α-Bungarotoxin-ferritin conjugates were used to visualize by freeze-fracture and thin-section electron microscopy toxin-binding sites, presumably acetylcholine (ACh) receptors, in membranes of muscle cells grown in tissue culture. Toxin conjugated to ferritin by a glutaraldehyde reaction and purified by column chromatography in a buffer of high ionic strength remains active in blocking the effect of iontophoretically applied ACh. The potency of the conjugates was decreased 5-10 times compared to native α-bungarotoxin. Toxin-ferritin conjugates were identified in small clusters which were not uniformly distributed over the surface membrane. Binding was inhibited by preincubation in D-tubocurare or unconjugated toxin. The relation of the clusters to the non-uniform distribution of ACh sensitivity and α-bungarotoxin binding on cultured muscle fibres is discussed.
JOURNAL ARTICLE|
01 November 1974
Acetylcholine Receptors of Cultured Muscle Cells Demonstrated with Ferritin-α-Bungarotoxin Conjugates
B. T. HOURANI,
B. T. HOURANI
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: University Hospital, San Diego, California, U.S.A.
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B. F. TORAIN,
B. F. TORAIN
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.
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M. P. HENKART,
M. P. HENKART
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.
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R. L. CARTER,
R. L. CARTER
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.
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V. T. MARCHESI,
V. T. MARCHESI
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Yale University School of Medicine, New Haven, Connecticut, U.S.A.
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G. D. FISCHBACH
G. D. FISCHBACH
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Harvard Medical School, Boston, Massachusetts, U.S.A.
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B. T. HOURANI
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: University Hospital, San Diego, California, U.S.A.
B. F. TORAIN
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.
M. P. HENKART
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.
R. L. CARTER
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.
V. T. MARCHESI
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Yale University School of Medicine, New Haven, Connecticut, U.S.A.
G. D. FISCHBACH
Laboratory of Experimental Pathology, National Institute of Arthritis, Metabolism, and Digestive Diseases, and Behavioural Biology Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, Maryland 20014, U.S.A.; Present address: Harvard Medical School, Boston, Massachusetts, U.S.A.
Received:
29 Mar 1974
Online ISSN: 1477-9137
Print ISSN: 0021-9533
Copyright © 1974 The Company of Biologists Ltd.
1974
J Cell Sci (1974) 16 (2): 473–479.
Article history
Received:
29 Mar 1974
Citation
B. T. HOURANI, B. F. TORAIN, M. P. HENKART, R. L. CARTER, V. T. MARCHESI, G. D. FISCHBACH; Acetylcholine Receptors of Cultured Muscle Cells Demonstrated with Ferritin-α-Bungarotoxin Conjugates. J Cell Sci 1 November 1974; 16 (2): 473–479. doi: https://doi.org/10.1242/jcs.16.2.473
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