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. 1981 Feb 1;77(2):205–221. doi: 10.1085/jgp.77.2.205

Saxitoxin binding in nerves from walking legs of the lobster Homarus americanus. Two classes of receptors

PMCID: PMC2215429  PMID: 6267162

Abstract

The binding of exchange-labeled saxitoxin (STX) to sodium channels has been investigated in the nonmyelinated fibers of the walking leg nerves of the lobster. The properties of the STX binding site differed systematically among the nerves from different walking legs. The equilibrium dissociation constant for STX binding (KSTX) to the front legs is approximately twice that for the binding to the rear legs; the average ratio of KSTX (front): KSTX (rear) from five separate experiments was 1.80 +/- 0.21 (mean +/- SE). The actual KSTX values ranged from 124.0 to 22.7 nM for the front leg nerves and from 8.6 to 12.7 nM for the rear leg nerves. KSTX values for the middle two walking leg nerves fell between those for the front and rear legs. The inhibitory dissociation constant for tetrodotoxin (KTTX), calculated from tetrodotoxin's inhibition of labeled STX binding, was 3.02 +/- 0.27 nM for the front legs and 2.20 +/- 0.33 nM for the rear legs. The ratio KSTX: KTTX was different in the front and rear leg nerves, being 5.5 and 4.2, respectively. The apparent P pKa of the STX receptor also differed between the two legs, being 4.6 +/- 0.3 for the front legs and 5.1 +/- 0.1 for the rear legs. These results demonstrate that one tissue type in one organism can contain different toxin binding sites. The difference in the receptors can be qualitatively accounted for by the location of an additional negative charge near the receptor site of the rear walking leg.

