Unmasking Snake Venom ofBothrops leucurus: Purification and Pharmacological and Structural Characterization of New PL Bleu TX-III

Abstract

Bleu TX-III was isolated from Bothrops leucurus snake venom on one-step analytical chromatography reverse phase HPLC, was homogeneous on SDS-PAGE, and was confirmed by Q-Tof Ultima API ESI/MS (TOF MS mode) mass spectrometry in 14243.8 Da. Multiple alignments of Bleu TX-III show high degree of homology with basic PLA2 myotoxins from other Bothrops venoms. Our studies on local and systemic myotoxicity “in vivo” reveal that Bleu TX-III is myotoxin with local but not systemic action due to the decrease in the plasmatic CK levels when Bleu TX-III is administrated by intravenous route in mice (dose 1 and 5 μg). And at a dose of 20 μg myotoxin behaves like a local and systemic action. Bleu TX-III induced moderate marked paw edema, evidencing the local increase in vascular permeability. The inflammatory events induced in the mice (I.M.) were investigated. The increase in the levels of IL-1, IL-6, and TNF-α was observed in the plasma. It is concluded that Bleu TX-III induces inflammatory events in this model. The enzymatic phospholipid hydrolysis may be relevant to these phenomena. Bothrops leucurus venom is still not extensively explored, and the knowledge of its toxins separately through the study of structure/function will contribute for a better understanding of its action mechanism.

PhTX-II a Basic Myotoxic Phospholipase A2 from Porthidium hyoprora Snake Venom, Pharmacological Characterization and Amino Acid Sequence by Mass Spectrometry

“…The PhTX-II PLA 2 is an enzyme composed of a unique polypeptidic chain revealed by Tricine SDS-PAGE with molecular mass of 14,149.08 Da confirmed by mass spectrometry ( Figure 2 ). This value is similar to other isolated myotoxic PLA 2 from snake venom [ 20 , 21 , 22 ].…”

“…These observations probably are due to that exons in PLA 2 genes mutate faster than introns, and this accelerated evolution of exons plays a significant role in the emergence of new isoenzymes by altering the target specificity. Similar results were found with Bothrops leucurus and Bothrops asper [ 9 , 20 , 24 ].…”

Cytotoxic and inflammatory potential of a phospholipase A2 from Bothrops jararaca snake venom

“…On the other hand, levels of LTB 4 , IL-10, TNF-α and nitric oxide were not altered after stimulation with BJ-PLA 2 -I. Taking all these findings into account, the results for BJ-PLA 2 -I indicate a local inflammatory response, similar to the ones previously described for other Asp49 PLA 2 s from Bothrops venoms, such as BatroxPLA 2 ( B. atrox ) [ 24 ], MT-III ( B. asper ) [ 6 ]; Bl-PLA 2 ( B. leucurus ) [ 30 ] and Bleu-TX-III ( B. leucurus ) [ 49 ].…”

Snake Venom Hemotoxic Enzymes: Biochemical Comparison between Crotalus Species from Central Mexico

“…Fractionation of venoms from the three species by RP–HPLC showed similar elution profiles, however, the concentrations among them were different. Abundant peaks ( ~10) between 40 and 55 min were found, where eluting times for proteins such as phospholipases A 2 (35–45 min) [57], metalloproteases and hyaluronidases (≥50 min) [58], have been reported. Peaks appearing at 20 and 30 min were less abundant (~6), those peaks have similar eluting times to the previously reported for some serine proteases [59].…”