Summary
The compound, 4,5,6,7-tetrachloro-2-methylbenzimidazole (TMB), has been found to markedly modify the steady-state valinomycin-mediated conductance of potassium (K+) ions through lipid bilayer membranes. TMB alone does not contribute significantly to membrane conductance, being electrically neutral in solution. In one of two classes of experiments (I), valinomycin is first added to the aqueous phases then changes of membrane conductance accompanying stepwise addition of TMB to the water are measured. In a second class of experiments (II), valinomycin is added to the membrane-forming solution, follwed by TMB additions to the surrounding water. In both cases membrane conductance shows an initial increase with increasing TMB concentration which is more pronounced at lower K+ ion concentration. At TMB concentrations in excess of 10−5 m, membrane conductance becomes independent of K+ ion concentration, in contrast to the linear dependence observed at TMB concentrations below 10−7 m. This transition is accompanied by a change of high field current-voltage characteristics from superlinear (or weakly sublinear) to a strongly sublinear form. All of these observations may be correlated by the kinetic model for carriermedicated transport proposed by Läuger and Stark (Biochim. Biophys. Acta 211:458, 1970) from which it may be concluded that valinomycin-mediated ion transport is limited by back diffusion of the uncomplexed carrier at high TMB concentrations. Experiments of class I reveal a sharp drop of conductance at high (>10−5 m) TMB concentration, not seen in class II experiments, which is attributed to blocked entry of uncomplexed carrier from the aqueous phases. Valinomycin initially in the membrane is removed by lateral diffusion to the surrounding torus. The time dependence of this removal has been studied in a separate series of experiments, leading to a measured coefficient of lateral diffusion for valinomycin of 5×10−6 cm2/sec at 25°C. This value is about two orders of magnitude larger than the corresponding coefficient for transmembrane carrier diffusion, and provides further evidence for localization of valinomycin in the membrane/solution interfaces.
Similar content being viewed by others
References
Andreoli, T.E., Tieffenberg, M., Tosteson, D.C. 1967. The effect of valinomycin on the ionic permeability of thin lipid membranes.J. Gen. Physiol. 50:2527
Benz, R., Stark, G. 1975. Kinetics of macrotetralide-induced ion transport across lipid bilayer membranes.Biochim. Biophys. Acta 382:27
Benz, R., Stark, G., Janko, K., Läuger, P. 1973. Valinomycin-mediated ion transport through neutral lipid membranes: Influence of hydrocarbon chain length and temperature.J. Membrane Biol. 14:339
Ciani, S.M., Eisenman, G., Laprade, R., Szabo, G. 1972. Theoretical analysis of carriermediated electrical properties of bilayer membranes. In: Membranes — A Series of Advances. G. Eisenman, Editor. Vol. 2, p. 61. Marcel Dekker, New York
Ciani, S., Eisenman, G., Szabo, G. 1969. A theory for the effects of neutral carriers such as the macrotetralide actin antibiotics on the electric properties of bilayer membranes.J. Membrane Biol. 1:1
Cogan, U., Shinitzky, M., Weber, G., Nishida, T. 1973. Microviscosity and order in the hydrocar region of phospholipid and phospholipid-cholesterol dispersions determined with fluorescent probes.Biochemistry 12:521
Eisenman, G., Krasne, S., Ciani, S. 1975. The kinetic and equilibrium components of selective ionic permeability mediated by nactin-and valinomycin-type carriers having systematically varied degrees of methylation.Ann. N. Y. Acad. Sci (In press)
Hall, J.E., Mead, C.A., Szabo, G. 1973. A barrier model for current flow in lipid bilayer membranes.J. Membrane Biol. 11:75
Hladky, S.B. 1972. The steady-state theory of the carrier transport of ions.J. Membrane Biol. 10:67
Hladky, S.B. 1973. The effect of stirring on the flux of carriers into black lipid membranes.Biochim. Biophys. Acta. 307:261
Hladky, S.B. 1974. The energy barriers to ion transport by nonactin across thin lipid membranes.Biochim. Biophys. Acta 352:71
Hladky, S.B. 1975. Tests of the carrier model for ion transport by nonactin and trinactin.Biochim. Biophys. Acta 375:327
Hsu, M., Chan, I.S. 1973. Nuclear magnetic resonance studies of the interaction of valinomycin with unsonicated lecithin bilayers.Biochemistry 12:3872
Huebner, J.S., Bruner, L.J. 1972. Apparatus for measurement of the dynamic current-voltage characteristics of membranes.J. Phys. E., Sci. Instrum. 5:310
Jost, W. 1960. Diffusion in Solids, Liquids, Gases. (Third printing with addendum.) Academic Press, Inc., New York, p. 45.
Knoll, W., Stark, G. 1975. An extended kinetic analysis of valinomycin induced Rb-transport through monoglyceride membranes.J. Membrane Biol. 25:249
Kuo, K.-H., Bruner, L.J. 1973. Uncoupler antagonism of valinomycin induced bilayer membrane conductance.Biochem. Biophys. Res. Commun. 52:1079
Kuo, K.-H., Fukuto, T.R., Miller, T.A., Bruner, L.J. 1976. Blocking of valinomycin-mediated bilayer membrane conductance by substituted benzimidazoles.Biophys. J. 16:143
Laprade, R., Ciani, S.M., Eisenman, G., Szabo, G. 1974. The kinetics of carrier-mediated ion permeation in lipid bilayers and its theoretical interpretation.In: Membranes — A Series of Advances. G. Eisenman, Editor. Vol. 3. M. Dekker, New York (In press)
Läuger, P., Stark, G. 1970. Kinetics of carrier-mediated ion transport across lipid bilayer membranes.Biochim. Biophys. Acta 211:458
Lev, A.A., Buzhinsky, E.P. 1967. Cation specificity of the model bimolecular phospholipid membranes with incorporated valinomycin.Tsitologiya 9:102
Liberman, E.A., Topaly, V.P. 1968. Selective transport of ions through bimolecular phospholipid membranes.Biochim. Biophys. Acta 163:125
McLaughlin, S. 1972. The mechanism of action of DNP on phospholipid bilayer membranes.J. Membrane Biol. 9:361
Mueller, P., Rudin, D.O. 1967. Development of K+−Na+ discrimination in experimental bimolecular lipid membranes by macrocyclic antibiotics.Biochim. Biophys. Res. Commun. 26:398
Pinkerton, M., Steinrauf, L.K., Dawkins, P. 1969. The molecular structure and some transport properties of valinomycin.Biochem. Biophys. Res. Commun. 35:512.
Shemyakin, M.M., Ovchinnikov, Yu.A., Ivanov, V.T., Antonov, V.K., Vinogradova, E.I., Shkrob, A.M., Malenkov, G.G., Evstratov, A.V., Laine, I.A., Melnik, E.I., Ryabova, I.D. 1969. Cyclodepsipeptides as chemical tools for studying ionic transport through membranes.J. Membrane Biol. 1:402
Stark, G., Benz, R. 1971. The transport of potassium through lipid bilayer membranes by the neutral carriers valinomycin and monactin.J. Membrane Biol. 5:133
Stark, G., Ketterer, B., Benz, R., Läuger, P.. 1971. The rate constants of valinomycin-medicated ion transport through thin lipid membranes.Biophys. J. 11:981
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kuo, K.H., Bruner, L.J. Modification of valinomycin-mediated bilayer membrane conductance by 4,5,6,7-tetrachloro-2-methylbenzimidazole. J. Membrain Biol. 26, 385–403 (1976). https://doi.org/10.1007/BF01868885
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01868885