title: Isolation and characterization at cholinergic nicotinic receptors of a neurotoxin from the venom of the Acanthophis sp. Seram death adder.
abstract: The present study describes the isolation of the first neurotoxin (acantoxin IVa) from Acanthophis sp. Seram death adder venom and an examination of its activity at nicotinic acetylcholine receptor (nAChR) subtypes. Acantoxin IVa (MW 6815; 0.1-1.0 microM) caused concentration-dependent inhibition of indirect twitches (0.1 Hz, 0.2 ms, supramaximal V) and inhibited contractile responses to exogenous nicotinic agonists in the chick biventer cervicis nerve-muscle, confirming that this toxin is a postsynaptic neurotoxin. Acantoxin IVa (1-10 nM) caused pseudo-irreversible antagonism at skeletal muscle nAChR with an estimated pA2 of 8.36+/-0.17. Acantoxin IVa was approximately two-fold less potent than the long-chain (Type II) neurotoxin, alpha-bungarotoxin. With a pKi value of 4.48, acantoxin IVa was approximately 25,000 times less potent than alpha-bungarotoxin at alpha7-type neuronal nAChR. However, in contrast to alpha-bungarotoxin, acantoxin IVa completely inhibited specific [3H]-methyllycaconitine (MLA) binding in rat hippocampus homogenate. Acantoxin IVa had no activity at ganglionic nAChR, alpha4beta2 subtype neuronal nAChR or cytisine-resistant [3H]-epibatidine binding sites. While long-chain neurotoxin resistant [3H]-MLA binding in hippocampus homogenate requires further investigation, we have shown that a short-chain (Type I) neurotoxin is capable of fully inhibiting specific [3H]-MLA binding.
authors.affiliation: Department of Pharmacology, Monash University, Victoria 3800, Australia.
authors.forename: Janith C
authors.initials: JC
authors.lastname: Wickramaratna
authors.forename: Bryan G
authors.initials: BG
authors.lastname: Fry
authors.forename: Richard E
authors.initials: RE
authors.lastname: Loiacono
authors.forename: Marie-Isabel
authors.initials: MI
authors.lastname: Aguilar
authors.forename: Paul F
authors.initials: PF
authors.lastname: Alewood
authors.forename: Wayne C
authors.initials: WC
authors.lastname: Hodgson
mesh_terms: D000818:Animals; D001665:Binding Sites; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000494:pharmacology; D002642:Chick Embryo; D003553:Cystine / Q000494:pharmacology; D004546:Elapid Venoms / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology*; D017815:Elapidae; D005260:Female; D045888:Ganglion Cysts / Q000378:metabolism; D006168:Guinea Pigs; D008970:Molecular Weight; D018482:Muscle, Skeletal / Q000378:metabolism; D009498:Neurotoxins / Q000633:toxicity; D018733:Nicotinic Antagonists / Q000633:toxicity*; D010455:Peptides / Q000302:isolation & purification / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011978:Receptors, Nicotinic / Q000378:metabolism*; D020539:Sequence Analysis, Protein; D014316:Tritium; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: D019086:Bridged Bicyclo Compounds, Heterocyclic; C579165:Chrna7 protein, rat; D004546:Elapid Venoms; D009498:Neurotoxins; D018733:Nicotinic Antagonists; D010455:Peptides; D011725:Pyridines; D011978:Receptors, Nicotinic; C488186:acantoxin IVa, Acanthophis; D064569:alpha7 Nicotinic Acetylcholine Receptor; C403531:nicotinic receptor alpha4beta2; D014316:Tritium; D003553:Cystine; C082748:epibatidine