Purification and Biochemical Characterization of Three Myotoxins fromBothrops mattogrossensisSnake Venom with Toxicity againstLeishmaniaand Tumor Cells

Abstract

Bothrops mattogrossensis snake is widely distributed throughout eastern South America and is responsible for snakebites in this region. This paper reports the purification and biochemical characterization of three new phospholipases A2 (PLA2s), one of which is presumably an enzymatically active Asp49 and two are very likely enzymatically inactive Lys49 PLA2 homologues. The purification was obtained after two chromatographic steps on ion exchange and reverse phase column. The 2D SDS-PAGE analysis revealed that the proteins have pI values around 10, are each made of a single chain, and have molecular masses near 13 kDa, which was confirmed by MALDI-TOF mass spectrometry. The N-terminal similarity analysis of the sequences showed that the proteins are highly homologous with other Lys49 and Asp49 PLA2s from Bothrops species. The PLA2s isolated were named BmatTX-I (Lys49 PLA2-like), BmatTX-II (Lys49 PLA2-like), and BmatTX-III (Asp49 PLA2). The PLA2s induced cytokine release from mouse neutrophils and showed cytotoxicity towards JURKAT (leukemia T) and SK-BR-3 (breast adenocarcinoma) cell lines and promastigote forms of Leishmania amazonensis. The structural and functional elucidation of snake venoms components may contribute to a better understanding of the mechanism of action of these proteins during envenomation and their potential pharmacological and therapeutic applications.

Bothrops jararaca and Bothrops erythromelas Snake Venoms Promote Cell Cycle Arrest and Induce Apoptosis via the Mitochondrial Depolarization of Cervical Cancer Cells

“…Other studies with adenocarcinoma cell lines of human colon LS174 (p53wt), HCT116 (p53wt), and HT29 (p53mut) found that Lebein (disintegrin heterodimeric) isolated from the venom of Macrovipera lebetina snake significantly inhibited cell viability [28]. Like the venoms of snakes of the genus Bothrops , the species B. mattogrossensis had three isolates of phospholipase A2 (PLA2) with cytotoxic activity in Jurkat lines (leukemia T) and SK-BR-3 (breast adenocarcinoma) [29]. But it is noteworthy that so far there are rare works in the literature exploring cervical carcinoma cell lines SiHa and HeLa, challenged with the venoms of B. jararaca and B. erythromelas snakes.…”

BmajPLA 2 -II, a basic Lys49-phospholipase A 2 homologue from Bothrops marajoensis snake venom with parasiticidal potential

“…Adade et al [26] demonstrated that the use of parabromophenacyl bromide (p-BPB), a PLA 2 inhibitor, caused partial inhibition of the trypanocidal activity of C. v. viridis venom. In three studies, PLA 2 s were isolated from B. brazili, B. mattagrossensis and B. moojeni venoms and their antiparasitic effects were checked against several different species of Leishmania [27][28][29]. Additionally, a non-catalytic PLA 2 isolated from Bothrops pauloensis venom, BnSP-7 inhibited the proliferation of L. amazonensis promastigotes and amastigotes and also caused morphological changes in the parasites [30].…”

l-amino acid oxidase from Bothrops marajoensis causes nephrotoxicity in isolated perfused kidney and cytotoxicity in MDCK renal cells

“…Bothrops venoms consist of a mixture of molecules of proteinaceous nature, including proteases, such as serine and metalloproteases, phospholipase A 2 (PLA2), 50-nucleotidase, hyaluronidase, C-type lectins and L-amino acid oxidase (LAAO) (Queiroz et al, 2008;Campos et al, 2013;de Moura et al, 2014).…”