Ionophore-mediated transmembrane movement of divalent cations in small unilamellar liposomes: An evaluation of the chlortetracycline fluorescence technique and correlations with black lipid membrane studies
- 52 Downloads
- 7 Citations
Summary
Conceptual advances in the field of membrane transport have, in the main, utilized artificial membranes, both planar and vesicular. Systems of biological interest,viz., cells and organelles, resemble vesicles in size and geometry. Methods are, therefore, required to extend the results obtained with planar membranes to liposome systems. In this report we present an analysis of a fluorescence technique, using the divalent cation probe chlortetracycline, in small, unilamellar vesicles, for the study of divalent cation fluxes. An ion carrier (X537 A) and a pore former (alamethicin) have been studied. The rate of rise of fluorescence signal and the transmembrane ion gradient have been related to transmembrane current and potential, respectively. A second power dependence of ion conduction-including the electrically silent portion thereof — on X537 A concentration, has been observed. An exponential dependence of “current” on “transmembrane potential” in the case of alamethicin is also confirmed. Possible errors in the technique are discussed.
Key words
Membrane-transport fluorescence chlortetracycline X537 A alamethicin ionophore liposomePreview
Unable to display preview. Download preview PDF.
References
- Bangham, A.D., Hill, M.W., Miller, N.G.A. 1974. Preparation and use of liposomes as models of biological membranes.In: Methods in Membrane Biology. E.D. Korn, editor. Vol. 1, pp. 1–68, Plenum, New YorkGoogle Scholar
- Bangham, A.D., Standish, N.M., Watkins, J.C. 1965. Diffusion of univalent ions across the lamellae of swollen phospholipids.J. Mol. Biol. 13:238–252Google Scholar
- Bean, R.C., Shephered, W.C., Chan, H., Eichner, J. 1969. Discrete conductance fluctuations in lipid bilayer protein membranes.J. Gen. Physiol. 53:741–757Google Scholar
- Boheim, G., Kolb, H.-A. 1978. Analysis of multi-pore systems of alamethicin in a lipid membrane. I. Voltage-jump current-relaxation measurements.J. Membrane Biol. 38:99–150Google Scholar
- Brunner, J., Skrabal, P., Hauser, H. 1976. Single bilayer vesicles prepared without sonication. Physico-chemical properties.Biochim. Biophys. Acta 455:322–331Google Scholar
- Caswell, A.H., Hutchison, J.D. 1971. Selectivity of cation chelation to tetracyclines: Evidence for special conformation of calcium chelate.Biochem. Biophys. Res. Commun. 43:625–630Google Scholar
- Caswell, A.H., Pressman, B.C. 1972. Kinetics of transport across sarcoplasmic reticulum vesicles induced by ionophores.Biochem. Biophys. Res. Commun. 49:292–298Google Scholar
- Caswell, A.H., Warren, S. 1972. Observation of Ca2+ uptake by isolated sarcoplasmic reticulum employing a fluorescent chelate probe.Biochem. Biophys. Res. Commun. 46:1757–1763Google Scholar
- Célis, H., Estrada-O., S., Montal, M. 1974. Model translocators for divalent and monovalent ion transport in phospholipid membranes. I. The ion permeability induces in lipid bilayers by the antibiotic X-537 A.J. Membrane Biol. 18:187–199Google Scholar
- Eisenberg, M., Hall, J.E., Mead, C.A. 1973. The nature of the voltage-dependent conductance induced by alamethicin in black lipid membranes.J. Membrane Biol. 14:143–176Google Scholar
- Fringeli, U.P. 1980. Distribution and diffusion of alamethicin in a lecithin/water model membrane system.J. Membrane Biol. 54:203–212Google Scholar
- Fringeli, U.P., Fringeli, M. 1979. Pore formation in lipid membranes by alamethicin.Proc. Natl. Acad. Sci. USA 76:3852–3856Google Scholar
- Hladky, S.B., Rink, T.F. 1976. Membrane potentials and properties of human erythrocytes and ghosts assessed with a fluorescent dye, 3,3′-dipropyl-2,2′ thiadicarbocyanine.J. Physiol. (London) 258:100PGoogle Scholar
- Johnson, S.M., Bangham, A.D. 1969. Potassium permeability of single compartment liposomes with and without valinomycin.Biochim. Biophys. Acta 193:82–91Google Scholar
- Johnson, S.M., Herrin, J., Lin, S.J., Paul, I.C. 1970. The crystal and molecular structure of the barium salt of an antibiotic containing a high proportion of oxygen.J. Am. Chem. Soc. 92:4428–4435Google Scholar
- Mathew, M.K., Balaram, P. 1980. A reinvestigation of chlortetracycline fluorescence: Effect of pH, metal ions and environment.J. Inorg. Biochem. 13:339–346Google Scholar
- Mueller, P. 1976. Molecular aspects of electrical excitation in lipid bilayers and cell membranes.In: Horizons in Biochemistry and Biophysics. E. Quagliariello, F. Palmieri, and T.P. Singer, editors. Vol. 2, pp. 230–284. Addison-Wesley, Reading (Mass.)Google Scholar
- Mueller, P., Rudin, D.O. 1967. Development of K+−Na+ discrimination in experimental bimolecular lipid membranes by macrocyclic antibiotics.Biochem. Biophys. Res. Commun. 26:398–404Google Scholar
- Mueller, P., Rudin, D.O. 1968. Action potentials induced in bimolecular lipid membranes.Nature (London) 217:713–719Google Scholar
- Mueller, P., Rudin, D.O., Tien, H.Ti., Wescott, W.C. 1962. Reconstitution of cell membrane structurein vitro and its transformation into an excitable system.Nature (London) 294:979–980Google Scholar
- Nagaraj, R., Balaram, P. 1981. Solution phase synthesis of alamethicin I.Tetrahedron 37:1263–1270Google Scholar
- Nagaraj, R., Mathew, M.K., Balaram, P. 1980. Cation translocating effects of alamethicin and its synthetic fragments in lipid membranes. Influence of peptide chain length and charge.FEBS Lett. 121:365–368Google Scholar
- Pressman, B.C. 1973. Properties of ionophores with broad range cation selectivity.Fed. Proc. 32:1698–1703Google Scholar
- Schadt, M., Haeusler, G. 1974. Permeability of lipid bilayer membranes to biogenic amines and cations: Changes induced by ionophores and correlations with biological activities.J. Membrane Biol. 18:277–294Google Scholar
- Sehlin, J., Taeljedahl, I.B. 1979.45Ca, ion uptake by dispersed pancreatic islet cells: Effects ofd-glucose and chlortetracycline.Pfluegers Arch. 381:281–285Google Scholar
- Sten-Knudsen, O. 1978. Passive transport processes.In: Membrane Transport in Biology. G. Giebisch, D.C. Tosteson, and H.H. Ussing, editors. Vol. 1, pp. 5–113. Springer-Verlag, BerlinGoogle Scholar
- Szoka, F., Papahadjopoulos, D. 1980. Comparative properties and methods of preparation of lipid vesicles (liposomes).Annu. Rev. Biophys. Bioeng. 9:467–508Google Scholar