Abstract: In this work, we describe the molecular cloning and pharmacological properties of an acidic phospholipase A2 (PLA2) isolated from Bothrops pauloensis snake venom. This enzyme, denominated BpPLA2-TXI, was purified by four chromatographic steps and represents 2.4% of the total snake venom protein content. BpPLA2-TXI is a monomeric protein with a molecular mass of 13.6 kDa, as demonstrated by Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) analysis and its theoretical isoelectric point was 4.98. BpPLA2-TXI was catalytically active and showed some pharmacological effects such as inhibition of platelet aggregation induced by collagen or ADP and also induced edema and myotoxicity. BpPLA2-TXI displayed low cytotoxicity on TG-180 (CCRF S 180 II) and Ovarian Carcinoma (OVCAR-3), whereas no cytotoxicity was found in regard to MEF (Mouse Embryonic Fibroblast) and Sarcoma 180 (TIB-66). The N-terminal sequence of forty-eight amino acid residues was determined by Edman degradation. In addition, the complete primary structure of 122 amino acids was deduced by cDNA from the total RNA of the venom gland using specific primers, and it was significantly similar to other acidic D49 PLA2s. The phylogenetic analyses showed that BpPLA2-TXI forms a group with other acidic D49 PLA2s from the gender Bothrops, which are characterized by a catalytic activity associated with anti-platelet effects.
Keywords: Bothrops pauloensis; phospholipase A2; molecular cloning
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Ferreira, F.B.; Gomes, M.S.R.; de Souza, D.L.N.; Gimenes, S.N.C.; Castanheira, L.E.; Borges, M.H.; Rodrigues, R.S.; Yoneyama, K.A.G.; Brandeburgo, M.I.H.; Rodrigues, V.M. Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom. Toxins 2013, 5, 2403-2419.
Ferreira FB, Gomes MSR, de Souza DLN, Gimenes SNC, Castanheira LE, Borges MH, Rodrigues RS, Yoneyama KAG, Brandeburgo MIH, Rodrigues VM. Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom. Toxins. 2013; 5(12):2403-2419.
Ferreira, Francis B.; Gomes, Mário S.R.; de Souza, Dayane L.N.; Gimenes, Sarah N.C.; Castanheira, Letícia E.; Borges, Márcia H.; Rodrigues, Renata S.; Yoneyama, Kelly A.G.; Brandeburgo, Maria I.H.; Rodrigues, Veridiana M. 2013. "Molecular Cloning and Pharmacological Properties of an Acidic PLA2 from Bothrops pauloensis Snake Venom." Toxins 5, no. 12: 2403-2419.