Bbil-TX, a PLA2, was purified fromBothriopsis bilineatasnake venom after only one chromatographic step using RP-HPLC onμ-Bondapak C-18 column. A molecular mass of 14243.8 Da was confirmed by Q-Tof Ultima API ESI/MS (TOF MS mode) mass spectrometry. The partial protein sequence obtained was then submitted to BLASTp, with the search restricted to PLA2from snakes and shows high identity values when compared to other PLA2s. PLA2activity was presented in the presence of a synthetic substrate and showed a minimum sigmoidal behavior, reaching its maximal activity at pH 8.0 and 25–37∘C. Maximum PLA2activity required Ca2+and in the presence of Cd2+, Zn2+, Mn2+, and Mg2+it was reduced in the presence or absence of Ca2+. Crotapotin fromCrotalus durissus cascavellarattlesnake venom and antihemorrhagic factor DA2-II fromDidelphis albiventrisopossum sera under optimal conditions significantly inhibit the enzymatic activity. Bbil-TX induces myonecrosis in mice. The fraction does not show a significant cytotoxic activity in myotubes and myoblasts (C2C12). The inflammatory events induced in the serum of mice by Bbil-TX isolated fromBothriopsis bilineatasnake venom were investigated. An increase in vascular permeability and in the levels of TNF-a, IL-6, and IL-1 was was induced. Since Bbil-TX exerts a stronger proinflammatory effect, the phospholipid hydrolysis may be relevant for these phenomena.
Biochemical, Pharmacological, and Structural Characterization of New Basic Bbil-TX fromBothriopsis bilineataSnake Venom
