Isolation and functional characterization of proinflammatory acidic phospholipase A2 from Bothrops leucurus snake venom

Comp Biochem Physiol C Toxicol Pharmacol. 2011 Sep;154(3):226-33. doi: 10.1016/j.cbpc.2011.06.003. Epub 2011 Jun 15.

Abstract

In the present study, an acidic PLA(2), designated Bl-PLA(2), was isolated from Bothrops leucurus snake venom through two chromatographic steps: ion-exchange on CM-Sepharose and hydrophobic chromatography on Phenyl-Sepharose. Bl-PLA(2) was homogeneous on SDS-PAGE and when submitted to 2D electrophoresis the molecular mass was 15,000Da and pI was 5.4. Its N-terminal sequence revealed a high homology with other Asp49 acidic PLA(2)s from snake venoms. Its specific activity was 159.9U/mg and the indirect hemolytic activity was also higher than that of the crude venom. Bl-PLA(2) induced low myotoxic and edema activities as compared to those of the crude venom. Moreover, the enzyme was able to induce increments in IL-12p40, TNF-α, IL-1β and IL-6 levels and no variation of IL-8 and IL-10 in human PBMC stimulated in vitro, suggesting that Bl-PLA(2) induces proinflammatory cytokine production by human mononuclear cells. Bothrops leucurus venom is still not extensively explored and knowledge of its components will contribute for a better understanding of its action mechanism.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Bothrops*
  • Cells, Cultured
  • Chromatography, Ion Exchange / methods
  • Crotalid Venoms / enzymology*
  • Cytokines / metabolism
  • Edema / chemically induced
  • Electrophoresis, Polyacrylamide Gel
  • Humans
  • Inflammation / metabolism*
  • Interleukin-10 / metabolism
  • Interleukin-12 Subunit p40 / metabolism
  • Interleukin-6 / metabolism
  • Interleukin-8 / metabolism
  • Leukocytes, Mononuclear / drug effects
  • Leukocytes, Mononuclear / metabolism
  • Male
  • Mice
  • Molecular Weight
  • Phospholipases A2 / chemistry*
  • Phospholipases A2 / isolation & purification
  • Phospholipases A2 / pharmacology*
  • Sepharose / analogs & derivatives
  • Sequence Homology, Amino Acid
  • Tumor Necrosis Factor-alpha / metabolism

Substances

  • Crotalid Venoms
  • Cytokines
  • Interleukin-12 Subunit p40
  • Interleukin-6
  • Interleukin-8
  • Tumor Necrosis Factor-alpha
  • Interleukin-10
  • phenyl-sepharose
  • Sepharose
  • Phospholipases A2