Advertisement

Complete Tracking of Proton Flow in Thylakoids the Unit Conductance of CF0 is Greater than 10 fs

  • Gerald Schoenknecht
  • Holger Lill
  • Siegfried Engelbrecht
  • Wolfgang Junge
Part of the NATO ASI Series book series (NSSA, volume 133)

Abstract

We investigated the proton conductance of the channel portion of chlo-roplast ATP synthase (CF0) in thylakoids which were depleted from the soluble portion of the ATP synthase (CF1) by EDTA treatment. Proton pumps were stimulated by short flashes of light. Proton flux through CF0 was measured spectrophotometrically in three different ways: as proton efflux from the lumen (via neutral red), charge flow across the membrane (via electrochrom-ism) and proton influx into the medium (via phenol red). Hence we completely tracked the protons on their way from the lumen through CF0 into the medium.

A first treatment with EDTA removed up to 12% of total CF1 without increasing the proton permeability of the membranes. A 2nd EDTA treatment removed further 20% of CF1 and increased the proton permeability of membranes by 3 orders of magnitude. In control thylakoids a pH transient decayed with a relaxation time of 20–30 s. After the 2nd EDTA wash the decay time was 85 ms if the electric potential difference was shunted (e.g. by added gramicidin) and 7 ms if it was present. We calculated a lower limit of the protonic unit conductance of CF0 under the assumption that all exposed CF0 were proton conducting. It was 10 fS, corresponding to the passage of 6200 protons/s per CF0 (at 100 mV electric driving force) and by orders of magnitude higher than so far reported for any F0 channel.

