Dependence of the 3-OH-Butyrate Dehydrogenase and Cytochrome c Oxidase Reactions on Intramitchondrial pH

  • Nancy L. Greenbaum
  • David F. Wilson


Suspensions of isolated rat liver mitochondria were incubated at constant extramitochondrial pH (pHe) with ATP, ADP, Pi, 3-OH-butyrate (3-OH-B), and acetoacetate (acac) (the last two were varied to maintain [3-OH-B]/[acac] constant), with or without sodium propionate to change the intramitochondrial pH. Measurements were made of the steady state water volume of the mitochondrial matrix, transmembrane pH difference, level of cytochrome c reduction, concentration of metabolites, and rate of oxygen consumption.

For each experiment, conditions were used for which PHi was near maximal and minimal values and the measured extramitochondrial [ATP], [ADP] and [Pi] were used to calculate log [ATP]/[ADP] [Pi]. When [3-OH-B]/[acac] and [cyt c +2]/[cyt c +3] were constant, and PHi was decreased from values near 7.7 to values near 7.2, log [ATP]/[ADP] [Pi] at high PHi was significantly (P<0.02) greater than at low pH. The mean values for change in log [ATP]/[ADP] [Pi] divided by change in pHi was 1.08 ± 0.15 (mean SEM). This agrees with the slope of 1.0 predicted if the energy available for ATP synthesis is dependent on the pH at which 3-OH-butyrate dehydrogenase operates, that is, the pH of the matrix space.

Plots of steady state respiratory rate vs. % cytochrome c reduction at different intra- and extramitochondrial pH values indicated that the rate of the cytochrome c oxidase reaction is dependent upon pHi and not on pHe. This implies that the matrix space is the source of protons consumed in the reduction of dioxygen to water in coupled mitochondria.


Respiratory Rate Oxidative Phosphorylation Free Energy Change Choline Chloride Matrix Space 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Holian, A. and Wilson, D.F. (1980) Biochemistry 19, 4213–4221CrossRefGoogle Scholar
  2. 2.
    Wilson, D.F. and Forman, N.G. (1982) Biochemistry 21, 1438–1444CrossRefGoogle Scholar
  3. 3.
    Greenbaum, N.L., and Wilson, D.F. (1985) J. Biol. Chem. 260, 873–879Google Scholar
  4. 4.
    Krebs, H.A., Mellany, J. and Williamson, D.H. (1962) Biochem. J. 82, 96–98Google Scholar
  5. 5.
    Williamson, D.H., Lund, P. and Krebs, H.A. (1967) Biochem. J. 103, 514–527Google Scholar
  6. 6.
    Erecinska, M., Veech, R.L., and Wilson, D.F. (1974) Arch. Biochem. Biophys. 160, 412–421CrossRefGoogle Scholar
  7. 7.
    Forman, N.G., and Wilson, D.F. (1982) J. Biol. Chem. 257, 12908–12915Google Scholar
  8. Wilson, D.F., Owen, C.S., and Holian, A. (1977)Arch. Biochem.Google Scholar
  9. Biophys. 182,749–762Google Scholar
  10. 9.
    Forman, N.G., and Wilson, D.F. (1983)J. Biol. Chem. 258, 8649–8655Google Scholar
  11. 10.
    Benzinger, T., Kitzinger, C., Hems, R., and Burton, K. (1959) Biochem. J. 11, 400–417Google Scholar
  12. 11.
    Rosing, J., and Slater, E.C. (1972) Biochim. Biophys. Acta. 267, 275–299CrossRefGoogle Scholar
  13. 12.
    Guynn, R., and Veech, R.L. (1973) J. Biol. Chem. 248, 6966–6969Google Scholar
  14. 13.
    Rodkey, F.L. and Ball, E.G. (1950) J. Biol. Chem. 183, 17–28Google Scholar
  15. 14.
    Dutton, P. L. Wilson, D. F. and Lee, C.P. (1971) Biochemistry, 9, 5077–5082CrossRefGoogle Scholar
  16. 15.
    Klingenberg, M. and Schollmeyer, P. (1961) Biochem. A. 335, 243–262Google Scholar
  17. 16.
    Chance, B. and Hollunger, G. (1961) J. Biol. Chem. 236, 1577–1584Google Scholar
  18. 17.
    Fleischer, S., Mclntyre, J.O., Churchill, P., Fleer, E., and Maurer, A., (1983) in: Structure and Function of Membrane Proteins (Quagliariello, E., and Palmieri, F. eds.) pp 283–300, Elsevier, AmsterdamGoogle Scholar
  19. 18.
    Maurer, A., Mclntyre, J.O., Churchill, S., and Fleischer, S. (1985) J. Biol. Chem. 260, 1661–1669Google Scholar
  20. 19.
    Mitchell, P., and Moyle, J. (1967) in: Biochemistry of Mitochondria (Slater, E.C., Kaniuga, Z., and Wojtczak, L. eds.) pp. 55–74, Academic Press, New YorkGoogle Scholar
  21. 20.
    Pietrobon, D., Zoratti, M., Azzone, G.F., Stucki, J.W., and Walz, D. (1982) Eur. J. Biochem. 127, 483–494CrossRefGoogle Scholar
  22. 21.
    Papa, S., Guerrieri, F., and Izzo, G. (1983) Biochem. J. 216, 259–272Google Scholar
  23. 22.
    Lehninger, A.L., Reynafarje, B., and Alexandre, A. (1978) in: Frontiers of Biological Energetics, Vol. I, (P.L. Dutton, J.S. Leigh, Jr. and A. Scarpa, eds.) pp. 384–393, Academic Press, New YorkGoogle Scholar
  24. 23.
    Ho, Y.-K., and Wang, J.H. (1981) J. Biol. Chem. 256, 2611–2614Google Scholar
  25. 24.
    Al-Shawi, M.K., and Brand, M.D. (1981) Biochem. J. 200, 539–546Google Scholar
  26. 25.
    Wikström, M., and Krab, K., (1979) Biochim. Biophys. Acta 549, 177–222Google Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Nancy L. Greenbaum
    • 1
  • David F. Wilson
    • 1
  1. 1.Department of Biochemistry and Biophysics School of MedicineUniversity of PennsylvaniaPhiladelphiaUSA

Personalised recommendations