Skip to main content
Log in

Acetylcholine receptor-controlled ion flux in electroplax membrane vesicles: Identification and characterization of membrane properties that affect ion flux measurements

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

Summary

Several intrinsic properties of acetylcholine receptor-rich membrane vesicles prepared fromElectrophorus electricus, which need to be considered in measurements of receptor-mediated ion flux, have been identified. One of these properties is a slow exchange of inorganic ions in the vesicles. The slow exchange of ions is not related to the receptor-mediated flux of ions and accounts for 30–35% of the efflux observed. A method to separate this process from the receptor-controlled flux has been developed. It has also been shown, using a light-scattering method, that aggregation-disaggregation of the vesicles can interfere with the efflux measurements, and a method to overcome this problem has been developed. The difference in the amplitude of effluxes induced by saturating amounts of carbamylcholine and gramicidin has been investigated and has been shown not to be due to a receptor-controlled process; therefore, the amplitude difference does not need to be considered in understanding the receptor-controlled process.

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

Access this article

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

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Aoshima, H., Cash, D.J., Hess, G.P. 1980. Acetylcholine receptor-controlled ion flux in electroplax membrane vesicles: A minimal mechanism based on rate measurements in the millisecond to minute time region.Biochem. Biophys. Res. Commun. 92:896

    Article  PubMed  Google Scholar 

  • Bernhardt, J., Neumann, E. 1978. Kinetic analysis of receptor-controlled tracer efflux from sealed membrane fragments.Proc. Nat. Acad. Sci. USA 75:3756

    Google Scholar 

  • Cash, D.J., Hess, G.P. 1980. On the molecular mechanism of acetylcholine receptor-controlled ion translocation across cell membranes.Proc. Nat. Acad. Sci. USA 77:842

    PubMed  Google Scholar 

  • Cash, D.J., Hess, G.P. 1981. Quenched flow technique with plasma membrane vesicles: Acetylcholine receptor mediated transmembrane ion flux.Anal. Biochem. (in press)

  • Cass, A., Finkelstein, A., Krespi, Y. 1970. The ion permeability induced in thin lipid membranes by the polyene antibiotics nystatin and amphotericin B.J. Gen. Physiol. 56:100

    Article  PubMed  Google Scholar 

  • Doty, P., Edsall, J.T. 1951. Light scattering in protein solution.Adv. Protein Chem. 6:35

    PubMed  Google Scholar 

  • Epstein, N., Hess, G.P., Kim, P.S., Noble, R.L. 1980. Inactivation (desensitization) of the acetylcholine receptor inElectrophorus electricus membrane vesicles by carbamylcholine: Comparison between ion flux and α-bungarotoxin binding.J. Membrane Biol. 56:133

    Google Scholar 

  • Fu, J.-j.L., Donner, D.B., Moore, D.E., Hess, G.P. 1977. Allosteric interactions between the membrane-bound acetylcholine receptor and chemical mediators: Equilibrium measurements.Biochemistry 16:678

    Article  PubMed  Google Scholar 

  • Gent, M.P.N., Prestegard, J.H. 1976. Interaction of the polyene antibiotics with lipid bilayer vesicles containing cholesterol.Biochim. Biophys. Acta 426:17

    PubMed  Google Scholar 

  • Hanley, M.R. 1978. Crotoxin effects inTorpedo californica, cholinergic excitable vesicles and the role of its phospholipase activity.Biochem. Biophys. Res. Commun. 82:392

    Article  PubMed  Google Scholar 

  • Hess, G.P. 1979. Acetylcholine receptor-controlled ion fluxes in microsacs (membrane vesicles) obtained from the electroplax ofElectrophorus electricus.In: The Neurosciences: Fourth Study Program. F.O. Schmitt and F.G. Worden, editors, Chapter 49, p. 847. MIT Press, Cambridge

    Google Scholar 

  • Hess, G.P., Andrews, J.P. 1977. Functional acetylcholine receptor-electroplax membrane microsacs (vesicles): Purification and characterization.Proc. Nat. Acad. Sci. USA 74:482

    PubMed  Google Scholar 

  • Hess, G.P., Andrews, J.P., Struve, G.E. 1976) Apparent cooperative effects in acetylcholine receptor-mediated ion flux in electroplax membrane preparations.Biochem. Biophys. Res. Commun. 69:830

