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Specific binding of perhydrohistrionicotoxin to Torpedo acetylcholine receptor

Elliott, Janet and Dunn, Susan M. J. and Blanchard, Steven G. and Raftery, Michael A. (1979) Specific binding of perhydrohistrionicotoxin to Torpedo acetylcholine receptor. Proceedings of the National Academy of Sciences of the United States of America, 76 (6). pp. 2576-2579. ISSN 0027-8424.

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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 perhydrohistrionicotoxin 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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Additional Information:© 1979 by the National Academy of Sciences. Communicated by James F. Bonner, February 28, 1979. H12-HTX and H8-HTX used in the preparation of [3H]H12-HTX were kind gifts of Dr. Y. Kishi, who produced them synthetically (28, 29). Thanks are due to John Racs for help with Torpedo membrane preparations and Valerie Purvis for typing the manuscript and drawing the figures. This research was supported by grants from the National Institutes of Health (U.S. Public Health Service NS 10294) and from the Muscular Dystrophy Association of America, as well as by National Institutes of Health Predoctoral Fellowships to J.E. and S.G.B. 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.
Subject Keywords:reconstitution; local anesthetics; cholinergic receptor; ion translocation; α-bungarotoxin sites
Issue or Number:6
Record Number:CaltechAUTHORS:ELLpnas79
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:8482
Deposited By: Tony Diaz
Deposited On:15 Aug 2007
Last Modified:02 Oct 2019 23:51

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