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. 1995 Sep 1;310(Pt 2):477–481. doi: 10.1042/bj3100477

The nature of mitochondrial respiration and discrimination between membrane and pump properties.

M Canton 1, S Luvisetto 1, I Schmehl 1, G F Azzone 1
PMCID: PMC1135920  PMID: 7654185

Abstract

A new criterion is utilized for the interpretation of flow-force relationships in rat liver mitochondria. The criterion is based on the view that the nature of the relationship between the H+/O ratio and the membrane potential can be inferred from the relationship between ohmic-uncoupler-induced extra respiration and the membrane potential. Thus a linear relationship between extra respiration and membrane potential indicates unequivocally the independence of the H+/O ratio from the membrane potential and the leak nature of the resting respiration [Brand, Chien, and Diolez (1994) Biochem. J. 297, 27-29]. On the other hand, a non-linear relationship indicates that the H+/O ratio is dependent on the membrane potential. The experimental assessment of this relationship in the presence of an additional ohmic leak, however, is rendered difficult by both the uncoupler-induced depression of membrane potential and the limited range of dependence of the H+/O ratio on the membrane potential. We have selected conditions, i.e. incubation of mitochondria at low temperatures, where the extent of non-linearity is markedly increased. It appears that the nature of the resting respiration of mitochondria in vitro is markedly dependent on the temperature: at low temperatures the percentage of resting respiration due to membrane leak decreases and that due to intrinsic uncoupling of the proton pumps increases.

