Poster 82.  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.441.

 

 

Enzymatic and Pharmacological Characterization of two Acidic Atoxic Phospholipase A2 Isolated from Bothrops moojeni Snake Venom.

 

1Silveira, L. B.; 1Fernandes, V. C.; 1Coppede, J. S.; 3Sant`Ana, C. D.; 1,2Marcussi, S.; 1Silva, E. G.; 1Araújo, A. L.; 1França, S. C.; 2Giglio, J. R. and 1Soares, A. M.

 

1 Unidade de Biotecnologia-UNAERP, Ribeirão Preto-SP; 2 Depto Bioquímica e Imunologia, FMRP-USP; 3 Depto. Análises Clínicas, Toxicológicas e Bromatológicas-FCFRP-USP, Ribeirão Preto-SP, Brasil.

 

Phospholipases A2 (PLA2s) are enzymes that catalyse the hydrolysis of 2-acyl-ester bonds of phospholipids. They are widely distributed in mammalian tissues, microorganisms, as well as reptile and arthropod venoms, having been extensively studied because of their relative abundance, in addition to their enzymatic and diversified biological activities. This work reported the isolation and enzymatic/pharmacological characterization of two acidic PLA2 isoforms (Bmoo-I-PLA2 and Bmoo-II-PLA2) from B. moojeni snake venom. The enzymes were purified by ion-exchange chromatography on CM-Sepharose, followed by rechromatography of the active fraction on Phenyl-Sepharose. The enzymatic active protein was assayed for purity by PAGE and RP-HPLC. PLA2 activity was measured an agar gel supplemented with egg yolk and human erythrocytes, under different conditions of temperature, pH, and presence of several divalent cations and inhibitors. In addition, the edema-inducing, myotoxic and coagulant activities of both enzymes were measured. They showed to be atoxic, although inducing edema in the mouse paw in a time-dose dependent mode and showing moderate (Bmoo-I-PLA­2) to high (Bmoo-II-PLA2) anticoagulant activity in poor platelet plasma, both enzymes induced collagen and ADP-dependent platelet aggregation inhibition. Snake venom PLA2s are multifunctional enzymes representing a high biotechnical potential and a molecular model for the study of intra and extra cellular action mechanism to ward their target tissues or molecules.

 

Financial support: FAPESP, CAPES, CNPq and UNAERP.

 

Correspondence to: luckinhas@netsite.com.br or andreimar@unaerp.br