Lophotoxin is a slow binding irreversible inhibitor of nicotinic acetylcholine receptors.
Lophotoxin and the bipinnatins are members of the lophotoxin family of marine neurotoxins, which covalently react with Tyr190 in the alpha-subunits of the nicotinic acetylcholine receptor. Bipinnatin-A, -B, and -C are protoxins that have been shown to spontaneously convert from inactive to active toxins during preincubation in buffer. However, in this report, we show that preincubation of lophotoxin did not result in an increase in the subsequent rate of irreversible inhibition of nicotinic receptors. Thus, unlike the bipinnatins, lophotoxin does not appear to be an inactive protoxin. Lophotoxin preferentially inhibited one of the two acetylcholine-binding sites on the receptor, and this preference resulted from both a higher reversible affinity and a faster rate of irreversible inhibition at this site. Association of 125I-alpha-bungarotoxin in the presence of lophotoxin was analyzed to obtain the apparent reversible association and dissociation rate constants for lophotoxin. The apparent association rate constant of lophotoxin was approximately 10(6)-fold slower than expected for a diffusion-limited interaction, indicating that lophotoxin is a slow binding irreversible inhibitor. The kinetic constants that describe the interaction of lophotoxin with the receptor did not change in the presence of dibucaine, suggesting that, unlike agonists, the slow apparent association of lophotoxin does not result from a slow transition of the receptor to a desensitized conformation.
Groebe
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Department of Pharmacology, School of Medicine, University of Pittsburgh, Pennsylvania 15261.
Abramson
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D002038:Bungarotoxins / Q000378:metabolism; D002478:Cells, Cultured; D007457:Iodine Radioisotopes; D007700:Kinetics; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic / Q000378:metabolism; D013729:Terpenes / Q000378:metabolism / Q000494:pharmacology*
D002038:Bungarotoxins; D007457:Iodine Radioisotopes; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic; D013729:Terpenes; C030303:lophotoxin