Skip to main content
SpringerLink
Log in

The effect of ethanol on lateral and rotational mobility of plasma membrane vesicles isolated from cultured mar 18.5 hybridoma cells

  • Articles
  • Published:
The Journal of Membrane Biology Aims and scope Submit manuscript

Abstract

Intramolecular excimer formation of 1,3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to evaluate the effect of ethanol on the rate and range of the lateral mobility and the range of the rotational mobility of bulk bilayer structures of the plasma membrane vesicles (ATCC-PMV) isolated from cultured hybridoma cells (ATCC TIB 216). In a concentration-dependent manner, ethanol increased the excimer to monomer fluorescence intensity ratio (I′/I) of Py-3-Py in the ATCC-PMV and decreased the anisotropy (r), limiting anisotropy (r∞) and order parameter (S) of DPH in the ATCC-PMV. This indicates that ethanol increased both the lateral and rotational mobility of the probes in the ATCC-PMV. Selective quenching of DPH by trinitrophenyl groups was utilized to examine the range of transbilayer asymmetric rotational diffusion of the ATCC-PMV. The anisotropy (r), limiting anisotropy (r ∞) and order parameter (S) of DPH in the inner monolayer were 0.024, 0.032, and 0.069, respectively, greater than calculated for the outer monolayer of the ATCC-PMV. Selective quenching of DPH by trinitrophenyl groups was also used to examine the transbilayer asymmetric effects of ethanol on the range of the rotational mobility of the ATCC-PMV. Ethanol had a greater increasing effect on the range of the rotational mobility of the outer monolayer as compared to the inner monolayer of the ATCC-PMV. It has been proven that ethanol exhibits a selective rather than nonselective fluidizing effect within the transbilayer domains of the ATCC-PMV.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Bangham, A.D., Mason, W. 1979. The effect of some general anesthetics on the surface potential of lipid monolayers. Br. J. Pharmacol. 66:259–265

    Google Scholar 

  • Chabanel, A., Abbott, R.E., Chien, S., Schachter, D. 1985. Effects of benzyl alcohol on erythrocyte shape, membrane hemileaflet fluidity, and membrane viscoelasticity. Biochim. Biophys. Acta 816:142–152

    Google Scholar 

  • Chin, J.H., Goldstein, D.B. 1981. Membrane-disordering action of ethanol. Variation with membrane cholesterol content and depth of the spin label probe. Mol. Pharmacol. 19:425–431

    Google Scholar 

  • Chung, I.K., Kang, J.S., Yun, I. 1993a. The effect of n-alkanols on the lateral diffusion of synaptosomal plasma membrane vesicles isolated from bovine cerebral cortex. Korean J. Phannacol. 29:157–163

    Google Scholar 

  • Chung, I.K., Kang, J.S., Yun, I. 1993b. Effects of n-alkanols on the lateral diffusion of total lipid and phospholipid fraction extracted from brain membranes. Arch. Pharm. Res. 16:191–195

    Google Scholar 

  • Chung, Y.Z., Cho, G.J., Yun, I. 1993c. Effects of n-alkanols on the rotational relaxation time of 1,6-diphenyl-1,3,5-hexatriene in the synaptosomal plasma membrane vesicles isolated from bovine cerebral cortex. Arch. Pharm. Res. 16:118–122

    Google Scholar 

  • Chung, Y.Z., Son, H.C., Kim, B.S., Kim, J.S., Kim, M.H., Kang, J.S., Yun, I. 1993d. Effects of n-alkanols on the rotational relaxation time of 1,6-diphenyl-1,3,5-hexatriene in the model membranes of phospholipids extracted from brain membranes. Mol. Cells 3:183–188

    Google Scholar 

  • Franks, N.P., Lieb, W.R. 1987. What is the molecular nature of general anaesthetic target sites? TIPS 8:169–174

    Google Scholar 

  • Gonzales, R.A., Hoffman, P.L. 1991. Receptor-gated ion channels may be selective CNS targets for ethanol. TIPS 12:1–3

