Cytotoxic, thrombolytic and edematogenic activities of leucurolysin-a, a metalloproteinase from Bothrops leucurus snake venom

Toxicon. 2007 Jul;50(1):120-34. doi: 10.1016/j.toxicon.2007.03.002. Epub 2007 Mar 14.

Abstract

Leucurolysin-a (leuc-a), a 23 kDa non-hemorrhagic metalloproteinase, is found in venom of the viper Bothrops leucurus. Here, we examine the biological consequences of leuc-a, including thrombolytic activity, direct effects on endothelial cells in culture and edematogenic activity in vivo. We demonstrate fibrinolytic activity of leuc-a, in which the protease specifically degrades alpha, beta, and gamma-gamma chains. While not causing hemorrhaging, leuc-a does cause thrombolytic activities in whole blood clots. Endothelial cells are highly resistant to leuc-a in culture. Cell viability suffered only when cells were exposed to large quantities of the protease. Nevertheless, leuc-a induces changes in cell morphology. The impact of leuc-a on cell adhesion was confirmed by an adhesion assay, in which cell adhesion to fibronectin decreased due to leuc-a. This mild cellular impact is unlike that of crude venom, where lower concentrations triggered cell death and a greater reduction in cell adhesion. Also, leuc-a increased microvessel permeability with marked edema in mice peritoneum and foot pads. These effects are similar to those of other P-I class SVPMs. These in vivo effects were weaker when crude venom was tested. In conclusion, albeit not showing significant hemorrhagic activity, leuc-a can induce a prominent edema which appears to be significant in the local effects observed after B. leucurus venom accidents.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Analysis of Variance
  • Animals
  • Bothrops / metabolism*
  • Cell Adhesion / drug effects
  • Cell Survival
  • Cells, Cultured
  • Crotalid Venoms / enzymology
  • Crotalid Venoms / isolation & purification
  • Crotalid Venoms / metabolism
  • Crotalid Venoms / toxicity*
  • Edema
  • Fibrin / metabolism
  • Fibrinolysis / drug effects*
  • Fibronectins / metabolism
  • Flow Cytometry
  • Humans
  • Metalloproteases / isolation & purification
  • Metalloproteases / metabolism
  • Metalloproteases / toxicity*
  • Microvessels / metabolism
  • Rabbits
  • Thrombin / metabolism

Substances

  • Crotalid Venoms
  • Fibronectins
  • Fibrin
  • Metalloproteases
  • Thrombin