Skip to main content
SpringerLink
Log in

Polypeptide neurotoxins modify gating and apparent single-channel conductance of veratridine-activated sodium channels in planar lipid bilayers

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

Summary

The effects of scorpion and sea anemone polypeptide toxins on partially purified veratridine (VER)-activated Na channels from rat brain were studied at the single-channel level in planar lipid bilayers. The probability of the VER-activated channel being open (P o ) increased with depolarization;P o was 0.5 at −40 to −50 mV. Saxitoxin (STX) blocked VER-activated channels with an apparent dissociation constant of about 1nm at −45 mV. The apparent single-channel conductance was approximately 9 pS, similar to that seen in VER-activated Na channels from skeletal muscle transverse tubules. Addition of sea anemone or scorpion polypeptide toxins to VER-activated Na channels resulted in a 19% increase in apparent single-channel conductance and a hyperpolarizing shift in theP o vs. V m relation such that the channels were more likely to be open at potentials <40 mV. These effects of the polypeptide toxins on the single-channel properties of VER-activated Na channels may account for the previously described potentiation of VER action by polypeptide toxins.

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

  • Agnew, W.S., Levinson, S.R., Brabson, J.S., Raftery, M.A. 1978. Purification of the tetrodotoxin-binding component associated with the voltage-sensitive Na channel fromElectrophorus electricus electroplax membranes.Proc. Natl. Acad. Sci. USA 75:2606–2610

    PubMed  Google Scholar 

  • Barnes, S., Hille, B. 1988. Veratridine modifies open Na channels.J. Gen. Physiol. 91:421–443

    Article  PubMed  Google Scholar 

  • Bradford, M.M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principles of protein dye binding.Anal. Biochem. 72:248–254

    PubMed  Google Scholar 

  • Catterall, W.A. 1975. Activation of the action potential Na+ ionophore of cultured neuroblastoma cells by veratridine and batrachotoxin.J. Biol. Chem. 250:4053–4059

    PubMed  Google Scholar 

  • Catterall, W.A. 1976. Purification of a toxic protein from scorpion venom which activates the action potential Na+ ionophore.J. Biol. Chem. 251:5528–5536

    PubMed  Google Scholar 

  • Catterall, W.A. 1977a. Activation of the action potential Na+ ionophore by neurotoxins. An allosteric model.J. Biol. Chem. 252:8669–8676

    PubMed  Google Scholar 

  • Catterall, W.A. 1977b. Membrane potential-dependent bidning of scorpion toxin to the action potential Na+ ionophore.J. Biol. Chem. 252:8660–8668

    PubMed  Google Scholar 

  • Catterall, W.A. 1988. Structure and function of voltage-sensitive ion channels.Science 233:653–661

    Google Scholar 

  • Catterall, W.A., Beress, L. 1978. Sea anemone toxin and scorpion toxin share a common receptor site associated with the action potential sodium ionophore.J. Biol. Chem. 253:7393–7396

    PubMed  Google Scholar 

  • Corbett, A.M., Zinkand, W.C., Krueger, B.K. 1987. Activation of single neuronal sodium channels by veratridine and polypeptide neurotoxin in planar lipid bilayersBiophys. J. 51:435a (Abstr.)

    Google Scholar 

  • Cukierman, S., Zinkand, W.C., French, R.J., Krueger, B.K. 1988. Effects of membrane surface charge and calcium on the gating of rat brain sodium channels in planar bilayers.J. Gen. Physiol. 92:431–447

    Article  PubMed  Google Scholar 

  • French, R.J., Worley, J.F., Blaustein, M.B., Romine, W.O., Tam, K.K., Krueger, B.K. 1986. Gating of batrachotoxin activated sodium channels in lipid bilayers.In: Ion Channel Reconstitution. C. Miller, editor. pp. 363–383. Plenum, New York

    Google Scholar 

  • Garber, S.S., Miller, C. 1987. Single Na+ channels activated by veratridine and batrachotoxin.J. Gen. Physiol. 89:459–480

    Article  PubMed  Google Scholar 

  • Hanke, W. 1986. Incorporation of ion channels by fusion.In: Ion Channel Reconstitution. C. Miller, editor. pp. 141–153. Plenum, New York

    Google Scholar 

  • Hartshorne, R.P., Catterall, W.A. 1984. The sodium channel from rat brain. Purification and subunit composition.J. Biol. Chem. 259:1667–1675

    PubMed  Google Scholar 

  • Hartshorne, R.P., Keller, B.U., Talvenheimo, J.A., Catterall, W.A., Montal, M. 1985. Functional reconstitution of the purified brain sodium channel in planar bilayers.Proc. Natl. Acad. Sci. USA 82:240–244

    PubMed  Google Scholar 

  • Jacques, Y., Fosset, M., Lazdunski, M. 1978. Molecular properties of the action potential Na+ ionophore in neuroblastoma cells.J. Biol. Chem. 253:7383–7392

    PubMed  Google Scholar 

  • Kopeyan, C., Martinez, G., Rochat, H. 1978. Amino acid sequence of neurotoxin V from the scorpionLeiurus quinquestriatus quinquestriatus.FEBS Lett. 89:54–58

