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Intracellular binding of cationized ferritin prolongs the time course of sodium channel inactivation in squid giant axons

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Summary

Cationized ferritin (CF) applied intracellularly in squid giant axons bound to the negatively charged sites on the cytoplasmic surface of the axolemma. Under the electron microscope, the distribution of CF was found to be dense and uniform over the axolemmal surface. However, the effect of CF on the membrane excitability was highly specific, the major effect being a prolonging of the inactivation time course of the sodium channel without altering the properties of the potassium channel. The binding of CF did not alter the surface potential related to the membrane excitability. When CF was present intracellularly, the time course of the inactivation was characterized by two time constants (slow and fast). The slow component increased with an increase in CF binding and its time constant had a unique value (26 msec) irrespective of the duration of perfusion and concentration of CF. The concentration of CF at which the half-maximum response occurred was about 150nm. Poly-l-glutamate, charged negatively at neutral pH, removed CF from the axolemma and counteracted the CF effect on the sodium channel, although this poly-acidper se did not affect the membrane excitability. Our results indicate that CF binds electrostatically to the inactivation site of the sodium channel but does not affect the voltage sensor, which is supposed to be located deep in the channel.

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References

  1. Armstrong, C.M., Bezanilla, F., Rojas, E. 1973. Destruction of sodium conductance inactivation in squid axons perfused with pronase.J. Gen. Physiol. 62:375–391

    Google Scholar 

  2. Baker, P.F., Hodgkin, A.L., Shaw, T.I. 1962. Replacement of the axoplasm of giant nerve fibres with artificial solutions.J. Physiol. (London) 164:330–354

    Google Scholar 

  3. Carbone, E., Fioravanti, R., Prestipino, G., Wanke, E. 1978. Action of extracellular pH on Na+ and K+ membrane currents in the giant axon ofLoligo vulgaris.J. Membrane Biol. 43:295–315

    Google Scholar 

  4. Carbone, E., Testa, P.L., Wanke, E. 1981. Intercellular pH and ionic channels in theLoligo vulgaris giant axon.Biophys. J. 35:393–413

    Google Scholar 

  5. Chandler, W.K., Hodgkin, A.L., Meves, H. 1965. The effect of changing the internal solution on sodium inactivation and related phenomena in giant axons.J. Physiol. (London) 180:821–836

    Google Scholar 

  6. Clegg, G.A., Fitton, J.E., Harrison, P.M., Treffry, A. 1980. Ferritin: Molecular structure and iron-storage mechanisms.Prog. Biophys. Mol. Biol. 36:56–86

    Google Scholar 

  7. Danon, D., Goldstein, L., Marikovsky, Y., Skutelsky, E. 1972. Use of cationized ferritin as a label of negative charges on cell surfaces.J. Ultrastruct. Res. 38:500–510

    Google Scholar 

  8. D'Arrigo, J.S. 1973. Possible screening of surface charges on crayfish axons by polyvalent metal ions.J. Physiol. (London) 231:117–128

    Google Scholar 

  9. Drouin, H., Neumcke, B. 1974. Specific and unspecific charges at the sodium channels of the nerve membrane.Pfluegers Arch. 351:207–229

    Google Scholar 

  10. Fohlmeister, J.F., Adelman, W.J., Jr. 1982. Periaxonal surface calcium binding and distribution of charge on the faces of squid axon potassium channel molecules.J. Membrane Biol. 70:115–123

    Google Scholar 

  11. Frace, A.M., Poznansky, M., Eaton, D.C., Brodwick, M.S. 1985. Effects of phospholipid headgroup modification in voltage clamped squid axon.Biophys. J. 47:440a

    Google Scholar 

  12. Frankenhaeuser, B., Hodgkin, A.L. 1957. The action of calcium on the electrical properties of squid axons.J. Physiol. (London) 137:218–244

