Structural And Neurotoxic Properties Of Crotoxine-like In Chick Biventer Cervicis Nerve-Muscle Preparation, Isolated From Crotalus durissus collilineatus Venom.

¹de Santis, L.H., ¹Ponce-Soto, L.A., ¹Fagundes, F.H.R., ²Leite, G. B., ²Rodrigues-Simioni, L. and ¹Marangoni, S.

1 Departamento de Bioquímica, Instituto de Biologia (IB) and 3 Departamento de Farmacologia, Faculdade de Ciências Médicas, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil

The crotoxine-like from Crotalusdurissus collilineatus was purified using molecular exclusion in a HPLC system, we find one basic PLA2 of 14 kDa and two crotapotins isoforms, F3 of 8 kDa and F4 of 9 kDa in agreement to the analysis of amino acids and the analysis electrophoretic in SDS PAGE. The molecular mass was confirmed by MALDI-TOF mass spectrometry. The accurate mass gotten for PLA2 was 14 943.18 Da, for F3 was 8 693.24 Da and for F4 was 9 341.56 Da. The effect of the crotoxine-like of Crotalus durissus collilineatus was investigated in the preparation biventer cervicis chick nerve-muscle. The assay was carried out for indirect stimulus, and was observed the effect neurotoxic of the crotoxine-like which is a potent blocking agent of the contractile answer. The crotoxine-like was used in 1, 5, 10 and 20 mg/ml, being the minimum dose of 1 mg/ml to become the irreversible blockade. The presence of acetylcholine (10 and 20 mg/ml) and KCl (13.4 mM) before and after adding the crotoxine-like not showed a reduction of the contractile answer. We can establish that the crotoxine-like of Crotalusdurissus collilineatus is a potent presynaptic neurotoxin able to block the neuromuscular transmission. The other conclusion was that the crotoxin-like have an effect of irreversible blockade as well as is dose-dependent.

Financial Support: CAPES and FAPESP

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