    Article  PubMed  Google Scholar 

  • Hess, G.P., Andrews, J.P., Struve, G.E., Coombs, S.E. 1975. Acetylcholine receptor-mediated ion flux in electroplax membrane preparations.Proc. Nat. Acad. Sci. USA 72:4371

    PubMed  Google Scholar 

  • Hess, G.P., Cash, D.J., Aoshima, H. 1979. Acetylcholine receptor-controlled ion fluxes in membrane vesicles investigated by fast reaction techniques.Nature (London) 282:329

    Google Scholar 

  • Hess, G.P., Lipkowitz, S., Struve, G.E. 1978. Acetylcholine-receptor-mediated ion flux in electroplax membrane microsacs (vesicles): Change in mechanism produced by asymmetrical distribution of sodium and potassium ions.Proc. Nat. Acad. Sci. USA 75:1703

    PubMed  Google Scholar 

  • Hsiao-Ping, H., Moore, H., Hartig, P.R., Raftery, M.A. 1979. Correlation of polypeptide composition with functional events in acetylcholine receptor-enriched membranes fromTorpedo californica.Proc. Nat. Acad. Sci. USA 76:6265

    PubMed  Google Scholar 

  • Kasai, M., Changeux, J.-P. 1971a.In vitro excitation of purified membrane fragments by cholinergic agonists. I. Pharmacological properties of the excitable membrane fragments.J. Membrane Biol. 6:1

    Google Scholar 

  • Kasai, M., Changeux, J.-P. 1971b.In vitro excitation of purified membrane fragments by cholinergic agonists. II. The permeability change caused by cholinergic agonists.J. Membrane Biol. 6:24

    Google Scholar 

  • Kasai, M., Changeux, J.-P., 1971c.In vitro excitation of purified membrane fragments by cholinergic agonists. III. Comparison of the dose-response curves to decamethonium with the corresponding binding curves of decamethonium to the cholinergic receptor.J. Membrane Biol. 6:58

    Google Scholar 

  • Katz, B., Thesleff, S. 1957. A study of the “desensitization” produced by acetylcholine at the motor endplate.J. Physiol. (London) 138:64

    Google Scholar 

  • Keynes, R.D., Martins-Ferreira, H. 1953. Membrane potentials in the electroplates of the electric eel.J. Physiol. (London) 119:315

    Google Scholar 

  • Kim, P.S. 1979. Acetylcholine receptor rich electroplax membrane microsacs (vesicles): Identification and characterization of membrane properties that interfere with measurements of receptor-mediated ion flux. A.B. Honors Thesis. Cornell University, Ithaca

    Google Scholar 

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

    PubMed  Google Scholar 

  • Palfrey, C., Littauer, V.Z. 1976. Sodium-dependent efflux of K+ and Rb+ through the activated sodium channel of neuroblastoma cells.Biochem. Biophys. Res. Commun. 72:209

    PubMed  Google Scholar 

  • Popot, J.L., Sugiyama, H., Changeux, J.-P. 1976. Studies on the electrogenic action of acetylcholine withTorpedo marmorata electric organ. II. The permeability response of the receptor-rich membrane fragments to cholinergic agonistsin vitro.J. Mol. Biol. 106:469

    PubMed  Google Scholar 

  • Pressman, B. 1976. Biological applications of ionophores.Annu. Rev. Biochem. 45:501

    PubMed  Google Scholar 

  • Schiebler, W., Hucho, F. 1978. Membranes rich in acetylcholine receptor: Characterization and reconstitution to excitable membranes from exogenous lipids.Eur. J. Biochem. 85:55

    PubMed  Google Scholar 

  • Selwyn, M.J., Dawson, A.P., Stockdale, M., Gains, N. 1970. Chloride-hydroxide exchange across mitochondrial erythrocyte and artifiecal lipid membranes mediated by trialkyl- andtriphenyltin compounds.Eur. J. Biochem. 14:120

    PubMed  Google Scholar 

  • Sillen, L.G., Martell, A.E. (Compilers) 1964. Stability Constants of Metal Ion Complexes. Special Publication. No.17. The Chemical Society, London

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, P.S., Hess, G.P. Acetylcholine receptor-controlled ion flux in electroplax membrane vesicles: Identification and characterization of membrane properties that affect ion flux measurements. J. Membrain Biol. 58, 203–211 (1981). https://doi.org/10.1007/BF01870906

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01870906

Keywords

Navigation