Expression, purification and characterization of recombinant Jerdonitin, a P-II class snake venom metalloproteinase comprising metalloproteinase and disintegrin domains

Toxicon. 2010 Feb-Mar;55(2-3):375-80. doi: 10.1016/j.toxicon.2009.08.016. Epub 2009 Sep 2.

Abstract

Jerdonitin is a P-II class snake venom metalloproteinase comprising metalloproteinase and disintegrin domains. In this study, we established a high-level expression system in Pichia pastoris and developed a purification strategy for the recombinant Jerdonitin. This recombinant Jerdonitin degraded fibrinogen at a level of activity comparable with its wild type. The effects of recombinant Jerdonitin on inhibiting ADP-induced human platelet aggregation were in a dose-dependent manner with an IC(50) of 248nM. In addition, we reported here that Jerdonitin can significantly inhibit the growth of several cell lines, including human liver cancer cells (Bel7402), human leukemia cells (K562) and human gastric carcinoma cells (BGC823). This study offers recombinant Jerdonitin that will be valuable for further functional and structural studies of Jerdonitin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Coloring Agents
  • DNA, Complementary / biosynthesis
  • DNA, Complementary / genetics
  • Disintegrins / biosynthesis*
  • Disintegrins / chemistry
  • Disintegrins / pharmacology*
  • Dose-Response Relationship, Drug
  • Fibrinolytic Agents / pharmacology
  • Humans
  • Metalloproteases / biosynthesis*
  • Metalloproteases / chemistry
  • Metalloproteases / pharmacology*
  • Pichia / metabolism
  • Plasmids
  • Platelet Aggregation / drug effects
  • Protein Processing, Post-Translational
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / pharmacology
  • Snake Venoms / chemistry
  • Snake Venoms / enzymology*
  • Tetrazolium Salts
  • Thiazoles

Substances

  • Coloring Agents
  • DNA, Complementary
  • Disintegrins
  • Fibrinolytic Agents
  • Recombinant Proteins
  • Snake Venoms
  • Tetrazolium Salts
  • Thiazoles
  • Metalloproteases
  • thiazolyl blue