Elucidation of procoagulant mechanism and pathophysiological significance of a new prothrombin activating metalloprotease purified from Daboia russelii russelii venom

Toxicon. 2015 Jun 15;100:1-12. doi: 10.1016/j.toxicon.2015.03.019. Epub 2015 Mar 27.

Abstract

The procoagulant proteases present in Russell's Viper venom (RVV) are responsible for promoting consumption coagulopathy in victims. In this study, a procoagulant metalloprotease (Rusviprotease) possessing prothrombin activating and α-fibrinogenase properties has been purified and characterized from RVV. Rusviprotease is a 26.8 kDa glycoprotein which also exists in other multimeric forms. The peptide mass fingerprinting and secondary structure analyses of Rusviprotease revealed its similarity with snake venom prothrombin activators and metalloproteases. Similar to group A prothrombin activators, Rusviprotease cleaved prothrombin independent of any co-factor requirement generating meizothrombin which is further cleaved to form thrombin. The Km and Vmax values of Rusviprotease towards prothrombin were determined to be 1.73 μM, and 153.5 nM thrombin generated/min/μmoles of Rusviprotease, respectively. The Km and Vmax values of Rusviprotease towards fibrinogen were calculated to be 3.14 μM and 78.7 nmol/min, respectively. Spectrofluorometric study provided the evidence of interaction between Rusviprotease and factor Xa with a Kd value of 6.64 nM. This interaction augmented the prothrombin activating property of the factor Xa-prothrombinase-Rusviprotease complex by 2.5 fold. Intravenous injection of Rusviprotease to BALB/c mice (0.1 mg/kg) resulted in in vivo defibrinogenation rendering the blood incoagulable. In conclusion, Rusviprotease is the first example of a prothrombin activator with fibrinogenolytic property purified from Daboia russelii russelii venom.

Keywords: Metalloprotease; Procoagulant protein; Prothrombin activator; Russell's Viper; Snake venom.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Coagulation / drug effects
  • Blood Platelets / drug effects
  • Chemical Fractionation
  • Coagulants / chemistry
  • Coagulants / isolation & purification
  • Coagulants / pharmacology*
  • Coagulants / toxicity
  • Erythrocytes / drug effects
  • Goats / blood
  • Metalloproteases / chemistry
  • Metalloproteases / isolation & purification
  • Metalloproteases / pharmacology*
  • Metalloproteases / toxicity
  • Mice, Inbred BALB C
  • Plasma / drug effects
  • Russell's Viper*
  • Toxicity Tests, Acute
  • Viper Venoms / chemistry*
  • Viper Venoms / toxicity

Substances

  • Coagulants
  • Viper Venoms
  • Metalloproteases