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Selected References

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  1. Berman M. C. Energy coupling and uncoupling of active calcium transport by sarcoplasmic reticulum membranes. Biochim Biophys Acta. 1982 Aug 11;694(1):95–121. doi: 10.1016/0304-4157(82)90015-6. [DOI] [PubMed] [Google Scholar]
  2. Brand M. D., Chien L. F., Diolez P. Experimental discrimination between proton leak and redox slip during mitochondrial electron transport. Biochem J. 1994 Jan 1;297(Pt 1):27–29. doi: 10.1042/bj2970027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brand M. D., Steverding D., Kadenbach B., Stevenson P. M., Hafner R. P. The mechanism of the increase in mitochondrial proton permeability induced by thyroid hormones. Eur J Biochem. 1992 Jun 15;206(3):775–781. doi: 10.1111/j.1432-1033.1992.tb16984.x. [DOI] [PubMed] [Google Scholar]
  4. Brown G. C., Brand M. D. Changes in permeability to protons and other cations at high proton motive force in rat liver mitochondria. Biochem J. 1986 Feb 15;234(1):75–81. doi: 10.1042/bj2340075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brown G. C., Brand M. D. On the nature of the mitochondrial proton leak. Biochim Biophys Acta. 1991 Aug 2;1059(1):55–62. doi: 10.1016/s0005-2728(05)80187-2. [DOI] [PubMed] [Google Scholar]
  6. Casey R. P., Thelen M., Azzi A. Dicyclohexylcarbodiimide binds specifically and covalently to cytochrome c oxidase while inhibiting its H+-translocating activity. J Biol Chem. 1980 May 10;255(9):3994–4000. [PubMed] [Google Scholar]
  7. Garlid K. D., Beavis A. D., Ratkje S. K. On the nature of ion leaks in energy-transducing membranes. Biochim Biophys Acta. 1989 Sep 28;976(2-3):109–120. doi: 10.1016/s0005-2728(89)80219-1. [DOI] [PubMed] [Google Scholar]
  8. Hafner R. P., Nobes C. D., McGown A. D., Brand M. D. Altered relationship between protonmotive force and respiration rate in non-phosphorylating liver mitochondria isolated from rats of different thyroid hormone status. Eur J Biochem. 1988 Dec 15;178(2):511–518. doi: 10.1111/j.1432-1033.1988.tb14477.x. [DOI] [PubMed] [Google Scholar]
  9. Inesi G., de Meis L. Regulation of steady state filling in sarcoplasmic reticulum. Roles of back-inhibition, leakage, and slippage of the calcium pump. J Biol Chem. 1989 Apr 5;264(10):5929–5936. [PubMed] [Google Scholar]
  10. Kell D. B., John P., Ferguson S. J. On the current-voltage relationships of energy-transducing membranes: phosphorylating membrane vesicles from Paracoccus denitrificans [proceedings]. Biochem Soc Trans. 1978;6(6):1292–1295. doi: 10.1042/bst0061292. [DOI] [PubMed] [Google Scholar]
  11. Krishnamoorthy G., Hinkle P. C. Non-ohmic proton conductance of mitochondria and liposomes. Biochemistry. 1984 Apr 10;23(8):1640–1645. doi: 10.1021/bi00303a009. [DOI] [PubMed] [Google Scholar]
  12. Li P. M., Morgan J. E., Nilsson T., Ma M., Chan S. I. Heat treatment of cytochrome c oxidase perturbs the CuA site and affects proton pumping behavior. Biochemistry. 1988 Sep 20;27(19):7538–7546. doi: 10.1021/bi00419a054. [DOI] [PubMed] [Google Scholar]
  13. Luvisetto S., Conti E., Buso M., Azzone G. F. Flux ratios and pump stoichiometries at sites II and III in liver mitochondria. Effect of slips and leaks. J Biol Chem. 1991 Jan 15;266(2):1034–1042. [PubMed] [Google Scholar]
  14. Luvisetto S., Pietrobon D., Azzone G. F. Uncoupling of oxidative phosphorylation. 1. Protonophoric effects account only partially for uncoupling. Biochemistry. 1987 Nov 17;26(23):7332–7338. doi: 10.1021/bi00397a021. [DOI] [PubMed] [Google Scholar]
  15. Luvisetto S., Schmehl I., Intravaia E., Conti E., Azzone G. F. Mechanism of loss of thermodynamic control in mitochondria due to hyperthyroidism and temperature. J Biol Chem. 1992 Aug 5;267(22):15348–15355. [PubMed] [Google Scholar]
  16. Nicholls D. G. The effective proton conductance of the inner membrane of mitochondria from brown adipose tissue. Dependency on proton electrochemical potential gradient. Eur J Biochem. 1977 Jul 15;77(2):349–356. doi: 10.1111/j.1432-1033.1977.tb11674.x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. O'Shea P. S., Petrone G., Casey R. P., Azzi A. The current-voltage relationships of liposomes and mitochondria. Biochem J. 1984 May 1;219(3):719–726. doi: 10.1042/bj2190719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Ouhabi R., Rigoulet M., Lavie J. L., Guérin B. Respiration in non-phosphorylating yeast mitochondria. Roles of non-ohmic proton conductance and intrinsic uncoupling. Biochim Biophys Acta. 1991 Nov 7;1060(3):293–298. doi: 10.1016/s0005-2728(05)80319-6. [DOI] [PubMed] [Google Scholar]
  20. Pietrobon D., Azzone G. F., Walz D. Effect of funiculosin and antimycin A on the redox-driven H+-pumps in mitochondria: on the nature of "leaks'. Eur J Biochem. 1981 Jul;117(2):389–394. doi: 10.1111/j.1432-1033.1981.tb06350.x. [DOI] [PubMed] [Google Scholar]
  21. Pietrobon D., Caplan S. R. Flow-force relationships for a six-state proton pump model: intrinsic uncoupling, kinetic equivalence of input and output forces, and domain of approximate linearity. Biochemistry. 1985 Oct 8;24(21):5764–5776. doi: 10.1021/bi00342a012. [DOI] [PubMed] [Google Scholar]
  22. Pietrobon D., Luvisetto S., Azzone G. F. Uncoupling of oxidative phosphorylation. 2. Alternative mechanisms: intrinsic uncoupling or decoupling? Biochemistry. 1987 Nov 17;26(23):7339–7347. doi: 10.1021/bi00397a022. [DOI] [PubMed] [Google Scholar]
  23. Pietrobon D., Zoratti M., Azzone G. F., Caplan S. R. Intrinsic uncoupling of mitochondrial proton pumps. 2. Modeling studies. Biochemistry. 1986 Feb 25;25(4):767–775. doi: 10.1021/bi00352a005. [DOI] [PubMed] [Google Scholar]
  24. Pietrobon D., Zoratti M., Azzone G. F., Stucki J. W., Walz D. Non-equilibrium thermodynamic assessment of redox-driven H+ pumps in mitochondria. Eur J Biochem. 1982 Oct;127(3):483–494. doi: 10.1111/j.1432-1033.1982.tb06897.x. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Rottenberg H. Decoupling of oxidative phosphorylation and photophosphorylation. Biochim Biophys Acta. 1990 Jul 17;1018(1):1–17. doi: 10.1016/0005-2728(90)90103-b. [DOI] [PubMed] [Google Scholar]
  27. Rottenberg H., Hashimoto K. Fatty acid uncoupling of oxidative phosphorylation in rat liver mitochondria. Biochemistry. 1986 Apr 8;25(7):1747–1755. doi: 10.1021/bi00355a045. [DOI] [PubMed] [Google Scholar]
  28. Sone N., Nicholls P. Effect of heat treatment on oxidase activity and proton-pumping capability of proteoliposome-incorporated beef heart cytochrome aa3. Biochemistry. 1984 Dec 18;23(26):6550–6554. doi: 10.1021/bi00321a042. [DOI] [PubMed] [Google Scholar]
  29. Steverding D., Kadenbach B. Influence of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline modification on proton translocation and membrane potential of reconstituted cytochrome-c oxidase support "proton slippage". J Biol Chem. 1991 May 5;266(13):8097–8101. [PubMed] [Google Scholar]
  30. Steverding D., Köhnke D., Ludwig B., Kadenbach B. Proton slippage in cytochrome c oxidase of Paracoccus denitrificans. Membrane-potential measurements with the two-subunit and three-subunit enzyme. Eur J Biochem. 1993 Mar 15;212(3):827–831. doi: 10.1111/j.1432-1033.1993.tb17724.x. [DOI] [PubMed] [Google Scholar]
  31. Wrigglesworth J. M., Cooper C. E., Sharpe M. A., Nicholls P. The proteoliposomal steady state. Effect of size, capacitance and membrane permeability on cytochrome-oxidase-induced ion gradients. Biochem J. 1990 Aug 15;270(1):109–118. doi: 10.1042/bj2700109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Zoratti M., Favaron M., Pietrobon D., Azzone G. F. Intrinsic uncoupling of mitochondrial proton pumps. 1. Non-ohmic conductance cannot account for the nonlinear dependence of static head respiration on delta microH. Biochemistry. 1986 Feb 25;25(4):760–767. doi: 10.1021/bi00352a004. [DOI] [PubMed] [Google Scholar]
  33. van Dam K., Shinohara Y., Unami A., Yoshida K., Terada H. Slipping pumps or proton leaks in oxidative phosphorylation. The local anesthetic bupivacaine causes slip in cytochrome c oxidase of mitochondria. FEBS Lett. 1990 Dec 17;277(1-2):131–133. doi: 10.1016/0014-5793(90)80826-5. [DOI] [PubMed] [Google Scholar]

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