Document Detail


Characterization of curaremimetic neurotoxin binding sites on membrane fractions derived from the human medulloblastoma clonal line, TE671.
MedLine Citation:
PMID:  3701338     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Studies were conducted on curaremimetic neurotoxin binding to the nicotinic acetylcholine receptor present on membrane fractions derived from the human medulloblastoma clonal line, TE671. High-affinity binding sites (KD = 2 nM for 1-h incubation at 20 degrees C) and low-affinity binding sites (KD = 40 nM) for 125I-labeled alpha-bungarotoxin are present in equal quantities (60 fmol/mg membrane protein). The kinetically determined dissociation constant for high-affinity binding of toxin is 0.56 nM (k1 = 6.3 X 10(-3) min-1 nM-1; k-1 = 3.5 X 10(-3) min-1) at 20 degrees C. Nicotine, d-tubocurarine, and acetylcholine are among the most effective inhibitors of high-affinity toxin binding. The quantity of toxin binding sites and their affinity for cholinergic agonists is sensitive to reduction, alkylation, and/or oxidation of membrane sulfhydryl residues. High-affinity toxin binding sites that have been subjected to reaction with the sulfhydryl reagent dithiothreitol are irreversibly blocked by the nicotinic receptor affinity reagent bromoacetylcholine. High-affinity toxin binding is inhibited in the presence of either of two polyclonal antisera or a monoclonal antibody raised against nicotinic acetylcholine receptors from fish electric tissue. Taken together, these results indicate that curaremimetic neurotoxin binding sites on membrane fractions of the TE671 cell line share some properties with nicotinic acetylcholine receptors of peripheral origin and with toxin binding sites on other neuronal tissues.
Authors:
R J Lukas
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Journal of neurochemistry     Volume:  46     ISSN:  0022-3042     ISO Abbreviation:  J. Neurochem.     Publication Date:  1986 Jun 
Date Detail:
Created Date:  1986-06-20     Completed Date:  1986-06-20     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  2985190R     Medline TA:  J Neurochem     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1936-41     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Acetylcholine / analogs & derivatives,  metabolism,  pharmacology
Binding Sites
Binding, Competitive
Bungarotoxins / metabolism
Cell Line
Clone Cells / metabolism*
Curare / metabolism*
Dithiothreitol / pharmacology
Ethylmaleimide / pharmacology
Humans
Kinetics
Medulloblastoma / metabolism*
Nicotine / metabolism
Receptors, Cholinergic / metabolism
Tubocurarine / metabolism
Grant Support
ID/Acronym/Agency:
NS-16821/NS/NINDS NIH HHS; S07 RR05872/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Bungarotoxins; 0/Receptors, Cholinergic; 128-53-0/Ethylmaleimide; 3483-12-3/Dithiothreitol; 51-84-3/Acetylcholine; 54-11-5/Nicotine; 57-95-4/Tubocurarine; 8063-06-7/Curare

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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