A single-step purification of bothropstoxin-1

Bothrops venoms are complex mixtures of components with a wide range of biological activities. Among these substances, myotoxins have been investigated by several groups. Bothropstoxin-1 (Bthtx-1) is a phospholipase A2-like basic myotoxin from Bothrops jararacussu. The purification of this component involves two chromatographic steps. Although providing a pure material, the association of these two steps is time consuming and a single-step method using high performance chromatography media would be useful. In the present study, we describe a single-step purification method for Bthtx-1. Bothrops jararacussu venom was dissolved in 1 ml buffer. After centrifugation, the supernatant was injected into a Resource-S cation exchange column connected to an FPLC system and eluted with a linear salt gradient. The complete procedure took 20 min, representing a considerable time gain when compared to a previously described method (Homsi-Brandenburgo MI et al. (1988) Toxicon, 26: 615-627). Bthtx-1 purity and identity, assessed by SDS-PAGE and N-terminal sequencing, resulted in a single band with a molecular mass of about 14 kDa and the expected sequence of the first 5 residues, S-L-F-E-L. Although the amount of protein purified after each run is lower than in the previously described method, we believe that this method may be useful for small-scale purifications

The in vitro and in vivo pharmacological activity of Boiga dendrophila (mangrove catsnake) venom

10.1111/j.1474-8673.2004.00322.xAutonomic and Autacoid Pharmacology244107-113AAPU

Caracterização individual do veneno de Bothrops alternatus Duméril, Bibron & Duméril em função da distribuição geográfica no Brasil (Serpentes,Viperidae) Individual characterization of Bothrops alternatus Duméril, Bibron & Duméril venoms, according to their geographic distribution in Brazil (Serpentes, Viperidae)

Bothrops alternatus Duméril, Bibron & Duméril, 1854 é uma serpente de importância em saúde pública, com ampla distribuição geográfica, desde o Mato Grosso do Sul até o sudeste do Brasil, chegando até a Argentina e Uruguai, ocupando vários domínios morfoclimáticos. Neste trabalho investigou-se a variação do veneno de adultos de Bothrops alternatus, em função de sua distribuição geográfica no Brasil, comparativamente ao veneno elaborado sob a forma de "pool" desta espécie (veneno referência), que inclui serpentes, em sua maioria, da região do estado de São Paulo. Foram analisadas as atividades letal, coagulante sobre o plasma, proteolítica sobre a caseína e miotóxica, bem como os padrões eletroforéticos de 61 amostras individuais de veneno contrapostas ao "pool". Os resultados mostraram que o veneno de B. alternatus é pouco ativo, comparativamente ao de outros Bothrops Wagler, 1824. A variação individual prevaleceu, não apresentando correlação com as áreas de distribuição geográfica e domínios morfoclimáticos, porém a atividade coagulante das amostras de veneno provenientes do nordeste da distribuição geográfica apresentaram-se menos ativas comparativamente às da porção central da distribuição. Os venenos provenientes das bordas da distribuição apresentaram ações proteolíticas e miotóxicas mais intensas, que estatisticamente não foram significativamente diferentes. As variações individuais prevaleceram.<br>Bothrops alternatus Duméril, Bibron & Duméril, 1854 snakebites are an important public health problem in Brazil. Such snakes are found from Mato Grosso do Sul (central Brazil) to southeastern Brazil, reaching even Argentina and Uruguay and thereby occupying different morphoclimatic domains. This work investigated venom variation occurring in adult specimens of B. alternatus specimens, according to their geographic distribution in Brazil. The standard venom pool (reference venom) produced by Instituto Butantan, which includes mostly venoms from B. alternatus specimens captured in São Paulo State, was also used for comparison. Lethal, myotoxic, proteolytic (on casein), and coagulant (on human plasma) activities, as well as the electrophoretic patterns of 61 individual venom samples, were evaluated and compared to those of the reference venom pool. Results showed that B. alternatus venom activities are less potent than those of other Bothrops Wagler, 1824 species. A great individual variation was noticed, which could not be correlated with either geographic distribution or morphoclimatic domains. Coagulant activity of venom samples obtained from specimens captured in the northeastern area of B. alternatus distribution was less intense if compared to those obtained from the central zone. Venoms from the peripheric area of B. alternatus distribution presented higher proteolytic and myotoxic activities, but no statistically significant difference was observed. In summary, individual variation predominated over geographic distribution