Poster 16 - A comparative characterization of Bothrops jararacussu, Bothrops alternatus, Crotalus

Poster 16. Congresso da Sociedade Brasileira de Toxinologia, 8., Symposium of the Pan American Section of the International Society on Toxinology, 8., 2004, Angra dos Reis, Brasil. Abstracts... J. Venom. Anim. Toxins incl.Trop. Dis., 2004, 10, 3, p.375.

A comparative characterization of Bothrops jararacussu, Bothrops alternatus, Crotalus durissus terrificus (var. white) and Crotalus durissus collilineatus PLA2 by MALDI-TOF Mass Spectrometry and neurotoxic activity.

Bonfim, V.L., Ponce-Soto, L.A., Fagundes F.H.R., Damico, D.C.S., and Marangoni. S.

A comparative molecular mass profile characterization by matrix assisted laser desorption/ionization time of flight mass spectrometry (Maldi-Tof-MS) and neurotoxic activity on mouse phrenic nerve-diaphragm preparation of the neurotoxins from Bothropsjararacussu (Bj-IV, V and VII), Bothropsalternatus (BtII-2), Crotalusdurissus terrificus (var. white) (F15, F16 and F17) and Crotalusdurissus collilineatus (F6) was realized with the objective of establishing likeness and/or differences in relation to the neurotoxic between bothropics PLA2 and crotalics. Maldi-Tof-MS were performed using a Voyager-DE PRO MALDI-TOF mass spectrometry (Applied). 1mL of the sample matrix-solution, 2 mL. of neurotoxin PLA2 in a-ciano-4-hydroxycinnamic acid (a-CHBA) or sinapinic acid and their mass analyzed under the following conditions: accelerate voltage 25kV, laser fixed in 2890 mJ/com2, delay 300 nsec and in linear analysis mode. The neurotoxic study was realized through indirectly stimulated on mouse phrenic nerve-diaphragm preparation. This method was prepared as originally described by Bülbring (1946). Comparative molecular mass showed a profile with some differences between the PLA2 studied in relation of the results determined by MALDI-TOF and the neurotoxic effect. The molecular masses of the PLA2 were in the ranges of 13898,71 and 14,532,76 Da (Bothrops and Crotalus). The neurotoxic activity of the PLA2 studied showed ability to block neuromuscular transmission at a concentration of 50 ug/ml, for example, the Bj-IV caused a 50% block at a 11,38±6.00 min, the Bj-V at a 38,69±3,15 min and Bj-VII at a 39,18±7,17 min. None of the other crotalics venoms studied here caused complete neuromuscular blockade (p<0,05). Comparative molecular mass MALDI-TOF profile and the blocked time on neuromuscular transmission suggest that those differences of mass contribution with the difference in the respective times of blocked. The absence of neuromuscular transmission blocked could be due the presence of specific domains in the bothropics PLA2, which could be related with the mass too.