    Google Scholar 

  • Harris, R.A., Schroeder, F. 1981. Ethanol and physical properties of brain membranes. Fluorescence studies. Mol. Pharmacol. 20:128–137

    Google Scholar 

  • Hitzemann, R.J., Schueler, H.E., Graham-Brittain, C., Kreishman, G.P. 1986. Ethanol-induced changes in neuronal membrane order. An NMR study. Biochim. Biophys. Acta 859:189–197

    Google Scholar 

  • Jain, M.K., Wu, N.M. 1977. Effect of small molecules on the dipalmitoyl liposomal bilayer: III. Phase transition in lipid bilayer. J. Membrane Biol. 34:157–201

    Google Scholar 

  • Janoff, A.S., Boni, L.T., Rauch, J. 1988. Phase-defined lipid domains in biological membranes: a perspective. In: Advances in Membrane Fluidity. R.C. Aloia, C.C. Curtain, L.M. Gordon, editors. Vol. 2, pp. 101–109. Alan R. Liss, New York

    Google Scholar 

  • Kawato, S., Kinosita, K., Ikegami, A. 1978. Effect of cholesterol on the molecular motion in the hydrocarbon region of lecithin bilayers studied by nanosecond fluorescence techniques. Biochemistry 17:5026–5031

    Google Scholar 

  • Lakowicz, J.R., Prendergast, F.G., Hogen, D. 1979. Fluorescence anisotropy measurements under oxygen quenching conditions as a method to quantify the depolarizing rotations of fluorophores. Application to diphenylhexatriene in isotropic solvents and in lipid bilayers. Biochemistry 18:520–527

    Google Scholar 

  • Lee, N.M., Becker, C.E. 1988. The alcohols. In: Basic and Clinical Pharmacology. B.G. Katzung, editor., pp. 278–286. Prentice-Hall of Japan, Tokyo

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.R., Farr, A.L., Randall, R.J. 1951. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193:265–275

    CAS  PubMed  Google Scholar 

  • Majchrowicz, E., Mendelson, J.H. 1971. Blood methanol concentrations during experimentally induced ethanol intoxication in alcoholics. J. Pharmacol. Exp. Ther. 179:293–300

    Google Scholar 

  • Manevich, E.M., Koiv, A., Jarv, J., Molotkovsky, J.G., Bergelson, L.D. 1988. Binding of specific ligands to muscarinic receptors alters the fluidity of membrane fragments from rat brain. A fluorescence polarization study with lipid-specific probes. FEBS Lett. 236:43–46

    Google Scholar 

  • Molitoris, B.A., Hoilien, C. 1987. Static and dynamic components of renal cortical brush border and basolateral membrane fluidity: role of cholesterol. J. Membrane Biol. 99:165–172

    Google Scholar 

  • Sanna, E., Concas, A., Serra, M., Santoro, G., Biggo, G. 1991. Ex vivo binding of t-[35S]butylbicyclophosphorothionate: a biochemical tool to study the pharmacology of ethanol at the γ-aminobutyric acid-coupled chloride channel. J. Pharmacol. Exp. Ther. 256:922–928

    Google Scholar 

  • Schachter, D. 1984. Fluidity and function of hepatocyte plasma membranes. Hepatology 4:140–151

    Google Scholar 

  • Schroeder, F., Morrison, W.J., Gorka, C., Wood, W.G. 1988. Transbilayer effects of ethanol on fluidity of brain membrane leaflets. Biochim. Biophys. Acta 946:85–94

    Google Scholar 

  • Seeman, P. 1972. The membrane actions of anesthetics and tranquilizers. Pharmacol. Rev. 24:585–655

    Google Scholar 

  • Sweet, W.D., Wood, W.G., Schroeder, F. 1987. Charged anesthetics selectively alter plasma membrane order. Biochemistry 26:2828–2835

