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Nicotinic acetylcholine receptor contains multiple binding sites: Evidence from binding of α-dendrotoxin

Conti-Tronconi, Bianca M. and Raftery, Michael A. (1986) Nicotinic acetylcholine receptor contains multiple binding sites: Evidence from binding of α-dendrotoxin. Proceedings of the National Academy of Sciences of the United States of America, 83 (17). pp. 6646-6650. ISSN 0027-8424. PMCID PMC386561. doi:10.1073/pnas.83.17.6646.

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We have studied the stoichiometry of the binding of the long α-neurotoxins from the venom of Dendroaspis viridis (α-dendrotoxin) and Naja naja siamensis (α-cobratoxin) to the membrane-bound acetylcholine receptor (AcChoR) from Torpedo californica electric organ. The number of toxin molecules bound to one AcChoR molecule was determined by simultaneous-quantitative gas-phase microsequencing of all the amino acid sequences present in AcChoR-α-neurotoxin complexes. This method permits the use of homogeneous (nonradiolabeled) preparations of native toxins to obtain molar ratios of neurotoxin-receptor complexes. The stoichiometry obtained for α-cobratoxin was 2.1 ± 0.2 (n = 4), in agreement with the accepted view that α-cobratoxin, like α-bungarotoxin, binds to the two α subunits, which are constituent polypeptides of the AcChoR molecule. α-Dendrotoxin gave a stoichiometry of 4.1 ± 0.5 (n = 12); therefore, the AcChoR molecule contains four binding sites for this α-neurotoxin, two of which are recognized by α-cobratoxin. In support of this contention we have also found that when the AcChoR is saturated with α-bungarotoxin, addition of α-dendrotoxin markedly accelerates the dissociation of the bound α-bungarotoxin, demonstrating that the occupancy of the additional two sites by the latter toxin influences and decreases the affinity of the former toxin for its two binding sites. The fact that the AcChoR molecule is a pseudoxymmetric complex of five highly homologous peptides suggests the possibility that as many as five binding sites for cholinergic ligand could be present, one on each subunit.

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Additional Information:© 1986 by the National Academy of Sciences. Communicated by James Bonner, March 14, 1986. This research was supported by U.S. Public Health Service Grant NS10294 and U.S. Army Research Office Contract DAMD17-82-C-2175. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.
Funding AgencyGrant Number
Army Research Office (ARO)DAMD17-82-C-2175
Issue or Number:17
PubMed Central ID:PMC386561
Record Number:CaltechAUTHORS:CONpnas86b
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:7765
Deposited By: Tony Diaz
Deposited On:30 Jul 2007
Last Modified:08 Nov 2021 20:45

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