The PLA2 activity of L. muta rhombeata whole venom, Sephadex G75 fraction (FIII) and LmrTX was 122.70 ± 13.41; 191.65 ± 3.34 and 789.74 ± 6.59 nmol/mg/min, respectively, as shown in Fig. 4. The alkylation of histidine Microbiology inhibitor residues (with

p-bromophenacyl bromide) from LmrTX, reduced significantly the toxin PLA2 activity (only 11% of residual activity), 86.47 ± 13.41 nmol/mg/min (p < 0.05; Fig. 4). Snake venom phospholipases A2 (PLA2) are an extremely important and diverse group of proteins that affects hemostasis. In this study, we examined the ability of the LmrTX in altering the thrombus formation in living mouse. For this, we used a photochemically induced arterial thrombosis model in mice. Fig. 5 shows the effect of LmrTX in thrombus formation in the carotid artery of mice. Control animals that did not receive the protein injection showed a normal occlusion time, which was 57 ± 7.8 min. As shown in Fig. 5, LmrTX, the PLA2 from L. muta rhombeata venom, caused a change in the normal occlusion time. With doses of 7.5 and 15 μg/mice, the occlusion time was 99 ± 10 min and 94 ± 11.5 min respectively. The dose of 3.25 μg/mice did not significantly differ from control values (62.6 ± 10 min). The animals that were treated with the modified protein showed the occlusion time similar to control animals (64.4 ± 14.0 min). Anticoagulant PLA2s (mainly

Asp49 PLA2) have been described in all major snake groups. In in vitro condition, LmrTX showed anticoagulant activity (APTT), prolonging the normal clotting time of platelet selleckchem poor plasma ( Fig. 6A). When mice were pretreated with LmrTX at different times before determining APTT, the protein showed significant anticoagulant activity with a rapid onset PLEK2 (maximal response obtained at 15 min pos-injection) which was sustained during 1 h ( Fig. 6B). In the other hand, no significant effect on PT in vitro ( Fig. 6C) and ex vivo ( Fig. 6D) was observed. We also verified if LmrTX could interfere with platelet aggregation. The animals that received the protein (15 μg) showed a partial inhibition in ADP and thrombin-induced

washed platelet aggregation, 43 and 44%, respectively (Fig. 7). In snake venom PLA2 enzymes, His48 is conserved and plays a crucial role in the hydrolysis of phospholipids. Modification of PLA2 enzyme from L. muta rhombeata venom on His48 residue by alkylation leads to the 89% of reduction in enzymatic activity, with concomitant loss of anticoagulant effects in vitro. Accidents caused by Lachesis venom present many symptoms like local pain, oedema, haemorrhage and necrosis at the site of the bite. Moreover, systemic complications such as nausea, vomiting, diarrhea, hypotension and bradycardia, coagulation disturbances and renal failure are observed during Lachesis envenomation ( Jorge et al., 1997; Rucavado et al., 1999). Only a few proteins were purified from the venom of L.