Biochemical, Pharmacological, and Structural Characterization of New Basic P L A 2 Bbil-TX from Bothriopsis bilineata Snake Venom

Bbil-TX, a PLA 2 , was purified from Bothriopsis bilineata snake venom after only one chromatographic step using RP-HPLC on μ -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 PLA 2 from snakes and shows high identity values when compared to other PLA 2 s. PLA 2 activity 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 PLA 2 activity required Ca 2+ and in the presence of Cd 2+ , Zn 2+ , Mn 2+ , and Mg 2+ it was reduced in the presence or absence of Ca 2+ . Crotapotin from Crotalus durissus cascavella rattlesnake venom and antihemorrhagic factor DA2-II from Didelphis albiventris opossum 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 from Bothriopsis bilineata snake 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.