    Google Scholar 

  • Treistman, S.N., Moynihan, M.M., Wolf, D.E. 1987. Influence of alcohols, temperature, and region on the mobility of lipids in neuronal membrane. Biochim. Biophys. Acta 898:109–120

    Google Scholar 

  • van Blitterwijk, W.J., Van Hoeven, R.P., van der Meer, B.W. 1981. Lipid structural order parameters (reciprocal of fluidity) in biomembranes derived from steady-state fluorescence polarization measurements. Biochim. Biophys. Acta 644:323–332

    Google Scholar 

  • Wood, W.G., Gorka, C., Schroeder, F. 1989. Acute and chronic effects of ethanol on transbilayer membrane domains. J. Neurochem. 52:1925–1930

    Google Scholar 

  • Yun, I., Chung, I.K., Park, Y.M., Kang, J.S. 1993b. Effects of ethanol on the fluidity of plasma membrane vesicles isolated from cultured mouse myeloma cell line Sp2/0-Ag14. Korean J. Pharmacol, 29:149–156

    Google Scholar 

  • Yun, I., Kang, J.S. 1990. The general lipid composition and aminophospholipid asymmetry of synaptosomal plasma membrane vesicles isolated from bovine cerebral cortex. Mol. Cells 1:15–20

    Google Scholar 

  • Yun, I., Kang, J.S. 1992a. Transbilayer effects of n-alkanols on the fluidity of model membranes of total lipids extracted from synaptosomal plasma membrane vesicles. Korean J. Pharmacol. 28:191–199

    Google Scholar 

  • Yun, I., Kang, J.S. 1992b. Transbilayer effects of n-alkanols on the fluidity of phospholipid model membranes. Arch. Pharm. Res. 15:152–161

    Google Scholar 

  • Yun, I., Kim, Y.S., Yu, S.H., Chung, I.K., Kim, I.S., Baik, S.W., Cho, G.J., Chung, Y.Z., Kim, S.H., Kang, J.S. 1990a Comparison of several procedures for the preparation of synaptosomal plasma membrane vesicles. Arch. Pharm. Res. 13:325–329

    Google Scholar 

  • Yun, I., Kim, H.I., Hwang, T.H., Kim, J.R., Kim, I.S., Chung, Y.Z., Shin, Y.H., Jung, H.O., Kang, J.S. 1990b. Effects of barbiturates on the fluidity of phosphatidylethanolamine model membranes. Korean J. Pharmacol. 26:209–217

    Google Scholar 

  • Yun, I., Yang, M.S., Kim, I.S., Kang, J.S. 1993a. Bulk vs. transbilayer effects of ethanol on the fluidity of the plasma membrane vesicles of cultured Chinese hamster ovary cells. Asia Pacific J. Pharmacol. 8:9–15

    Google Scholar 

  • Zachariasse, K.A., Vaz, W.L.C., Sotomayor, C., Kühnle, W. 1982. Investigation of human erythrocyte ghost membranes with intramolecular excimer probes. Biochim. Biophys. Acta 688:323–332

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Dental Pharmacology and Biophysics, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University, 602-739, Pusan, Korea

    I. Yun & S. -H. Lee

  2. Department of Oral Biochemistry and Molecular Biology, College of Dentistry and Research Institute for Oral Biotechnology, Pusan National University, 602-739, Pusan, Korea

    J. -S. Kang

Authors
  1. I. Yun
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. S. -H. Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. J. -S. Kang
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This paper was supported in part by a research grant from the Korea Science and Engineering Foundation (KOSEF 88-1013-01) and from the Korea Research Foundation (1991–1993).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Yun, I., Lee, S.H. & Kang, J.S. The effect of ethanol on lateral and rotational mobility of plasma membrane vesicles isolated from cultured mar 18.5 hybridoma cells. J. Membarin Biol. 138, 221–227 (1994). https://doi.org/10.1007/BF00232794

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00232794

Key words

Search

Navigation