Purificação e caracterização de uma fosfolipase A2 do veneno amarelo de serpentes Crotalus durissus collilinetaus
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Data
2009-06-29
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Universidade Federal de Goiás
Resumo
In Brazil, the snake Crotalus durissus collilineatus is much studied, particularly its
component crotoxina. The crotoxina is the most toxic component of venom from the
species cascavéis amicanas Crotalus durissus. It is composed of two different subunits, the
crotapotina (acid component) and a phospholipase A2 (basic component). The enzyme
phospholipase A2 is the most studied of these poisons, being largely responsible for its
toxicity. Its main effects are neurotoxicity, myotoxicity, cytotoxicity, platelet aggregation,
anti-coagulant and bactericide. This study aims to purify and characterize a new
phospholipase A2 isolated from Crotalus durissus collilineatus, and uses it as a model for
studies of compounds with potential antinflamatório. Samples (5mg) of crude venom of
Crotalus durissus collilineatus were applied on a C18 column coupled to a system
semipreparativa HPLC. 20 peaks were obtained for proteins / peptides, and the increased
activity of fofolipase (PLA) was detected in fraction 14. This fraction was subjected to a
new phase chromatography on C18 analytical column, resulting in the isolation of a new
PLA. By electrophoresis, SDS-PAGE the enzyme showed high degree of purity, with
molecular mass around 13 kDa. The PLA2 activity showed optimal at pH of 8.2 and at
temperatures between 35-40 º C and remains stable up to temperatures of 60 º C.
According to our experiments isolated PLA was inhibited Ca2 +, and activity was not
changed by Cu2 +. Manaca extract, ellagic acid and quercetin strongly inhibited the PLA,
showing that this enzyme has potential for the selection of compounds with potential antiinflammatory.
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Citação
SANTANA, Pedro Henrique Camargo. Purification and characterization of a phospholipase A2 from the venom of Crotalus durissus yellow collilinetaus. 2009. 59 f. Dissertação (Mestrado em Ciências Biolóicas) - Universidade Federal de Goiás, Goiânia, 2009.