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Selected References

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  1. Baumgold J. 3H-saxitoxin binding to nerve membranes: inhibition by phospholipase A2 and by unsaturated fatty acids. J Neurochem. 1980 Feb;34(2):327–334. doi: 10.1111/j.1471-4159.1980.tb06601.x. [DOI] [PubMed] [Google Scholar]
  2. Bay C. M., Strichartz G. R. Saxitoxin binding to sodium channels of rat skeletal muscles. J Physiol. 1980 Mar;300:89–103. doi: 10.1113/jphysiol.1980.sp013153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Catterall W. A., Morrow C. S. Binding to saxitoxin to electrically excitable neuroblastoma cells. Proc Natl Acad Sci U S A. 1978 Jan;75(1):218–222. doi: 10.1073/pnas.75.1.218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Colquhoun D., Henderson R., Ritchie J. M. The binding of labelled tetrodotoxin to non-myelinated nerve fibres. J Physiol. 1972 Dec;227(1):95–126. doi: 10.1113/jphysiol.1972.sp010022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Crone H. D., Leake B., Jarvis M. W., Freeman S. E. On the nature of "maculotoxin", a toxin from the blue-ringed octopus (Hapalochlaena maculosa). Toxicon. 1976;14(6):423–426. doi: 10.1016/0041-0101(76)90057-x. [DOI] [PubMed] [Google Scholar]
  6. Drouin H., Neumcke B. Specific and unspecific charges at the sodium channels of the nerve membrane. Pflugers Arch. 1974;351(3):207–229. doi: 10.1007/BF00586919. [DOI] [PubMed] [Google Scholar]
  7. Dulhunty A., Gage P. W. Selective effects of an octopus toxin on action potentials. J Physiol. 1971 Oct;218(2):433–445. doi: 10.1113/jphysiol.1971.sp009626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gage P. W., Moore J. W., Westerfield M. An octopus toxin, maculotoxin, selectively blocks sodium current in squid axons. J Physiol. 1976 Jul;259(2):427–443. doi: 10.1113/jphysiol.1976.sp011474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Henderson R., Ritchie J. M., Strichartz G. R. Evidence that tetrodotoxin and saxitoxin act at a metal cation binding site in the sodium channels of nerve membrane. Proc Natl Acad Sci U S A. 1974 Oct;71(10):3936–3940. doi: 10.1073/pnas.71.10.3936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Henderson R., Ritchie J. M., Strichartz G. R. The binding of labelled saxitoxin to the sodium channels in nerve membranes. J Physiol. 1973 Dec;235(3):783–804. doi: 10.1113/jphysiol.1973.sp010417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hille B. Charges and potentials at the nerve surface. Divalent ions and pH. J Gen Physiol. 1968 Feb;51(2):221–236. doi: 10.1085/jgp.51.2.221. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hille B. Pharmacological modifications of the sodium channels of frog nerve. J Gen Physiol. 1968 Feb;51(2):199–219. doi: 10.1085/jgp.51.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hille B. The receptor for tetrodotoxin and saxitoxin. A structural hypothesis. Biophys J. 1975 Jun;15(6):615–619. doi: 10.1016/S0006-3495(75)85842-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Hille B., Woodhull A. M., Shapiro B. I. Negative surface charge near sodium channels of nerve: divalent ions, monovalent ions, and pH. Philos Trans R Soc Lond B Biol Sci. 1975 Jun 10;270(908):301–318. doi: 10.1098/rstb.1975.0011. [DOI] [PubMed] [Google Scholar]
  15. Keynes R. D., Bezanilla F., Taylor R. E., Rojas E. The rate of action of tetrodotoxin on sodium conductance in the squid giant axon. Philos Trans R Soc Lond B Biol Sci. 1975 Jun 10;270(908):365–375. doi: 10.1098/rstb.1975.0016. [DOI] [PubMed] [Google Scholar]
  16. Krueger B. K., Ratzlaff R. W., Strichartz G. R., Blaustein M. P. Saxitoxin binding to synaptosomes, membranes, and solubilized binding sites from rat brain. J Membr Biol. 1979 Nov 30;50(3-4):287–310. doi: 10.1007/BF01868894. [DOI] [PubMed] [Google Scholar]
  17. McLaughlin S. G., Szabo G., Eisenman G. Divalent ions and the surface potential of charged phospholipid membranes. J Gen Physiol. 1971 Dec;58(6):667–687. doi: 10.1085/jgp.58.6.667. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Pooler J. P., Valenzeno D. P. Titration of sodium channel sites for hydrogen ion block and sensitized photochemical modification of lobster axons. Biochim Biophys Acta. 1979 Aug 7;555(2):307–315. doi: 10.1016/0005-2736(79)90170-6. [DOI] [PubMed] [Google Scholar]
  19. Ritchie J. M., Rogart R. B., Strichartz G. R. A new method for labelling saxitoxin and its binding to non-myelinated fibres of the rabbit vagus, lobster walking leg, and garfish olfactory nerves. J Physiol. 1976 Oct;261(2):477–494. doi: 10.1113/jphysiol.1976.sp011569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Schauf C. L., Davis F. A. Sensitivity of the sodium and potassium channels of Myxicola giant axons to changes in external pH. J Gen Physiol. 1976 Feb;67(2):185–195. doi: 10.1085/jgp.67.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Schwarz J. R., Ulbricht W., Wagner H. H. The rate of action of tetrodotoxin on myelinated nerve fibres of Xenopus laevis and Rana esculenta. J Physiol. 1973 Aug;233(1):167–194. doi: 10.1113/jphysiol.1973.sp010304. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sevcik C. Binding of tetrodotoxin to squid nerve fibers. Two kinds of receptors? J Gen Physiol. 1976 Jul;68(1):95–103. doi: 10.1085/jgp.68.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sigworth F. J., Spalding B. C. Chemical modification reduces the conductance of sodium channels in nerve. Nature. 1980 Jan 17;283(5744):293–295. doi: 10.1038/283293a0. [DOI] [PubMed] [Google Scholar]
  24. Ulbricht W., Wagner H. H. The influence of pH on equilibrium effects of tetrodotoxin on myelinated nerve fibres of Rana esculenta. J Physiol. 1975 Oct;252(1):159–184. doi: 10.1113/jphysiol.1975.sp011139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weigele J. B., Barchi R. L. Analysis of saxitoxin binding in isolated rat synaptosomes using a rapid filtration assay. FEBS Lett. 1978 Jul 15;91(2):310–314. doi: 10.1016/0014-5793(78)81199-5. [DOI] [PubMed] [Google Scholar]

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