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 μM) 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 p A 2 of 8.36±0.17. Acantoxin IVa was approximately two-fold less potent than the long-chain (Type II) neurotoxin, α-bungarotoxin. With a p K i value of 4.48, acantoxin IVa was approximately 25,000 times less potent than α-bungarotoxin at α7-type neuronal nAChR. However, in contrast to α-bungarotoxin, acantoxin IVa completely inhibited specific [ 3 H ]-methyllycaconitine (MLA) binding in rat hippocampus homogenate. Acantoxin IVa had no activity at ganglionic nAChR, α4β2 subtype neuronal nAChR or cytisine-resistant [ 3 H ]-epibatidine binding sites. While long-chain neurotoxin resistant [ 3 H ]-MLA binding in hippocampus homogenate requires further investigation, we have shown that a short-chain (Type I) neurotoxin is capable of fully inhibiting specific [ 3 H ]-MLA binding.