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.
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
Brierley, G.P., Stoner, C.D. 1970. Swelling and contraction of heart mitochondria suspended in ammonium chloride.Biochemistry 9:708
Dryl, S. 1959. Antigenic transformation inParamecium aurelia after homologous antiserum treatment during autogamy and conjugation.J. Protozool. 6:S96
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
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
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
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
Hufnagel, L.A. 1967. Physical and Chemical Studies of Isolated Pellicles ofParamecium aurelia. Ph. D. Thesis, University of Pennsylvania, Philadelphia, Pennsylvania
Kagawa, Y., Racker, E. 1971. Partial resolution of the enzymes catalyzing oxidative phosphorylation.J. Biol. Chem. 246:5477
Knowles, A.F., Racker, E. 1975. Properties of a reconstituted calcium pump.J. Biol. Chem. 250:3538
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
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
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
Mueller, P., Rudin, D.O. 1968. Resting and action potentials in experimental bimolecular lipid membranes.J. Theoret. Biol. 18:222
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
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
Racker, E., Stoeckenius, W. 1974. Reconstitution of purple membrane vesicles catalyzing light-driven proton uptake and adenosine triphosphate formation.J. Biol. Chem. 249:662
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
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
Soldo, A.T., van Wagtendonk, W.J. 1969. The nutrition ofParamecium aurelia, Stock 299.J. Protozool. 16:500
Watson, M.R., Hopkins, J.M. 1962. Isolated cilia fromTetrahymena pyriformis.Exp. Cell Res. 28:280
Weiner, M.W. 1975. Mitochondrial permeability to chloride ion.Am. J. Physiol. 228:122
Author information
Authors and Affiliations
Rights 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
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01869662