BaltPLA2: A New Phospholipase A2 from Bothrops alternatus Snake Venom with Antiplatelet Aggregation Activity

Background: In last decades, snake venoms have aroused great interest of the medicine due to the pathophysiological effects caused by their toxins. These include the phospholipases A(2), low molecular weight proteins capable of causing haemorrhagic, myotoxic, inflammatory and neurotoxic effects after an ophidian accident. The present work describes the isolation and biochemical characterization of a new PLA(2) isolated from the B. alternatus snake venom, which was named BaltPLA(2). Method: The rapid and efficient purification of this toxin was performed using only two chromatography steps (anion exchange and hydrophobic chromatography). Results: BaltPLA(2) is an acidic protein (pI 4.4) with an apparent molecular mass of 17000 (SDS-PAGE) and 14074.74 Da (MALDI TOF/TOF). Analysis of fragments ion by MS/MS showed the following internal amino acid sequence SGVIICGEGTPCEK, which did not exhibit homology with other PLA(2) from the same venom. BaltPLA(2) is a catalytically active, which displayed an anticoagulant action, inhibition of platelet aggregation induced by epinephrine (similar to 80%) and ADP (24%). BaltPLA(2) also was able to induce myonecrosis and the release of cytokines (IL-10, IL-12 and TNF-alpha) in macrophages culture. Conclusion: The results presented in this work greatly contribute to a better understanding of the mechanism of enzymatic and pharmacological actions of PLA(2)s from snake venoms and they may contribute to its application in medical research.