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. 1979 Jun;76(6):2576–2579. doi: 10.1073/pnas.76.6.2576

Specific binding of perhydrohistrionicotoxin to Torpedo acetylcholine receptor.

J Elliott, S M Dunn, S G Blanchard, M A Raftery
PMCID: PMC383650  PMID: 288047

Abstract

Torpedo californica postsynaptic membrane fragments were treated with base, which resulted in membranes that were depleted of many nonacetylcholine receptor polypeptides and contained acetylcholine receptor subunits of Mr 40,000, 50,000, 60,000, and 65,000 (Raftery, M.A., Vandlen, R.L., Reed, K.L. & Lee T. (1975) Cold Spring Harbor Symp. Quant. Biol. 40, 193-202). A 43,000-Mr polypeptide and some other components were quantitatively extracted. Base-treated membranes retained the capacity to bind [3H]perhydrohistrionicotoxin and the local anesthetics dibucaine and tetracaine. The regulation of this binding by carbamylcholine, as well as the kinetic mechanism of perhydrohistrionicotoxin binding, was unchanged. [3H]Perhydrohistrionicotoxin binding activity was largely reconstituted from 2% sodium cholate extracts of base-treated membranes. Therefore, the perhydrostrionicotoxin binding site appears to be located on one or more of the acetylcholine receptor polypeptides, and the reconstitution of that binding site from detergent extracts does not require the presence of a 43,000-Mr polypeptide.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Albuquerque E. X., Barnard E. A., Chiu T. H., Lapa A. J., Dolly J. O., Jansson S. E., Daly J., Witkop B. Acetylcholine receptor and ion conductance modulator sites at the murine neuromuscular junction: evidence from specific toxin reactions. Proc Natl Acad Sci U S A. 1973 Mar;70(3):949–953. doi: 10.1073/pnas.70.3.949. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aratani M., Dunkerton L. V., Fukuyama T., Kishi Y., Kakoi H., Sugiura S., Inoue S. Letter: Synthetic studies on histrionicotoxins. I. A stereocontrolled synthesis of (plus or minus)-perhydrohistrionicotoxin. J Org Chem. 1975 Jun 27;40(13):2009–2011. doi: 10.1021/jo00901a037. [DOI] [PubMed] [Google Scholar]
  3. Burgermeister W., Catterall W. A., Witkop B. Histrionicotoxin enhances agonist-induced desensitization of acetylcholine receptor. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5754–5758. doi: 10.1073/pnas.74.12.5754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dolly J. O., Albuquerque E. X., Sarvey J., Mallick B., Barnard E. A. Binding of perhydro-histrionicotoxin to the postsynaptic membrane of skeletal muscle in relation to its blockage of acetylcholine-induced depolarization. Mol Pharmacol. 1977 Jan;13(1):1–14. [PubMed] [Google Scholar]
  5. Eldefrawi A. T., Eldefrawi M. E., Albuquerque E. X., Oliveira A. C., Mansour N., Adler M., Daly J. W., Brown G. B., Burgermeister W., Witkop B. Perhydrohistrionicotoxin: a potential ligand for the ion conductance modulator of the acetylcholine receptor. Proc Natl Acad Sci U S A. 1977 May;74(5):2172–2176. doi: 10.1073/pnas.74.5.2172. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Elliot J., Raftery M. A. Interactions of perhydrohistrionicotoxin with postsynaptic membranes. Biochem Biophys Res Commun. 1977 Aug 22;77(4):1347–1353. doi: 10.1016/s0006-291x(77)80127-7. [DOI] [PubMed] [Google Scholar]
  7. Fukuyama T., Dunkerton L. V., Aratani M., Kishi Y. Letter: Synthetic studies on histrionicotoxins. II. A practical synthetic route to (plus or minus)-perhydro-and (plus or minus)-octahydrohistrionicotoxin. J Org Chem. 1975 Jun 27;40(13):2011–2012. doi: 10.1021/jo00901a038. [DOI] [PubMed] [Google Scholar]
  8. Karlin A., Weill C. L., McNamee M. G., Valderrama R. Facets of the structures of acetylcholine receptors from Electrophorus and Torpedo. Cold Spring Harb Symp Quant Biol. 1976;40:203–210. doi: 10.1101/sqb.1976.040.01.022. [DOI] [PubMed] [Google Scholar]
  9. Kato G., Changeux J. P. Studies on the effect of histrionicotoxin on the monocellular electroplax from Electrophorus electricus and on the binding of (3H)acetylcholine to membrane fragments from Torpedo marmorata. Mol Pharmacol. 1976 Jan;12(1):92–100. [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Martinez-Carrion M., Raftery M. A. Use of a fluorescent probe for the study of ligand binding by the isolated cholinergic receptor of Torpedo californica. Biochem Biophys Res Commun. 1973 Dec 19;55(4):1156–1164. doi: 10.1016/s0006-291x(73)80016-6. [DOI] [PubMed] [Google Scholar]
  13. Moody T., Schmidt J., Raftery M. A. Binding of acetylcholine and related compounds to purified acetylcholine receptor from Torpedo Californica electroplax. Biochem Biophys Res Commun. 1973 Aug 6;53(3):761–772. doi: 10.1016/0006-291x(73)90158-7. [DOI] [PubMed] [Google Scholar]
  14. Quast U., Schimerlik M., Lee T., Witzemann T. L., Blanchard S., Raftery M. A. Ligand-induced conformation changes in Torpedo californica membrane-bound acetylcholine receptor. Biochemistry. 1978 Jun 13;17(12):2405–2414. doi: 10.1021/bi00605a024. [DOI] [PubMed] [Google Scholar]
  15. Quast U., Schimerlik M., Raftery M. A. Stopped flow kinetics of carbamylcholine binding to membrane bound acetylcholine receptor. Biochem Biophys Res Commun. 1978 Apr 14;81(3):955–964. doi: 10.1016/0006-291x(78)91444-4. [DOI] [PubMed] [Google Scholar]
  16. Raftery M. A., Vandlen R. L., Reed K. L., Lee T. Characterization of Torpedo californica acetylcholine receptor: its subunit composition and ligand-binding properties. Cold Spring Harb Symp Quant Biol. 1976;40:193–202. doi: 10.1101/sqb.1976.040.01.021. [DOI] [PubMed] [Google Scholar]
  17. Schmidt J., Raftery M. A. A simple assay for the study of solubilized acetylcholine receptors. Anal Biochem. 1973 Apr;52(2):349–354. doi: 10.1016/0003-2697(73)90036-5. [DOI] [PubMed] [Google Scholar]
  18. Schmidt J., Raftery M. A. Purification of acetylcholine receptors from Torpedo californica electroplax by affinity chromatography. Biochemistry. 1973 Feb 27;12(5):852–856. doi: 10.1021/bi00729a011. [DOI] [PubMed] [Google Scholar]
  19. Sobel A., Heidmann T., Hofler J., Changeux J. P. Distinct protein components from Torpedo marmorata membranes carry the acetylcholine receptor site and the binding site for local anesthetics and histrionicotoxin. Proc Natl Acad Sci U S A. 1978 Jan;75(1):510–514. doi: 10.1073/pnas.75.1.510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Weill C. L., McNamee M. G., Karlin A. Affinity-labeling of purified acetylcholine receptor from Torpedo californica. Biochem Biophys Res Commun. 1974 Dec 11;61(3):997–1003. doi: 10.1016/0006-291x(74)90254-x. [DOI] [PubMed] [Google Scholar]

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