Skip to main content
Log in

Reconstitution in planar lipid bilayers of a voltage-dependent anion-selective channel obtained from paramecium mitochondria

  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Summary

We have incorporated into planar lipid bilayer membranes a voltage-dependent, anion-selective channel (VDAC) obtained fromParamecium aurelia. VDAC-containing membranes have the following properties: (1) The steady-state conductance of a many-channel membrane is maximal when the transmembrane potential is zero and decreases as a steep function of both positive and negative voltage. (2) The fraction of time that an individual channel stays open is strongly voltage dependent in a manner that parallels the voltage dependence of a many-channel membrane. (3) The conductance of the open channel is about 500 pmho in 0.1 to 1.0m salt solutions and is ohmic. (4) The channel is about 7 times more permeable to Cl than to K+ and is impermeable to Ca++. The procedure for obtaining VDAC and the properties of the channel are highly reproducible.

VDAC activity was found, upon fractionation of the paramecium membranes, to come from the mitochondria. We note that the published data on mitochondrial Cl permeability suggest that there may indeed be a voltage-dependent Cl permeability in mitochondria.

The method of incorporating VDAC into planar lipid bilayers may be generally useful for reconstituting biological transport systems in these membranes.

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

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Brierley, G.P. 1970. Energy-linked alteration of the permeability of heart mitochondria to chloride and other anions.Biochemistry 9:697

    Google Scholar 

  • Brierley, G.P., Stoner, C.D. 1970. Swelling and contraction of heart mitochondria suspended in ammonium chloride.Biochemistry 9:708

    Google Scholar 

  • Dryl, S. 1959. Antigenic transformation inParamecium aurelia after homologous antiserum treatment during autogamy and conjugation.J. Protozool. 6:S96

    Google Scholar 

  • Ehrenstein, G., Lecar, H., Nossal, R. 1970. The nature of the negative resistance in bimolecular lipid membranes containing excitability-inducing material.J. Gen. Physiol. 55:119

    Google Scholar 

  • Goldin, S., Tong, S.W. 1974. Reconstitution of active transport catalyzed by the purified sodium and potassium ion-stimulated adenosine triphosphatase from canine and renal medulla.J. Biol. Chem. 249:5907

    Google Scholar 

  • Hilden, S., Rhee, H.M., Hokin, L.E. 1974. Sodium transport by phospholipid vesicles containing purified sodium and potassium ion-activated adenosine triphosphatase.J. Biol. Chem. 249:7432

    Google Scholar 

  • Hinkle, P.C., Kim, J.J., Racker, E. 1972. Ion transport and respiratory control in vesicles formed from cytochrome oxidase and phospholipids.J. Biol. Chem. 247:1338

    Google Scholar 

  • Hufnagel, L.A. 1967. Physical and Chemical Studies of Isolated Pellicles ofParamecium aurelia. Ph. D. Thesis, University of Pennsylvania, Philadelphia, Pennsylvania

    Google Scholar 

  • Kagawa, Y., Racker, E. 1971. Partial resolution of the enzymes catalyzing oxidative phosphorylation.J. Biol. Chem. 246:5477

    Google Scholar 

  • Knowles, A.F., Racker, E. 1975. Properties of a reconstituted calcium pump.J. Biol. Chem. 250:3538

    Google Scholar 

  • Lowry, O.H., Rosenbrough, N.J., Farr, A.L., Randall, R.J. 1951. Protein measurement with the folin phenol reagent.J. Biol. Chem. 193:265

    Google Scholar 

  • Montal, M. 1974a. Formation of bimolecular membranes from lipid monolayers.In: Methods in Enzymology. Vol. 32 Part B, p. 545. S. Fleischer and L. Packer, editors. Academic Press, New York

    Google Scholar 

  • Montal, M. 1974b. Lipid-protein assembly and the reconstitution of biological membranes.In: Perspectives in Membrane Biology. p. 591. S. Estrada-O and C. Gitler, editors. Academic Press, New York

    Google Scholar 

  • Mueller, P., Rudin, D.O. 1968. Resting and action potentials in experimental bimolecular lipid membranes.J. Theoret. Biol. 18:222

    Google Scholar 

  • Mueller, P., Rudin, D.O., Tien, H.Ti, Wescott, W.C. 1963. Methods for the formation of single bimolecular lipid membranes in aqueous solution.J. Phys. Chem. 67:534

    Google Scholar 

  • Nicholls, D.G. 1974. Hamster brown-adipose-tissue mitochondria. The chloride permeability of the inner membrane under respiring conditions, the influence of purine nucleotides.Eur. J. Biochem. 49:585

    Google Scholar 

  • Racker, E., Stoeckenius, W. 1974. Reconstitution of purple membrane vesicles catalyzing light-driven proton uptake and adenosine triphosphate formation.J. Biol. Chem. 249:662

    Google Scholar 

  • Semenza, G. 1974. The transport systems for sugars in the small intestine. Reconstitution of one of them.In: Drugs and Transport Processes. B.A. Callingham, editor. p. 317. Macmillan Press, London

    Google Scholar 

  • Soldo, A.T., Godoy, G.A., van Wagtendonk, W.J. 1966. Growth of particle-bearing and particle-freeParamecium aurelia in axenic culture.J. Protozool. 13:492

    Google Scholar 

  • Soldo, A.T., van Wagtendonk, W.J. 1969. The nutrition ofParamecium aurelia, Stock 299.J. Protozool. 16:500

    Google Scholar 

  • Watson, M.R., Hopkins, J.M. 1962. Isolated cilia fromTetrahymena pyriformis.Exp. Cell Res. 28:280

    Google Scholar 

  • Weiner, M.W. 1975. Mitochondrial permeability to chloride ion.Am. J. Physiol. 228:122

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Schein, S.J., Colombini, M. & Finkelstein, A. Reconstitution in planar lipid bilayers of a voltage-dependent anion-selective channel obtained from paramecium mitochondria. J. Membrain Biol. 30, 99–120 (1976). https://doi.org/10.1007/BF01869662

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01869662

Keywords

Navigation