Abstract
Addition of Triton X-100 to planar bilayers composed of dioleoyl phosphatidyl choline, diphytanoyl phosphatidyl choline or mono-oleoyl glycerol induces single channel-like events when electrical conductivity across the bilayer is measured. Addition of divalent cations or protons causes channels to disappear; single channel conductance of remaining channels is not significantly altered; addition of EDTA or alkali (respectively) reverses the effect. It is concluded that sensitivity to divalent cations and protons need not be dependent on specific channel proteins or pore-forming toxins, but may be a feature of any aqueous pore across a lipid milieu.
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Akutsu, H., Seelig, J. 1981. Interaction of metal ions with phosphatidylcholine bilayer membranes. Biochemistry 20:7366–7373
Alder, G.M., Arnold, W.M., Bashford, C.L., Drake, A.F., Pasternak, C.A., Zimmerman, U. 1991. Divalent cation-sensitive pores formed by natural and synthetic melittin and by triton X-100. Biochim. Biophys. Acta 1061:111–120
Alonso, A., Goni, F.M. 1983. Effect of detergents and fusogenic lipids on phospholipid phase transitions. J. Membrane Biol. 71:183–187
Antonov, V.F., Petrov, V.V., Molnar, A.A., Prevoditelev, D.A., Ivanov, A.S. 1980. The appearance of single-ion channels in unmodified lipid bilayer membranes at the phase transition temperature. Nature 283:585–587
Avigad, L.S., Bernheimer, A.W. 1976. Inhibition by zinc of hemolysis induced by bacterial and other cytolytic agents. Infect. Immun. 13:1378–1381
Bashford, C.L., Alder, G.M., Menestrina, G., Micklem, K.J., Murphy, J.J., Pasternak, C.A. 1986. Membrane damage by hemolytic viruses, toxins, complement and other cytotoxic agents: a common mechanism blocked by divalent cations. J. Biol. Chem. 261: 9300–9308
Bashford, C.L., Alder, G.M., Patel, K., Pasternak, C.A. 1984. Common action of certain viruses, toxins, and activated complement: pore formation and its prevention by extracellular Ca2+. Biosci. Rep. 4:797–805
Bashford, C.L., Alder, G.M., Graham, J.M., Menestrina, G., Pasternak, C.A. 1988a. Ion modulation of membrane permeability: Effect of cations on intact cells and on cells and phospholipid bilayers treated with pore-forming agents. J. Membrane Biol. 103:79–94
Bashford, C.L., Menestrina, G., Henkart, P.A., Pasternak, C.A. 1988b. Cell damage by cytolysin. Spontaneous recovery and reversible inhibition by divalent cations. J. Immunol. 141:3965–3974
Boyle M.D., Langone J.J., Borsos T. 1979. Studies on the terminal stages of immune hemolysis. IV. Effect of metal salts. J. Immunol. 122:1209–1213
Burnet, F.M. 1949. Haemolysis by Newcastle disease virus. Nature 164:1008
Gilly, W.F., Armstrong, C.M. 1982a. Slowing of sodium channel opening kinetics in squid axon by extracellular zinc. J. Gen. Physiol. 79:935–964
Gilly, W.F., Armstrong, C.M. 1982b. Divalent cations and the activation kinetics of potassium channels in squid giant axons. J. Gen. Physiol 79:965–966
Gotlib, V.A., Lebedeva, N.E., Stakhov, S.V., Rostovtseva, T.K., Lev, A.A. 1992. Lateral phase separation in lipid bilayers in the presence of non-ionic detergent triton X-100. Membrane Biol. 9:629–674 (in Russian)
Gotze, O., Haupt, I., Fisher, H. 1968. Immune haemolysis: reaction of the terminal complement component. Nature 217:1165–1167
Hille, B. 1992. Ionic Channels of Excitable Membranes. Second edition. Sinauer Associates, Sunderland, MA
Henkart, P.A. 1985. Mechanism of lymphocyte-mediated cytotoxicity. Annu. Rev. Immunol. 3:31–58
Korchev, Y.E., Bashford, C.L., Pasternak, C.A. 1992. Differential sensitivity of pneumolysin-induced channels to gating by divalent cations. J. Membrane Biol. 127:195–203
Lacapere, J.-J., Stokes, D.L., Chatenay, D. 1992. Atomic force microscopy of three-dimensional membrane protein crystals. Biophys. J. 63:303–308
Lee, C.Y., McCammon, J.A., Rossky, P.J. 1984. The structure of liquid water at an extended hydrophobic surface. J. Chem. Phys. 80:4448–4455