Keywords

Electron Transport Chain Thylakoid Membrane Proton Conductance Proton Flux Electric Potential Difference 
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.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    McCarty, R.E. & Moroney, J.V. (1984) in: The Enzymes of Biological Membranes Vol. IV (ed. Martonosi, A.N.) pp. 383–413, Plenum Press, New YorkGoogle Scholar
  2. 2.
    Vignais, P.V. & Satre, M. (1984) Molec. and Cell. Biochem. 60, 33–70Google Scholar
  3. 3.
    Hennig, J. & Herrmann, R.G. (1986) Mol. Gen. Genet. 203, 117–128CrossRefGoogle Scholar
  4. 4.
    Cozens, A.L., Walker, J.E., Phillips, A.L., Huttly, A.K. & Gray, J.C. (1986) EMBO J. 5, 217–222PubMedGoogle Scholar
  5. 5.
    Sebald, W. & Hoppe, J. (1981) Curr. Top. Bioenerg. 12, 1–63Google Scholar
  6. 6.
    Fillingame, R.H. (1981) Curr. Top. Bioenerg. 11, 35–106Google Scholar
  7. 7.
    Hoppe, J. & Sebald, W. (1984) Biochim. Biophys. Acta 768, 1–27PubMedGoogle Scholar
  8. 8.
    Sone, N., Hamamoto, T. & Kagawa, Y. (1981) J. Biol. Chem. 256, 2873–2877PubMedGoogle Scholar
  9. 9.
    Berzbom, R.J., Mueller, D., Roos, P. & Andersson, B. (1981) in: Photosynthesis III. (ed. Akoyunoglou, G.) pp. 107–120 Balaban Int. Sci. Serv., Philadelphia, Pa.Google Scholar
  10. 10.
    Strotmann, H., Hesse, H. & Edelmann, K. (1973) Biochim. Biophys. Acta 314, 202–210PubMedCrossRefGoogle Scholar
  11. 11.
    Junge, W., Rumberg, B. & Schroeder, H. (1970) Eur. J. Biochem. 14, 575–581PubMedCrossRefGoogle Scholar
  12. 12.
    Davenport, J.W. & McCarty, R.E. (1981) J. Biol. Chem. 256, 8947–8954PubMedGoogle Scholar
  13. 13.
    Junge, W. & Jackson, J.B. (1982) in: Photosynthesis, Vol. 1 (ed. Govindjee) pp. 589–646, Academic, New YorkGoogle Scholar
  14. 14.
    Junge, W., Auslaender, W., McGeer, A. J. & Runge, T. (1979) Biochim. Biophys. Acta 546, 121–141PubMedCrossRefGoogle Scholar
  15. 15.
    Wille, B. & Lavergne, J. (1982) Photobiochem. Photobiophys. 4, 131–144Google Scholar
  16. 16.
    Auslaender, W. & Junge, W. (1975) FEBS Lett. 59, 310–315CrossRefGoogle Scholar
  17. 17.
    Hong, Y.Q. & Junge, W. (1983) Biochim. Biophys. Acta 722, 197–208CrossRefGoogle Scholar
  18. 18.
    DeWolf, F.A., Groen, B.H., VanHoute, L.P.A., Peters, F.A.L.J., Krab, K. & Kraayenhof, R. (1985) Biochim. Biophys. Acta 809, 204–214CrossRefGoogle Scholar
  19. 19.
    Junge, W., Schoenknecht, G. & Foerster, V. (1986) Biochim. Biophys. Acta -in press-Google Scholar
  20. 20.
    Junge, W. & Witt, H.T. (1968) Z. Naturf. 23b, 244–254Google Scholar
  21. 21.
    Junge, W. & Auslaender, W. (1973) Biochim. Biophys. Acta 333, 59–70Google Scholar
  22. 22.
    Polle, A. & Junge, W. (1986) Biochim. Biophys. Acta 848, 257–264CrossRefGoogle Scholar
  23. 23.
    Engelbrecht, S., Lill, H. & Junge, W. (1986) Eur. J. Biochem. -in press-Google Scholar
  24. 24.
    Sakurai, H., Shinohara, K., Hisabori, T. & Shinohara, K. (1981) J. Biochem. 90, 95–102PubMedGoogle Scholar
  25. 25.
    Sedmak, J.J. & Grossberg, S.E. (1977) Anal. Biochem. 79, 544–552PubMedCrossRefGoogle Scholar
  26. 26.
    Arnon, D.J. (1949) Plant Physiol. 24, 1–15PubMedCrossRefGoogle Scholar
  27. 27.
    Schmidt, G. & Graeber, P. (1985) Biochim. Biophys. Acta 808, 46–51CrossRefGoogle Scholar
  28. 28.
    Junge, W. (1976) in: Chemistry and Biochemistry of Plant Pigments, Vol. 2 (ed. Goodwin) pp. 233–333, Academic, New YorkGoogle Scholar
  29. 29.
    Foerster, V., Hong, Y.Q. & Junge, W. (1981) Biochim. Biophys. Acta 638, 141–152 Biochim. Biophys. Acta 807, 238–244CrossRefGoogle Scholar
  30. 36.
    Thomas, J.B., Minnaert, K. & Elbers, P.D. (1956) Acta Bot. Neerl. 5, 314–321Google Scholar
  31. 37.
    Schoenknecht, G., Junge, W., Lill, H. & Engelbrecht, S. (1986) FEBS Lett. 203, 289–294CrossRefGoogle Scholar
  32. 38.
    Lill, H., Engelbrecht, S., Schoenknecht, G. & Junge, W. (1986) Eur. J. Biochem. -in press-Google Scholar
  33. 39.
    Junge, W., Hong, Y.Q., Quian, L.P., & Viale, A. (1984) Proc. Natl. Acad. Sci. USA 81, 3078–3082PubMedCrossRefGoogle Scholar
  34. 40.
    Schmid, R., Shavit, N. & Junge, W. (1976) Biochim. Biophys. Acta 430, 145–153PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Gerald Schoenknecht
    • 1
  • Holger Lill
    • 1
  • Siegfried Engelbrecht
    • 1
  • Wolfgang Junge
    • 1
  1. 1.Biophysik, Fachbereich Biologie/ChemieUniversitaet OsnabrueckOsnabrueckGermany (FRG)

Personalised recommendations