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The characterization of trans-pecos copperhead (Agkistrodon contortrix pictigaster) venom and isolation of two new dimeric disintegrins

Biologicals. 2016 Jul;44(4):191-197. doi: 10.1016/j.biologicals.2016.02.005. Epub 2016 Apr 8.

Abstract

The vast amounts of toxins within the venom of snakes, while known to cause medical emergencies, display various biological functions. Trans-pecos copperhead (Agkistrodon contortrix pictigaster) crude venom separated by cation-exchange chromatography showed several fractions with fibrinolytic, hemorrhagic, gelatinase and platelet activities. Venom fractions 1, 2, 4, 5, and 12-17 contained fibrinolytic activity. Venom fractions 1, 2, 5 and 12-14 had hemorrhagic activity. Fractions 1, 2, 12, 13 and 17 contained gelatinase activity. Reverse-Phase C18 High Performance Liquid Chromatography was also used to purify and isolated disintegrins from this venom. Anti-platelet aggregation activity of the C18 fractions collected and performed on whole human blood showed that they inhibited platelet aggregation in presence of several agonists. Results from both SDS-PAGE and N-terminal sequencing determined that pictistatin 1 obtained from the Trans-Pecos copperhead venom was a dimeric disintegrin, and pictistatin 2 was a heterodimeric disintegrin. The molecules with anti-platelet activity could be considered in the development of more effective drugs, for numerous blood-related diseases such as stroke, heart attacks, thrombosis, and other medical conditions. In this study, we are presenting the first report of the purification, isolation, and partial characterization of two new dimeric disintegrins isolated from the venom of trans-pecos copperhead.

Keywords: Aggregation; Dimeric disintegrins; Pictistatin; Platelets; Venom.

MeSH terms

  • Agkistrodon / metabolism*
  • Amino Acid Sequence
  • Animals
  • Cations
  • Cell Line, Tumor
  • Cell Proliferation / drug effects
  • Chromatography, High Pressure Liquid
  • Chromatography, Ion Exchange
  • Crotalid Venoms / chemistry
  • Crotalid Venoms / metabolism*
  • Crotalid Venoms / pharmacology
  • Disintegrins / chemistry
  • Disintegrins / isolation & purification*
  • Disintegrins / metabolism*
  • Electrophoresis, Polyacrylamide Gel
  • Fibrinolytic Agents / metabolism
  • Fibrinolytic Agents / pharmacology
  • Gelatinases / metabolism
  • Humans
  • Platelet Aggregation Inhibitors / metabolism
  • Platelet Aggregation Inhibitors / pharmacology
  • Protein Multimerization
  • Rabbits
  • Skin / blood supply
  • Skin / drug effects

Substances

  • Cations
  • Crotalid Venoms
  • Disintegrins
  • Fibrinolytic Agents
  • Platelet Aggregation Inhibitors
  • Gelatinases