug 17;549(2):177–122. doi: 10.1016/0304-4173(79)90014-4. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Wrigglesworth J. M., Nicholls P. Steady state spectra of cytochrome c oxidase in reconstituted vesicles. FEBS Lett. 1978 Jul 15;91(2):190–193. doi: 10.1016/0014-5793(78)81169-7. [DOI] [PubMed] [Google Scholar]
  26. Wrigglesworth J. M., Nicholls P. Turnover and vectorial properties of cytochrome c oxidase in reconstituted vesicles. Biochim Biophys Acta. 1979 Jul 10;547(1):36–46. doi: 10.1016/0005-2728(79)90093-8. [DOI] [PubMed] [Google Scholar]

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