BbMP-1, a new metalloproteinase isolated from Bothrops brazili snake venom with in vitro antiplasmodial properties

Abstract

This study describes the biochemical and functional characterization of a new metalloproteinase named BbMP-1, isolated from Bothrops brazili venom. BbMP-1 was homogeneous on SDS-PAGE, presented molecular mass of 22,933Da and pI 6.4. The primary structure was partially elucidated with high identity with others metalloproteinases from Viperidae venoms. The enzymatic activity on azocasein was evaluated in different experimental conditions (pH, temperature). A significant reduction in enzyme activity after exposure to chelators of divalent cations (EDTA), reducing agents (DTT), pH less than 5.0 or temperatures higher than 45 °C was observed. BbMP-1 showed activity on fibrinogen degrading Aα chain quickly and to a lesser extent the Bβ chain. Also demostrated to be weakly hemorrhagic, presenting however, significant myotoxic and edematogenic activity. The in vitro activity of BbMP-1 against Plasmodium falciparum showed an IC50 of 3.2 ± 2.0 μg/mL. This study may help to understand the pathophysiological effects induced by this group of toxin and their participation in the symptoms observed in cases of snake envenomation. Moreover, this result is representative for this group of proteins and shows the biotechnological potential of BbMP-1 by the demonstration of its antiplasmodial activity.

Venomics and antivenomics of the poorly studied Brazil’s lancehead, Bothrops brazili (Hoge, 1954), from the Brazilian State of Pará

“…In the murine model, Peruvian B. brazili exhibited minimum hemorrhagic dose (MHD) of 7.40 μg/ mouse), minimum dermonecrotic dose (MND) of 152.15 μg/ mouse, minimum coagulant dose against plasma (MCD-P) and fibrinogen (MCD-F) of 19.20 and 1020.0 μg/mL, respectively, and minimum defibrinogenating dose (MDD) of 7.0 μg/mouse [11]. Although described as a new Bothrops from Brazil 65 years ago [15], very few studies have been reported on the toxin arsenal of the Brazil's lancehead venom, and these were mainly focused on the pharmacological effects and possible biotechnological applications of isolated toxins [21][22][23][24][25][26][27][28][29][30][31], including acidic and basic phospholipase A 2 (PLA 2 ) molecules (myotoxic Braziliase I and II, MTX I and II, brazilitoxins II and III) [23][24][25][26]; a PI-snake venom metaloproteinase (SVMP), with in vitro antiplasmodial properties [27]; coagulant thrombin-like and pro-angiogenic snake venom serine proteinase (SVSP) [28,29]; and a hyaluronidase [30].…”

In vitro trichomonocidal potency of Naja nigricollis and Bitis arietans snake venom

“…A similar study on BMP-1, a new metalloproteinase isolated from Bothrops Brazili snake venom has an in vitro anti-plasmodial property against P. falciparum with an IC 50 of 3.2±2.0 mg/mL [29]. Recent literature described L-MAO isolated from Bothrops pirajai and Bothrops alternatus venoms had inhibited Escherichia coli growth.…”

“…Recent literature described L-MAO isolated from Bothrops pirajai and Bothrops alternatus venoms had inhibited Escherichia coli growth. Interestingly purified L-MAO of Australian elapid, Pseudechis australis (Australian king brown or mulga snake) also had antibacterial property [29]. Studies in this context include the effect of Crotalus viridis viridis snake venom on Trypanosoma cruzi, which shows a promising result of 76-93% reduction in the number of parasites per infected cell and a 94-97.4% reduction in the number of parasites per 100 cells after 96 h of infection [30].…”

Functional Elucidation of Nemopilema nomurai and Cyanea nozakii Nematocyst Venoms’ Lytic Activity Using Mass Spectrometry and Zymography

“…The effects of metalloproteinase inhibitors, EDTA and batimastat, on the metalloproteinase activity of jellyfish venom were measured with azocasein [ 74 ]. Briefly, 100 μL of the reaction mixture contained 10 μL of EDTA (final concentration, 31.2 μM–10000 μM) or batimastat (final concentration, 6.5 μM–209.4 μM), 10 μL of NnNV (3.16 μg/μL) and 80 μL of azocasein (5 mg/mL) resuspended in assay buffer (50 mM Tris, 100 mM NaCl, 5 mM CaCl 2 , pH 8.8).…”