Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Patch clamp study of the voltage-dependent anion channel in the thylakoid membrane

  • 87 Accesses

  • 27 Citations

Abstract

Measurements of single channel currents were performed on isolated membrane patches from osmotically swollen thylakoids of the Charophyte alga Nitellopsis obtusa. A channel with a high selectivity for anions over cations and a conductance of 100 to 110 pS (114 mM Cl−) was revealed. The channel has a bells-haped voltage-dependence of the open probability, with a maximum at about 0 mV. This dependence was explained by two gating processes, one causing channel closure at positive and one at negative potentials. The steepness of the voltage-dependence corresponded to approximately 2 elementary charges to be transferred across the entire membrane in each of the two gating processes. The analysis of the anion channel kinetics in the millisecond time domain revealed an e-fold increase of mean open and decrease of mean closed times when the membrane voltage was made more positive by 20 and 36 mV, respectively. Concert transitions of two identical anion channels between open and long inactivated states were observed, while the millisecond closed-open transitions of the two channels within a burst of activity were kinetically independent.

This is a preview of subscription content, log in to check access.

References

  1. Azzone, G., Benz, R., Bertl, A., Colombini, M., Crofts, A., Dilley, R., Dimroth, P., Dutton, P.L., Felle, H., Harold, F., Junge, W., Kaback, H.R., Knaff, D., Krulwich, T., Lodish, H., Malmström, B., Maloney, P., Mannella, C., Padan, E., Papa, S., Rottenberg, H., Rudnick, G., Rydström, J., Silverstein, T., Skulachev, V., Slayman, C., Tedeschi, H., Wikström, M., Wilson, T.H. 1993. Transmembrane measurements across bioenergetic membranes. Biochim. Biophys. Acta 1183:1–3

  2. Blatz, A.L., Magleby, K.L. 1986. Quantitative description of three modes of activity of fast chloride channels from rat sceletal muscle. J. Physiol. 378:141–174

  3. Bulychev, A.A., Andrianov, V.K., Kurella, G.A., Litvin, F.F. 1972. Microelectrode measurements of the transmembrane potential of chloroplasts and its photoinduced changes. Nature 236:175–177

  4. Bulychev, A.A., Antonov, V.F., Schevchenko, E.V. 1992. Patch-clamp studies of light-induced currents across the thylakoid membrane of isolated chloroplasts. Biochim. Biophys. Acta 1099:16–24

  5. Bulychev, A.A., Vredenberg, W.J. 1976. Effect of ionophores A23187 and nigericin on the light-induced redistribution of Mg2+, K+, and H+ across the thylakoid membrane. Biochim. Biophys. Acta 423:548–556

  6. Colombini, M. 1989. Voltage gating in the mitochondrial channel, VDAC. J. Membrane Biol. 111:103–111

  7. Deamer, D.W., Packer, L. 1969. Light-dependent anion transport in isolated spinach chloroplasts. Biochim. Biophys. Acta 172:539–545

  8. Dilley, R.A., Vernon, L.P. 1965. Ion and water transport processes related to the light-dependent shrinkage of spinach chloroplasts. Arch. Biochem. Biophys. 111:365–375

  9. Draber, S., Hansen, U.P. 1994. Fast single-channel measurements resolve the blocking effect of Cs+ on the K+ channel. Biophys. J. 67:120–129

  10. Enz, C., Steinkamp, T., Wagner, R. 1993. Ion channels in the thylakoid membrane (a patch-clamp study). Biochim. Biophys. Acta 1143:67–76

  11. Franciolini, F., Adams, D.J. 1994. Functional properties of background chloride channels. In: Handbook of Membrane Channels. Molecular and Cellular Physiology. C. Peracchia, editor, pp. 255–266. Academic Press, New York

  12. Geletyuk, V.I., Kazachenko, V.N. 1985. Single Cl− channels in molluscan neurons: Multiplicity of the conductance states. J. Membrane Biol. 86:9–15

  13. Hamill, O.P., Marty, A., Nehere, E., Sakmann, B., Sigworth, F.J. 1981. Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches. Pflüegers Arch. 391:85–100

  14. Hind, G., Nakatani, H.Y., Izawa, S. 1974. Light-dependent redistribution of ions in suspensions of chloroplast thylakoid membranes. Proc. Natl. Acad. Sci. USA 71:1484–1488

