Isolation, Biochemical Characterization and Antiparasitic Activity of BmatTX-IV, A Basic Lys49-Phospholipase A2 from the Venom of Bothrops mattogrossensis from Paraguay

Abstract

Background: Functional and structural diversity of proteins of snake venoms is coupled with a wide repertoire of pharmacological effects. Snake venoms are targets of studies linked to searching molecules with biotechnological potential. Methods: A homologue phospholipase A2 (BmatTX-IV) was obtained using two chromatographic techniques. Mass spectrometry and two-dimensional gel electrophoresis were used to determine the molecular mass and isoelectric point, respectively. By means of Edman degradation chemistry, it was possible to obtain the partial sequence of amino acids that comprise the isolated toxin. Trypanocidal, leishmanicidal and cytoxic activity against Trypanosoma cruzi, Leishmania infantum and murine fibrobasts was determinated. Results: Combination of both chromatographic steps used in this study demonstrated efficacy to obtain the PLA2-Lys49. BmatTX-IV showed molecular mass and isoelectric point of 13.55 kDa and 9.3, respectively. Amino acid sequence of N-terminal region (51 residues) shows the presence of Lys49 residue at position 49, a distinctive trait of enzymatically inactive PLA2. Bothrops mattogrossensis snake venom showed IC50 values of 11.9 μg/mL against Leishmania infantum promastigotes and of 13.8 μg/mL against Trypanosoma cruzi epimastigotes, respectively. On the other hand, the venom showed a high cytotoxic activity (IC50 value of 16.7 μg/mL) against murine fibroblasts, whereas the BmatTX-IV showed IC50 value of 81.2 μg/mL. Conclusion: Physicochemical and biological characterization of snake venoms components is critically important, since these complex mixtures provide a source of molecules with antiparasitic potential, making further studies necessary to identify and characterize components with higher efficacy and selectivity.

Antimalarial activity of basic phospholipases A2 isolated from Paraguayan Bothrops diporus venom against Plasmodium falciparum

“…2 A–C). The combination of IEC followed by RPC has already been reported by several authors, who mention that these steps contribute considerably to the isolation of basic PLA 2 s from bothropic venoms, guaranteeing the purification of the molecules of interest with a high degree of purity ( Alfonso et al, 2019 ; Grabner et al, 2017 ; Nunes et al, 2013 ). The electrophoretic profile of the isolated toxins can be seen in Fig.…”

“…As previously described, svPLA 2 s present in B. diporus venom are responsible for different pharmacological effects. While the literature reports the antiparasitic activity of this group of proteins ( Alfonso et al, 2019 ; Grabner et al, 2017 ; Nunes et al, 2013 ; Stábeli et al, 2006 ), this study aimed to evaluate the cytotoxic capacity of PLA 2 s against the intra-erythrocytic forms of P. falciparum , becoming the first report in scientific literature to perform antiplasmodial assays of toxins isolated from Paraguayan B. diporus .…”

Suicidal Leishmania

“…In cytotoxicity assays, its activity has also been demonstrated against several Leishmania spp. [41][42][43]. The pilot TMP/TMP-lac treatment experiments with the BnSP-7-ecDHFR-HA-expressing parasites have confirmed the anti-leishmanial potential of this toxin (Figure 1).…”

Antiprotozoal Effect of Snake Venoms and Their Fractions: A Systematic Review

“…Previous reports [34,47,71] indicated the inhibitory effects of PLA2s of the respective snake venoms on P. falciparum. According to many other studies [21,58,59,68], various PLA2s inhibited the cellular viability of Leishmania species. In addition, Borges et al [29] and Borges et al [30] reported that PLA2s of B. pauloensis inhibited parasite adhesion, intracellular proliferation, parasite viability, intracellular proliferation and proinflammatory cytokine production in T. gondii.…”

“…We identified 55 studies that met our inclusion criteria, of which 20 were conducted to evaluate the antiprotozoal efficacy of snake crude venoms [18,21,28,[34][35][36]38,40,43,[45][46][47]49,50,58,[61][62][63]65,69]. We identified a previous study that reported the antiprotozoal effect of snake crude venoms, with some hypotheses about the specific snake venom fractions responsible for antiprotozoal activity without proof from laboratory trials [10].…”