Summary
Valinomycin has been shown to increase the amount of 8-anilino-1-naphtha-lenesulfonate (ANS) bound to egg lecithin liposomes and also to increase the maximum fluorescence value, as derived from double reciprocal plots. The assay conditions were such that addition of valinomycin would not produce a transmembrane potential. The formation of a valinomycin potassium ANS complex in the micelle membrane is proposed. This could account for the increase in the maximum fluorescence value and, by acting as an ANS transporter, could also account for the increase in ANS bound.
Tributylamine was also shown to increase the binding and maximum fluorescence of ANS. In assay conditions where the addition of valinomycin would produce a transmembrane potential negative inside, the tributylamine-induced fluorescence was reversed. The fluorescence decrease is interpreted as transmembrane electrophoresis of ANS in response to a transmembrane potential.
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Ainsworth, S., Flanagan, M.T. 1969. The effect that the environment exerts on the spectroscopic properties of certain dyes that are bound to bovine serum albumin.Biochim. Biophys. Acta 194:213
Azzi, A. 1969. Redistribution of the electrical charge of the mitochondrial membrane during energy conservation.Biochem. Biophys. Res. Commun. 37:254
Azzi, A., Gherardini, P., Santato, M. 1971. Fluorochrome interaction with the mitochondrial membrane. The effect of energy conservation.J. Biol. Chem. 246:2053
Bakker, E.P., Van Dam, K. 1974. The influence of the diffusion potentials across liposomal membranes on the fluorescence intensity of 1-anilinonaphthalene-8-sulphonate.Biochim. Biophys. Acta 339:157
Barker, R.W., Barrett-Bee, K.J., Berden, J.A., McCall, C.E., Radda, G.K. 1974. Sidedness and location of small molecules in membranes.In: Dynamics of Energy Transducing Membranes. L. Ernster, R.W. Estabrook and E.C. Slater, editors. p. 321. Elsevier, Amsterdam
Blok, M.C., De Gier, J., Van Deenen, L.L.M. 1974. Kinetics of the valinomycin-induced potassium ion leak from liposomes with potassium thiocyanate enclosed.Biochim. Biophys. Acta 367:210
Brocklehurst, J.R., Freedman, R.B., Hancock, D.J., Radda, G.K. 1970. Membrane studies with polarity-dependent and excimer-forming fluorescent probes.Biochem. J. 116:721
Chance, B. 1970. Fluorescent probe environment and the structural and charge changes in energy coupling of mitochondrial membranes.Proc. Nat. Acad. Sci. U.S.A. 67:560
Chapman, D., Fluck, D.J., Penkett, S.A., Shipley, G.G. 1968. Physical studies of phospholipids. X. The effect of sonication on aqueous dispersions of egg yolk lecithin.Biochim. Biophys. Acta 163:255
Conti, F., Malerba, F. 1972. Fluorescence signals in ANS-stained lipid bilayers under applied potentials.Biophysik 8:326
Davis, D.G., Tosteson, D.C. 1971. Interaction between valinomycin K+, Na+ and anions in CDCl3 and hexane.Biophys. J. 11:310a
Feinstein, M.B., Felsenfeld, H. 1971. The detection of ionophorous antibiotic-cation complexes in water with fluorescent probes.Proc. Nat. Acad. Sci. U.S.A. 68:2037
Flanagan, M.T., Ainsworth, S. 1968. The binding of aromatic sulphonic acids to bovine serum albumin.Biochim. Biophys. Acta 168:16
Flanagan, M.T., Hesketh, T.R. 1973. Electrostatic interactions in the binding of fluorescent probes to lipid membranes.Biochim. Biophys. Acta 298:535
Fortes, P.A.G., Hoffman, J.F. 1974. The interaction of fluorescent probes with anion permeability pathways of human red cells.J. Membrane Biol. 16:79
Gains, N., Dawson, A.P. 1975. 8-anilino-1-naphthalenesulphonate interaction with whole and disrupted mitochondria. A re-evaluation of the use of double reciprocal plots in the derivation of binding parameters for fluorescent probes binding to mitochondrial membranes.Biochem. J. 148:157
Haynes, D.H. 1972. Detection of ionophore-cation complexes on phospholipid membranes.Biochim. Biophys. Acta 255:406
Jasaitis, A.A., Kuliene, V.V., Skulachev, V.P. 1971. Anilinonaphthalenesulphonate fluorescence changes produced by non-enzymatic generation of membrane potentials in mitochondria and submitochondrial particles.Biochim. Biophys. Acta 234:177
Jasaitis, A.A. La Van Chu, Skulachev, V.P. 1973. Anilino-naphthalene sulphonate and other synthetic ions as mitochondrial membrane penetrants: An H+ pulse technique study.FEBS Lett. 31:241
Johnson, S.M., Bangham, A.D. 1969. Potassium permeability of single compartment liposomes with and without valinomycin.Biochim. Biophys. Acta 193:82
Layton, D.G., Symmons, P., Williams, W.P. 1974. An analysis of the binding of 8-anilino-1-naphthalene-sulphonate to sub-mitochondrial particles.FEBS Lett. 41:1
McLaughlin, S.G.A., Szabo, G., Eisenman, G. 1971. Divalent ions and the surface potential of charged phospholipid membranes.J. Gen. Physiol. 58:667
Mitchell, P. 1970. Discussion.In: Electron Transport and Energy Conservation. J.M. Tager, S. Papa, E. Quagliariello and E.C. Slater, editors. p. 563. Adriatica Editrice, Bari
Penzer, G.R. 1972. The dependence of emission spectra on molecular conformation studied by fluorescence and proton magnetic resonance.Europ. J. Biochem. 25:218
Radda, G.K. 1971. The design and use of fluorescent probes for membrane studies.In: Current Topics in Bioenergetics. D.R. Sanadi, editor. Vol. 4, p. 81. Academic Press, New York
Singleton, W.S., Gray, M.S., Brown, M.L., White, J. 1965. Chromatographically homogenous lecithin from egg phospholipids.J. Amer. Oil Chem. Soc. 42:53
Skulachev, V.P. 1971. Energy transformations in the respiratory chain.In: Current Topics in Bioenergetics. D.R. Sanadi, editor. Vol. 4, p. 127. Academic Press, New York
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Gains, N., Dawson, A.P. Transmembrane electrophoresis of 8-anilino-1-naphthalenesulfonate through egg lecithin liposome membranes. J. Membrain Biol. 24, 237–248 (1975). https://doi.org/10.1007/BF01868625
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DOI: https://doi.org/10.1007/BF01868625