Blood coagulation induced by the venom of Bothrops atrox. 2. Identification, purification, and properties of two factor X activators

Biochemistry. 1987 Feb 10;26(3):780-7. doi: 10.1021/bi00377a019.

Abstract

We have characterized and purified the two components of the venom of Bothrops atrox that activate the coagulation factor X. Activator 1 and activator 2 were separated by ion-exchange chromatography but otherwise presented similar characteristics. They consist of a heavy polypeptide of Mr 59,000 and either one or two light chains forming a doublet of Mr 14,000-15,000. They are inactive on synthetic substrates and on prothrombin or fibrinogen and thus appear to act specifically on factor X. They are not sensitive to inhibitors of serine proteases or thiol esterases. The activation of factor X is activated by Ca2+ ions with a Hill coefficient of 2.4 and is inhibited by Hg2+, Ba2+, and Cd2+. Its pH dependency suggests that the activity depends on the ionization of a group with an apparent pK of 6.9. We studied the cleavage of purified bovine factor X by B. atrox activators and compared it to that obtained with the factor X activator from Vipera russelli venom. Like the physiological activators, the venom's activators cleave the heavy chain of factor X, producing the activated factor Xa alpha. They produce however two other cleavages: one near the N-terminal end of the heavy chain of factor X, generating factor Xmu, and a second one located at one extremity of the heavy chain of factor Xa alpha, generating factor Xav.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blood Coagulation / drug effects*
  • Calcium / pharmacology
  • Cattle
  • Chromatography, DEAE-Cellulose
  • Chromatography, Gel
  • Crotalid Venoms / pharmacology*
  • Endopeptidases / isolation & purification
  • Endopeptidases / pharmacology*
  • Enzyme Activation
  • Factor X / drug effects
  • Factor X / metabolism*
  • Kinetics
  • Metalloendopeptidases*
  • Molecular Weight
  • Phospholipids / pharmacology

Substances

  • Crotalid Venoms
  • Phospholipids
  • Factor X
  • Endopeptidases
  • Metalloendopeptidases
  • factor X activator, Bothrops atrox venom
  • Calcium