, 1993) has an MHD of 0.2 μg, B-JussuMP-I from Bothrops jararacussu has an MHD of 4 μg ( Mazzi et al., 2006) and BaH4 from Bothrops asper has an MHD of 2 μg ( Franceschi et al., 2000). Based on these results, we consider Batroxase to be a weakly hemorrhagic metalloproteinase.

To determine the mechanism underlying the induction of hemorrhage Selleckchem R428 by Batroxase, its capacity to digest extracellular matrix components was assessed. Batroxase was able to hydrolyze type IV collagen and fibronectin molecules, and it also degraded the α 1, α and γ chains of laminin in Matrigel, but it was not able to digest isolated laminin. No nidogen proteolysis was detected. According to Bou-Gharios et al. (2004), the basement membranes of blood vessels consist mainly of laminin, collagen and fibronectin. Therefore, the ability of Batroxase to hydrolyze these components is consistent with its ability to induce hemorrhage by degrading extracellular matrix components of the blood vessel basement

membranes. Batroxase was able to digest fibrinogen by cleaving the α and β chains. Furthermore, the fibrinogen hydrolysis occurred in a concentration-dependent manner and was inhibited by EDTA and EGTA, which indicates that its metalloproteinase character was important for inducing proteolysis. According to Mosesson (2005), under physiological conditions, fibrin is formed by the cleavage of the fibrinogen α chain by thrombin. However, the results obtained showed that α and β chain cleavage by Batroxase suggests that the fibrin formed

might not be able to polymerize. Thus, the activity of Batroxase on the fibrinogen molecule likely indicates a consumption of this substrate PR-171 nmr and an inhibition of clot and thrombus formation. Several PI SVMPs are able to preferentially digest the α chain of the fibrinogen molecule, e.g., BnPI from Bothrops neuwiedi ( Baldo et al., 2008), BlaH1 from Bothrops lanceolatus ( Stroka et al., 2005), Atroxlysin-I from Bothrops atrox ( Sanchez et al., 2010), BmooMPα-I from Bothrops moojeni ( Bernardes et al., 2008) and Neuwiedase from Bothrops neuwiedi ( Rodrigues et al., 2001). Fibrinolytic activity has been reported for several PI-class SVMPs, such as Neuwiedase www.selleck.co.jp/products/Paclitaxel(Taxol).html (Rodrigues et al., 2001) and BnP1 from Bothrops neuwiedi ( Baldo et al., 2008), Bothrojaractivase from Bothrops jararaca ( Berguer et al., 2008), Berythractivase from Bothrops erythromelas ( Silva et al., 2003), BthMP from Bothrops moojeni ( Lopes et al., 2009) and Atroxlysin-1 from Bothrops asper ( Sanchez et al., 2010). Batroxase was able to induce fibrin digestion in a concentration-dependent manner up to 8 μg. The lack of further digestion at higher concentrations was probably the result of the total consumption of the fibrin in the gel. To confirm that the fibrinolytic hydrolysis mediated by Batroxase was not the result of the activation of plasminogen to generate plasmin, Batroxase was incubated with plasminogen, and the resulting fragments were analyzed.