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Research ArticleArticle

Characterization of Fragment C and Tetanus Toxin Binding to Rat Brain Membranes

RONALD L. GOLDBERG, TOMMASO COSTA, WILLIAM H. HABIG, LEONARD D. KOHN and M. CAROLYN HARDEGREE
Molecular Pharmacology November 1981, 20 (3) 565-570;
RONALD L. GOLDBERG
Bacterial Toxins Branch, Division of Bacterial Products, Bureau of Biologics, Food and Drug Administration, Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, and Laboratory of Biochemical Pharmacology, National Institute of Arthritis, Metabolism and Digestive Diseases, Bethesda, Maryland 20205
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TOMMASO COSTA
Bacterial Toxins Branch, Division of Bacterial Products, Bureau of Biologics, Food and Drug Administration, Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, and Laboratory of Biochemical Pharmacology, National Institute of Arthritis, Metabolism and Digestive Diseases, Bethesda, Maryland 20205
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WILLIAM H. HABIG
Bacterial Toxins Branch, Division of Bacterial Products, Bureau of Biologics, Food and Drug Administration, Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, and Laboratory of Biochemical Pharmacology, National Institute of Arthritis, Metabolism and Digestive Diseases, Bethesda, Maryland 20205
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LEONARD D. KOHN
Bacterial Toxins Branch, Division of Bacterial Products, Bureau of Biologics, Food and Drug Administration, Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, and Laboratory of Biochemical Pharmacology, National Institute of Arthritis, Metabolism and Digestive Diseases, Bethesda, Maryland 20205
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M. CAROLYN HARDEGREE
Bacterial Toxins Branch, Division of Bacterial Products, Bureau of Biologics, Food and Drug Administration, Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, and Laboratory of Biochemical Pharmacology, National Institute of Arthritis, Metabolism and Digestive Diseases, Bethesda, Maryland 20205
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Abstract

The binding characteristics of 125I-labeled tetanus toxin and 125I-labeled fragment C to rat brain membranes have been studied. Fragment C is a proteolytically derived portion of the holotoxin. Tetanus toxin is slightly more potent than fragment C in displacing 125I-labeled tetanus toxin bound to rat brain membranes. Conversely, fragment C is more effective than toxin in displacing bound 125I-labeled fragment C. However, the binding curves for the two compounds are in both cases parallel, suggesting that they compete for the same site. The equilibrium affinity constants for the binding of tetanus toxin and fragment C as obtained by both direct and indirect binding experiments were found to be in the nanomolar range (2-12 nM). This relatively high affinity is in contrast with the large number of binding sites (estimated as 2-9 nmoles/mg of protein). This concentration is consistent with the reported content of the gangliosides, GT1 and GD1b, in rat brain. The specificity of the toxin binding site has been tested; a number of putative neurotransmitters as well as inhibitors of axonal transport were unable to affect the binding at concentrations several orders of magnitude higher than the toxin or fragment C. The anticonvulsant drug diazepam, reported useful in the treatment of human tetanus, was equally inactive in displacing 125I-labeled tetanus toxin binding. The possible pharmacological relevance of the binding site for tetanus toxin and this fragment is discussed.

  • Copyright © 1981 by The American Society for Pharmacology and Experimental Therapeutics

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Molecular Pharmacology
Vol. 20, Issue 3
1 Nov 1981
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Research ArticleArticle

Characterization of Fragment C and Tetanus Toxin Binding to Rat Brain Membranes

RONALD L. GOLDBERG, TOMMASO COSTA, WILLIAM H. HABIG, LEONARD D. KOHN and M. CAROLYN HARDEGREE
Molecular Pharmacology November 1, 1981, 20 (3) 565-570;

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Research ArticleArticle

Characterization of Fragment C and Tetanus Toxin Binding to Rat Brain Membranes

RONALD L. GOLDBERG, TOMMASO COSTA, WILLIAM H. HABIG, LEONARD D. KOHN and M. CAROLYN HARDEGREE
Molecular Pharmacology November 1, 1981, 20 (3) 565-570;
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