Lev, A.A., Korchev, Y.E., Rostovtseva, T.K., Bashford, C.L., Pasternak, C.A. 1992. Lipid impregnated nuclear filters as a new model for studies of surface conductance and single channel phenomena. In: Biophysics of Membrane Transport, Proceedings of Eleventh School J. Kuczera, and S. Przestalski, editors. pp. 321–349. Agricultural University of Wroclaw, Poland
Lev, A.A., Korchev, Y.E., Rostovtseva, T.K., Bashford, C.L., Edmonds, D.T., Pasternak, C.A. 1993. Rapid switching of ion current in narrow pores: implications for biological ion channels. Proc. R. Soc. Lond. B 252:187–192
Levitt, D.G. 1988. Solvation effects on the transport of ions across cell membranes. In: The Chemical Physics of Solvation, Part C, Solvation Phenomena in Specific Physical, Chemical, and Biological Systems R.R., Dogonadze, E. Kalman, A.A. Kornishev, and J. Ulstrup, editors. pp. 741–750. Elsevier, Amsterdam
Liu, J., Blumenthal, K.M. 1988. Membrane damage by Cerebratulus lacteus cytolysin A-III. Effects of monovalent and divalent cations on A-III hemolytic activity. Biochim. Biophys. Acta 937:153–160
Madigan, S.T., Whitbread, J.A., Katz, E.R. 1990. A Dictyostelium discoideum mutant exhibiting calcium dependent, high level detergent resistance. J. Bact. 172:2785–2787
Maeda, T., Ohnishi, S.-I. 1980. Activation of influenza virus by acidic media causes hemolysis and fusion of erythrocytes. FEBS Lett. 122:283–287
McLaughlin, A., Grathwohl, C., McLaughlin, S. 1978. The adsorption of divalent cations to phosphatidylcholine bilayer membranes. Biochim. Biophys. Acta 513:338–357
Menestrina, G. 1986. Ionic channels formed by Staphylococcus aureus α-toxin: Voltage-dependent inhibition by divalent and trivalent cations. J. Membrane Biol. 90:177–190
Menestrina, G., Bashford, C.L., Pasternak, C.A. 1990. Pore-forming toxins: experiments with S. aureus α-toxin, C. perfringens Θ-toxin and E. coli haemolysin in lipid bilayers, liposomes and intact cells. Toxicon 28:477–491
Micklem, K.J., Alder, G.M., Buckley, G.D., Murphy, J., Pasternak, C.A. 1988. Protection against complement-mediated cell damage by Ca2+ and Zn2+. Complement 5:141–152
Mironov, S.L., Sokolov Y.V., Chanturiya, A.N., Lishko, V.K. 1986. Channels produced by spider venoms in bilayer lipid membrane: mechanisms of ion transport and toxic action. Biochim. Biophys. Acta 862:185–198
Nachshen, D.A. 1984. Selectivity of the Ca binding site in synaptosome Ca channels. Inhibition of Ca influx by multivalent metal cations. J. Gen. Physiol. 83:941–967
Nagel, W., Natochin, Y., Crabbe, J. 1988. Effects of divalent cations in chloride movement across amphibian skin. Pfluegers. Arch. 411:540–545
Obaid, A.L., Socolar, S.J., Rose, B. 1983. Cell-to-cell channels with two independently regulated gates in series: Analysis of junctional conductance modulation by membrane potential, calcium, and pH. J. Membrane Biol 73:69–89
Pasternak, C.A. 1991. Effect of divalent cations on membranes: reversible and irreversible closure of channels induced by membrane-inserting proteins and other agents. Romanian J. Biophys. 1:3–11
Pasternak, C.A., Alder, G.M., Bashford, C.L., Korchev, Y.E., Pederzolli, C., Rostovtseva, T.K. 1992. Membrane damage: common mechanisms of induction and prevention. FEMS Microbiol. Immunol. 105:83–92
Pasternak, C.A., Bashford, C.L., Korchev, Y.E., Rostovtseva, T.K., Lev, A.A. 1993. Modulation of surface flow by divalent cations and protons. Colloids & Surfaces 77:119–124
Pasternak, C.A., Micklem, K.J. 1974. The biochemistry of virus-induced cell fusion. Changes in membrane integrity. Biochem. J. 140:405–411
Patel, K., Pasternak, C.A. 1985. Permeability changes elicited by influenza and Sendai virus: separation of fusion and leakage by pH jump experiments. J. Gen. Virol. 66:767–775
Pietrobon, D., Prod'hom, B., Hess, P. 1988. Conformational changes associated with ion permeation in L-type calcium channels. Nature 333:373–376
