A powerful inhibition of mammalian acetylcholinesterase was detected in the venom of the snake Dendroaspis angusticeps (green mamba). The substances responsible for such inhibition were isolated and purified by gel filtration on Sephadex G-50 and ion exchange chromatography on Bio-Rex 70 and SP Sephadex C-25. These substances were polypeptides and were named, fasciculins. Upon intraperitoneal injection into mice fasciculins elicited severe, generalized, long-lasting muscle fasciculations with complete clinical recovery. In vitro preincubation with fasciculins at concentrations of 0.01 ?g ml(?1) inhibited brain and muscle acetylcholinesterases up to 80%. Histochemical assay for acetylcholinesterase showed an almost complete disappearance of the black-brown precipitate at the neuromuscular end-plate after in vitro incubation with fasciculins. Fasciculins represent a new type of acetylcholinesterase inhibitors provoking muscle fasciculations through a powerful inhibition of enzyme activity at the neuromuscular end-plate, interfering with the normal degradative activity of the acetylcholine molecule. Fasciculins are also powerful inhibitors of brain acetylcholinesterases.