title: Simultaneous isolation of protein C activator, fibrin clot promoting enzyme (fibrozyme) and phospholipase A2 from the venom of the southern copperhead snake.
abstract: The simultaneous isolation of three enzymes from the southern copperhead snake venom (Agkistrodon contortrix contortrix; ACC) is described. The first step is a chromatography of crude venom on a Mono S cation-exchange column at pH 6.5. A fibrin clot promoting enzyme (fiprozyme) that preferentially releases fibrinopeptide B from fibrinogen is isolated from the fraction not binding to the Mono S by a further three-step process. The procedure involves affinity chromatography on Blue Sepharose, gel chromatography on Sephacryl S-200 and metal-chelate chromatography on Chelating Sepharose. Protein C activator and phospholipase coelute from the Mono S column. They are separated by a gel chromatography on Sephacryl S-200. After this step two enzymes are obtained: a highly purified protein C activator applicable in methods for determination of functional level of protein C (a plasma regulator of hemostasis) and an electrophoretically pure enzyme with the activity of phospholipase A2.
authors.affiliation: Department of Biochemistry, Institute of Hematology and Blood Transfusion, Prague, Czech Republic.
authors.forename: H
authors.initials: H
authors.lastname: Fortová
authors.forename: J
authors.initials: J
authors.lastname: Suttnar
authors.forename: J E
authors.initials: JE
authors.lastname: Dyr
authors.forename: J
authors.initials: J
authors.lastname: Pristach
mesh_terms: D017836:Agkistrodon; D000818:Animals; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003435:Crotalid Venoms / Q000737:chemistry*; D010450:Endopeptidases / Q000302:isolation & purification*; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides / Q000302:isolation & purification*; D010741:Phospholipases A / Q000302:isolation & purification*; D054467:Phospholipases A2
chemical_list: D003435:Crotalid Venoms; D036341:Intercellular Signaling Peptides and Proteins; D010455:Peptides; C049647:snake venom protein C activator; D010741:Phospholipases A; D054467:Phospholipases A2; D010450:Endopeptidases; C107415:fiprozyme