Poster 14.  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.373.

 

 

Preliminary kinetic studies of a new isolated PLA2 basic from Lachesis muta muta

 

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

 

Department of Biochemistry, institute of Biology, State University of Campinas (UNICAMP), Campinas, SP, Brazil. 

 

The main objective of this work is the purification and partial enzymatic characterization of PLA2 from Lachesis muta muta.

PLA2 was isolated from Lachesis muta muta venom by combination of LC molecular exclusion chromatography and RP-HPLC. Amino acid analyses revealed a high contend of basic and hydrophobic amino acids (Lys, Gly and Tyr). SDS PAGE showed that PLA2 has an apparent molecular mass of aproximatelly 15 kDa. The measurement of enzymatic activity was made according to described by Holzer and Mackesy (1996) modified by Ponce-Soto, L.A. (2002). The enzymatic assays were made on VersaMAX 190 (Molecular Devices, CA. USA). The specific activity was 4,08 ± 0,09 nmoles/min/mg. The optimum pH was 8.0 and optimum temperature was 38șC to enzyme activity. The optimum incubation time of reaction was 20min. This PLA2 shows high enzymatic activity in the presence of Mn+2 and Mg+2 but was low in the presence of Zn+2 and Cu+2.

The enzymatic activity of PLA2 isolated from Lachesis muta muta has similar enzymatic properties of other PLA2 isolated from snake venom.

Support: CAPES, FAPESP and CNPq.

Correspondence to: dcdamico@unicamp.br