    Google Scholar 

  13. Furuya, K., Furuya, S., Yamagishi, S. 1983. Developmental time courses of Na and Ca spikes in neuroblastoma x glioma hybrid cells.Dev. Brain Res. 11:229–234

    Google Scholar 

  14. Gilbert, D.L., Ehrenstein, G. 1969. Effect of divalent cations on potassium conductance of squid axons: Determination of surface charge.Biophys. J. 9:447–463

    Google Scholar 

  15. Hahin, R., Campbell, D.T. 1983. Simple shifts in the voltage dependence of sodium channel gating caused by divalent cations.J. Gen Physiol. 82:785–805

    Google Scholar 

  16. Hille, B. 1968. Charges and potentials at the nerve surface. Divalent ions and pH.J. Gen. Physiol. 51:221–236

    Google Scholar 

  17. Hille, B., Woodhull, A. M., Shapiro, B.I. 1975. Negative surface charge near sodium channels of nerve: Divalent ions, monovalent ion, and pH.Philos. Trans. R. Soc. London B 270:301–318

    Google Scholar 

  18. Hirano, H., Nishiyama, F., Furuya, K., Yamagishi, S. 1983. Binding of cationized ferritin on the cytoplasmic surface of the axolemma and its effect on the membrane excitation.Proc. Jpn. Acad. B59:140–143

    Google Scholar 

  19. Kukita, F. 1982. Properties of sodium and potassium channels of the squid giant axon far below 0°C.J. Membrane Biol. 68:151–160

    Google Scholar 

  20. Moore, J.W., Narahashi, T., Ulbricht, W. 1964. Sodium conductance shift in an axon internally perfused with a sucrose and low-potassium solution.J. Physiol. (London) 172:163–172

    Google Scholar 

  21. Mozhayeva, G.N., Naumov, A.P. 1970. Effect of surface charge on the steady-state potassium conductance of nodal membrane.Nature (London) 228:164–165

    Google Scholar 

  22. Okamoto, H., Takahashi, K., Yamashita, N. 1977. One-to-one binding of a purified scorption toxin to Na channels.Nature (London) 266:465–468

    Google Scholar 

  23. Oxford, G.S., Wu, C.H., Narahashi, T. 1978. Removal of sodium channel inactivation in squid giant axons byN-bromoacetamide.J. Gen. Physiol. 71:227–247

    Google Scholar 

  24. Rennke, H.G., Cotran, R.S., Venkatachalam, M.A. 1975. Role of molecular charge in glomerular permeability. Tracer studies with cationized ferritins.J. Cell Biol. 67:638–646

    Google Scholar 

  25. Takata, K., Nishiyama, F., Hirano, H. 1981. Double labeling study of anionic sites and concanavalin A binding sites in monkey macrophages.J. Histochem. Cytochem. 29:858–863

    Google Scholar 

  26. Takenaka, T., Yamagishi, S. 1969. Morphology and electrophysiological properties of squid giant axons perfused intracellularly with protease solution.J. Gen. Physiol. 53:81–96

    Google Scholar 

  27. Tasaki, I., Watanabe, A., Takenaka, T. 1962. Resting and action potential of intracellularly perfused squid giant axon.Proc. Natl. Acad. Sci. USA 48:1177–1184

    Google Scholar 

  28. Wang, G.K. 1984. Irreversible modification of sodium channel inactivation in toad myelinated nerve fibres by the oxidant chloramine-T.J. Physiol. (London) 346:127–141

    Google Scholar 

  29. Wanke, E., Carbone, E., Testa, P.L. 1980. The sodium channel and intracellular H+ blockage in squid axons.Nature (London) 287:62–63

    Google Scholar 

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Furuya, K., Hirano, H., Nishiyama, F. et al. Intracellular binding of cationized ferritin prolongs the time course of sodium channel inactivation in squid giant axons. J. Membrain Biol. 89, 75–83 (1986). https://doi.org/10.1007/BF01870897

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  • DOI: https://doi.org/10.1007/BF01870897

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