Bp-13 [PLA.sub.2]: purification and neuromuscular activity of a new Asp49 toxin isolated from Bothrops pauloensis snake venom

A new [PLA.sub.2] (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on µ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of...

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Published in:Biochemistry research international 2015-01
Main Authors: Sucasaca-Monzon, Georgina, Randazzo-Moura, Priscila, Rocha, Thalita, Torres-Huaco, Frank Denis, Vilca-Quispe, Augusto, Ponce-Soto, Luis Alberto, Marangoni, Sergio, da Cruz-Hofling, Maria Alice, Rodrigues-Simioni, Lea
Format: Article
Language:English
Subjects:
Health aspects
Lactic acid
Toxins
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Summary:A new [PLA.sub.2] (Bp-13) was purified from Bothrops pauloensis snake venom after a single chromatographic step of RP-HPLC on µ-Bondapak C-18. Amino acid analysis showed a high content of hydrophobic and basic amino acids and 14 half-cysteine residues. The N-terminal sequence showed a high degree of homology with basic Asp49 [PLA.sub.2] myotoxins from other Bothrops venoms. Bp-13 showed allosteric enzymatic behavior and maximal activity at pH 8.1, 36°-45°C. Full Bp-13 [PLA.sub.2] activity required [Ca.sup.2+]; its [PLA.sub.2] activity was inhibited by [Mg.sup.2+], [Mn.sup.2+], [Sr.sup.2+], and [Cd.sup.2+] in the presence and absence of 1mM [Ca.sup.2+]. In the mouse phrenic nervediaphragm (PND) preparation, the time for 50% paralysis was concentration-dependent (P < 0.05). Both the replacement of [Ca.sup.2+] by [Sr.sup.2+] and temperature lowering (24°C) inhibited the Bp-13 [PLA.sub.2]-induced twitch-tension blockade. Bp-13 [PLA.sub.2] inhibited the contractile response to direct electrical stimulation in curarized mouse PND preparation corroborating its contracture effect. In biventer cervicis preparations, Bp-13 induced irreversible twitch-tension blockade and the KCl evoked contracture was partially, but significantly, inhibited (P > 0.05). The main effect of this new Asp49 [PLA.sub.2] of Bothrops pauloensis venom is on muscle fiber sarcolemma, with avian preparation being less responsive than rodent preparation. The study enhances biochemical and pharmacological characterization of B. pauloensis venom.
ISSN:2090-2247
DOI:10.1155/2015/826059