Alpha-Bungarotoxin binding to human muscle acetylcholine receptor: measurement of affinity, delineation of AChR subunit residues crucial to binding, and protection of AChR function by synthetic peptides.
Alpha-Bungarotoxin (alpha-BuTx) binds with high affinity to the nicotinic acetylcholine receptor (AChR) of most species, mainly to sequences around the two cysteines at positions 192 and 193 of the alpha-subunit, but other sequences of the alpha-subunit and of the adjacent gamma- or epsilon- and delta-subunits are also important in the native molecule. Alpha-BuTx binds strongly to human AChR but the short alpha neurotoxins, for instance Erabutoxin B, are relatively ineffective at the human neuromuscular junction. In this article we compare the affinity of 125I-alpha-BuTx for Torpedo and human muscle AChR and the ability of neurotoxins to inhibit this binding. We examine the contribution to alpha-BuTx binding of the three amino acids that differ between human and Torpedo AChR alpha-185-196. In addition, we show that an alpha-185-199, peptide that binds strongly to 125I-alpha-BuTx and can inhibit its binding in solution, is also capable of protecting the AChR on a cell line or at the neuromuscular junction. Such peptides might be useful in the treatment of acute envenoming or the autoantibody-mediated block of AChR function that can occur in human disorders.
Neurosciences Group, Institute of Molecular Medicine, John Radcliffe Hospital, Oxford, UK.
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Vincent
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Jacobson
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Curran
D000818:Animals; D002038:Bungarotoxins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D003039:Cobra Neurotoxin Proteins / Q000378:metabolism; D004870:Erabutoxins / Q000378:metabolism; D006801:Humans; D051379:Mice; D009132:Muscles / Q000378:metabolism*; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism; D011950:Receptors, Cholinergic / Q000378:metabolism*; D014101:Torpedo; D014407:Tumor Cells, Cultured
D002038:Bungarotoxins; D003039:Cobra Neurotoxin Proteins; D010455:Peptides; D011950:Receptors, Cholinergic; D004870:Erabutoxins; C025154:alpha-cobratoxin