  15. Junge, W., Jackson, J.B. 1982. The development of electrochemical potential gradients across photosynthetic membranes. In: Photosynthesis: Energy Conversion by Plants and Bacteria. Govindjee, editor. Vol. I, pp. 589–646. Academic Press, New York

  16. Miller, C. 1982. Open-state substructure of single chloride channels from Torpedo electroplax. Phil. Trans. R. Soc. Lond. B 299:401–411

  17. Morier, N., Sauve, R. 1994. Analysis of a novel double-barreled anion channel from rat liver rough endoplasmic reticulum. Biophys. J. 67:590–602

  18. Muniz, J., Pottosin, I.I., Sandoval, L. 1995. Patch-clamp study of vascular plant chloroplast: single channels and photocurrents. J. Bioenerg. Biomembr. (in press)

  19. Pottosin, I.I. 1992. Single channel recording in the chloroplast envelope. FEBS Lett. 308:87–90

  20. Pottosin, I.I., Schönknecht, G. 1995. Anion and cation channels in the thylakoid membrane. In: Proceedings of the Xth International Photosynthesis Congress. P. Mathis, editor. Kluwer Academic Publisher, Netherlands (in press)

  21. Remiš, D., Bulychev, A.A., Kurella, G.A. 1986. The electrical and chemical components of the protonmotive force in chloroplasts as measured with capillary and pH-sensitive microelectrodes. Biochim. Biophys. Acta 852:68–73

  22. Rottenberg, H., Grunwald, T., Avron, M. 1972. Determination of ΔpH in chloroplasts 1. Distribution of [14C]methylamine. Eur. J. Biochem. 25:54–63

  23. Schönknecht, G., Althoff, G., Apley, E., Wagner, R., Junge, W. 1989. Cation channels by subunit III of the channel portion of the chloroplast H+-ATPase. FEBS Lett. 258:190–194

  24. Schönknecht, G., Hedrich, R., Junge, W., Raschke, K. 1988. A voltage dependent Chloride channel in the photosynthetic membrane of a higher plant. Nature 336:589–592

  25. Schröppel-Meier, G., Kaiser, W.M. 1988. Ion homeostasis in chloroplasts under salinity and mineral deficiency I. Solute concentrations in leaves and chloroplasts from spinach plants under NaCl or NaNO3 salinity. Plant Physiol. 87:822–827

  26. Sorgato, M.C., Keller, B.U., Stühmer, W. 1987. Patch-clamping of the inner mitochondrial membrane reveals a voltage-dependent ion channel. Nature 330:498–500

  27. Sorgato, M.C., Moran, O. 1993. Channels in mitochondrial membranes: knowns, unknowns, and prospects for the future. Critical Rev. Biochem. Mol. Biol. 18:127–171

  28. Speer, M., Kaiser, W.M. 1991. Ion relations of symplastic and apoplastic space in leaves from Spinacia oleracea L. and Pisum sativum L. under salinity. Plant Physiol. 97:990–997

  29. Thaler, M., Simonis, W., Schönknecht, G. 1992. Light-dependent changes of the cytoplasmic H+ and Cl activity in the green alga Eremosphaera viridis. Plant Physiol. 99:103–110

  30. Tester, M., Blatt, M.R. 1989. Direct measurement of K+ channels in thylakoid membranes by incorporation of vesicles into planar lipid bilayers. Plant Physiol. 91:249–252

  31. Vambutas, V., Schechter, S. 1983. Chloride ion transport and its inhibition in thylakoid membranes. Arch. Biochem. Biophys. 224:442–448

  32. White, M.M., Miller, C. 1979. A voltage-dependent anion channel from the electric organ of Torpedo californica. J. Biol. Chem. 254: 10161–10166

Download references

Author information

Correspondence to I. I. Pottosin.

Additional information

This work was financially supported by the Deutsche Forschungsgemeinschaft (SFB 176 TP B11) and by a research fellowship from the Alexander von Humboldt Foundation to I.I. Pottosin.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Pottosin, I.I., Schönknecht, G. Patch clamp study of the voltage-dependent anion channel in the thylakoid membrane. J. Membarin Biol. 148, 143–156 (1995). https://doi.org/10.1007/BF00207270

Download citation

Key words

  • Anion channel
  • Channel gating
  • Patch clamp
  • Thylakoid membrane
  • Nitellopsis obtusa