Prod'hom, B., Pietrobon, D., Hess, P. 1987. Direct measurement of proton transfer rates to a group controlling the dihydropyridine-sensitive Ca2+ channel. Nature 329:243–246
Rose, B., Loewenstein, W.R. 1975. Permeability of cell junction depends on local cytoplasmic calcium activity. Nature 254:250–252
Rostovtseva, T.K., Lev, A.A. 1986. Lipid peroxidation induced single ion channels in the lipid bilayer membranes. Symposium on Activated Oxygen Species in Biological Systems, Varna. p. 81
Rostovtseva, T.K., Osipov, V.V., Lev, A.A. 1987. Dependence of single gramicidin channel conductance on potential induced by adsorbtion of 1-anilino-8 naphthalene sulfonate anions on lipid membranes. Biol. Membranes 4:955–964 (in Russian)
Sandvig, K., Olsnes, S. 1980. Diphtheria toxin entry into cells is facilitated by low pH. J. Cell Biol. 87:828–832
Schindler, H. 1980. Formation of planar bilayers from artificial or native membrane vesicles. FEBS Lett. 122:77–79
Schlieper, P., de Robertis, E. 1977. Triton X-100 as a channel-forming substance in artificial lipid bilayer membranes. Arch. Biochem. Biophys. 184:204–208
Smart, T.G. 1990. Uncultured lobster muscle, cultured neurons and brain slices: the neurophysiology of zinc. J. Pharm. Pharmacol. 42:377–387
Tanaka, J.C., Furman, R.E., Barchi, R.L. 1986. Skeletal muscle sodium channels. In: Ion Channel Reconstitution C. Miller, editor. pp. 277–305, Plenum, New York
Tatulian, S.A. 1983. Effect of lipid phase transition on the binding of anions to dimyristoylphosphatidylcholine liposomes. Biochim. Biophys. Acta 736:189–195
Thelestam, M., Mollby, R. 1980. Interaction of streptolysin O from Streptococcus pyogenes and theta toxin from Clostridium perfringens with human fibroblasts. Infect. Immunol. 29:863–872
Van Zutphen, Merola, A.J., Brierley, G.P., Cornwall, D.G. 1972. The interaction of non-ionic detergents with lipid bilayer membranes. Arch. Biochem. Biophys. 152:755–766
Westbrook, G.L., Mayer, M.I. 1987. Micromolar concentrations of Zn2+ antagonise NMDA and GABA responses in hippocampal neurons. Nature 328:640–643
Wilmsen, H.U., Pattus, F., Buckley, J.T. 1990. Aerolysin, a hemolysin from Aeromonas hydrophila, forms voltage-gated channels in planar lipid bilayers. J. Membrane Biol. 115:71–81
Woll, K.H., Leibowitz, M.D., Neumcke, B., Hille, B. 1987. A high conductance anion channel in adult amphibian skeletal muscle. Pfluegers Arch. 410:632–640
Wolosin, J.M., Forte, J.G. 1985. K+ Cl− conductances in the apical membrane from secreting oxyntic cells are concurrently inhibited by divalent cations. J. Membrane Biol. 83:261–272
Woodbury, D.J. 1989. Pure lipid vesicles can induce channel-like conductances in planar bilayers. J. Membrane Biol. 109:145–150
Woodhull, A.M. 1973. Ionic blockage of sodium channels in nerve. J. Gen. Physiol. 61:687–708
Yoshikawa, K., Sakabe, K., Matsubara, Y., Ota, T. 1988. Self-excitation in a porous medium doped with sorbitan monooleate (SPAN-80) induced by a Na+/K+ concentration gradient. Biophys. Chem. 21:33–39
Young, J.D.-E., Nathan, C.F., Podack, E.R., Palladino, M.A., Cohn, Z.A. 1986. Functional channel formation associated with cytotoxic T-cell granules. Proc. Natl. Acad. Sci. USA 83:150–154
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We are grateful to Dr. D.T. Edmonds and Prof. R.J.P. Williams for critical discussion, to Glenn Alder for technical assistance, to Ms. B. Bashford and Ms. S.G. Pelc for preparing the paper, and to the Cell Surface Research Fund, the Royal Society (A.A.L.), UNESCO (Molecular and Cell Biology Network) and The Wellcome Trust for financial support.
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Rostovtseva, T.K., Bashford, C.L., Lev, A.A. et al. Triton channels are sensitive to divalent cations and protons. J. Membarin Biol. 141, 83–90 (1994). https://doi.org/10.1007/BF00232876
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DOI: https://doi.org/10.1007/BF00232876