    Article  PubMed  Google Scholar 

  • Krueger, B.K., Blaustein, M.P. 1980. Sodium channels in presynaptic nerve terminals: Regulation by neurotoxins.J. Gen. Physiol. 76:287–313

    Article  PubMed  Google Scholar 

  • Krueger, B.K., Ratzlaff, R.W., Strichartz, G.R., Blaustein, M.P. 1979. Saxitoxin binding to synaptosomes, membranes, and solubilized binding sites from rat brain.J. Membrane Biol. 50:287–310

    Google Scholar 

  • Krueger, B.K., Worley, J.F., French, R.J. 1983. Single sodium channels from rat brain incorporated into planar lipid bilayer membranes.Nature (London) 303:172–175

    Article  Google Scholar 

  • Leibowitz, M.D., Sutro, J.B., Hille, B. 1986. Voltage dependent gating of veratridine-modified Na channels.J. Gen. Physiol. 87:25–46

    Article  PubMed  Google Scholar 

  • Meves, H., Simard, J.M., Watt, D.D. 1986. Interactions of scorpion toxins with the sodium channel.Ann. NY Acad. Sci. 479:113–132

    PubMed  Google Scholar 

  • Miller, C. 1978. Voltage-gated cation conductance channel from fragmented sarcoplasmic reticulum: Steady-state electrical properties.J. Membrane Biol. 40:1–23

    Google Scholar 

  • Moczydlowski, E., Garber, S.S., Miller, C. 1984. Batrachotoxin-activated Na+ channels in planar lipid bilayers. Competition of tetrodotoxin block by Na+.J. Gen. Physiol. 84:665–686

    Article  PubMed  Google Scholar 

  • Mueller, P., Rudin, D.O., Tien, H.T., Wescott, W.C. 1963. Methods for the formation of single bimolecular lipid membranes in aqueous solution.J. Phys. Chem. 67:534–535

    Google Scholar 

  • Nagy, K. 1988. Mechanism of inactivation of single sodium channels after modification by chloramine-T, sea anemone toxin and scorpion toxin.J. Membrane Biol. 106:29–40

    Google Scholar 

  • Norton, T.R., Shibata, S., Kashiwagi, M., Bentley, J. 1976. Isolation and characterization of the cardiotonic polypeptide anthopleurin-A from the sea anemoneAnthopleura xanthogrammica.J. Pharm. Sci. 65:1368–1374

    PubMed  Google Scholar 

  • Patlak, J., Horn, R. 1982. Effect of N-bromoacetamide on single sodium channel currents in excised membrane patches.J. Gen. Physiol. 79:333–351

    PubMed  Google Scholar 

  • Rack, M., Richter, D., Rubly, N. 1987. Purification and characterization of a β-toxin from the venom of the African scorpionLeiurus quinquestriatus.FEBS Lett. 214:163–166

    PubMed  Google Scholar 

  • Romey, G., Renaud, J.F., Fosset, M., Lazdunski, M. 1980. Pharmacological properties of the interaction of a sea anemone polypeptide toxin with cardiac cells in culture.J. Pharmacol. Exp. Ther. 213:607–615

    PubMed  Google Scholar 

  • Rosemblatt, M., Hidalgo, C., Vergara, C., Ikemoto, N. 1981. Immunological and biochemical properties of transverse tubule membranes isolated from rabbit skeletal muscle.J. Biol. Chem. 256:8140–8148

    PubMed  Google Scholar 

  • Sigel, E. 1987. Effects of veratridine on single neuronal sodium channels expressed inXenopus oocytes.Pfluegers Arch. 410:112–120

    Google Scholar 

  • Strichartz, G., Rando, T., Wang, G.K. 1987. An integrated view of the molecular toxinology of sodium channel gating in excitable cells.Ann. Rev. Neurosci. 10:237–267

    PubMed  Google Scholar 

  • Sutro, J.B. 1986. Kinetics of veratridine action on Na channels of skeletal muscle.J. Gen. Physiol. 87:1–24

    PubMed  Google Scholar 

  • Tamkun, M.M., Catterall, W.A. 1981. Ion flux studies of voltage-sensitive sodium channels in synaptic nerve ending particles.Molec. Pharmacol. 19:78–86

    Google Scholar 

  • Tanaka, M., Haniu, M., Yasanobu, K.T., Norton, T.R. 1977. Amino acid sequence of theAnthopleura xanthogrammica heart stimulant, Anthopleurin A.Biochemistry 16:204–208

    PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Physiology, University of Maryland School of Medicine, 21201, Baltimore, Maryland

    Adrian M. Corbett & Bruce K. Krueger

Authors
  1. Adrian M. Corbett
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bruce K. Krueger
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Corbett, A.M., Krueger, B.K. Polypeptide neurotoxins modify gating and apparent single-channel conductance of veratridine-activated sodium channels in planar lipid bilayers. J. Membrain Biol. 110, 199–207 (1989). https://doi.org/10.1007/BF01869150

Download citation

  • Received:

  • Revised:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01869150

Key words

Search

Navigation