ABSTRACT
Protein C activator from Agkistrodon halys halys venom has been isolated and purified by ion-exchange and gel-filtration chromatography. The purified enzyme consists of the single peptide-chain with molecular weight of 34 kDa. Its pH-optimum was in 7.5-8.0 range. The enzyme was inhibited by DFP, benzamidine, PMSF, EGTA. Protein C activator was as effective as "Protac" (Pentapharm AG, Switzerland).
Subject(s)
Crotalid Venoms/chemistry , Peptides/isolation & purification , Protein C/analysis , Animals , Humans , Intercellular Signaling Peptides and Proteins , Partial Thromboplastin Time , Peptide Hydrolases/metabolism , Protease Inhibitors/pharmacology , Substrate Specificity , Thrombosis/blood , Thrombosis/prevention & controlABSTRACT
Snakes' venom is a mixture of biologically active substances, containing proteins and peptides. A number of these proteins interact with haemostasis system components. Activators and inhibitors affecting blood coagulation and fibrinolysis systems are of special interest. Venom components can be classified into three main groups, such as procoagulants, anticoagulants and fibrinolytic enzymes according to their action. This review is focused on enzymes from Agkistrodon halys halys venom. They are thrombine-like enzyme, named Ancystron-H, flbrinogenolytic enzyme, protein C activator and platelet aggregation inhibitor. Ancystron-H is used for determination of fibrinogen level in blood plasma of patients undergoing heparin treatment and blood coagulation inhibitors accumulation. The fibrinogenolytic enzyme can be used as the instrument for protein-protein interactions in fibrinogen-fibrin system. The protein C activator is used for protein C level determination in blood plasma with different pathologies. Functions of the platelet aggregation inhibitor, belonging to disintegrins group, can be used for development of antithrombotic preparations. Information about the use of snake venoms in science and medicine is presented.
Subject(s)
Enzymes/isolation & purification , Platelet Aggregation/drug effects , Snake Venoms/enzymology , Snakes/physiology , Ancrod/isolation & purification , Ancrod/pharmacology , Animals , Batroxobin/isolation & purification , Batroxobin/pharmacology , Enzymes/pharmacology , Humans , Protein C/analysis , Serine Endopeptidases/isolation & purification , Serine Endopeptidases/pharmacologyABSTRACT
Protein C activator from Agkistrodon halys halys venom has been purified by ion-exchange and gel-filtration chromatography. The purified enzyme consists of a single peptide chain with molecular weight of 34 kD. Its pH-optimum was the range of 7,5-8,0. The enzyme was inhibited by DFP, benzamidine, PMSF, EGTA. Protein C activator was as effective as Protac (Pentapharm AG, Switzerland) for determination of protein C level in blood plasma using APTT test and protein C chromogenic substrate.
Subject(s)
Crotalid Venoms/chemistry , Peptides/isolation & purification , Protein C/analysis , Animals , Chromatography, Gel , Disseminated Intravascular Coagulation/blood , Disseminated Intravascular Coagulation/diagnosis , Humans , Hydrogen-Ion Concentration , Intercellular Signaling Peptides and Proteins , Partial Thromboplastin Time , Peptides/antagonists & inhibitors , Peptides/metabolismABSTRACT
By Q-sepharose column ion-exchange chromatography, alkyl-sepharose column hydrophobic chromatography the purified fibrinogenolytic enzyme was obtained from Agkistrodon halys halys venom. It is a single peptide-chain with molecular weight about 28 kDa. It was founded that this enzyme cleaved A alpha-chain of fibrinogen, pH-optimum was determined in the range of 7.5-8.0. Its fibrinogenolytic activity was estimated 15.6 mM fibrinogen/min per mg protein; caseinolytic activity was estimated 7.5 c.u., and amidolytic activity was 0.325 mM pNA/min/mg and 0.175 mM pNA/min/mg for S2238 and S2251 respectively; K(m) was 5.6 mM. The enzyme activity was inhibited by DFP and benzamidine. These results suggest that the enzyme is serine protease. It inhibited the platelet-aggregation.
Subject(s)
Agkistrodon , Crotalid Venoms/enzymology , Fibrinolysis , Animals , Benzamidines/pharmacology , Caseins/metabolism , Crotalid Venoms/isolation & purification , Fibrin/metabolism , Fibrinogen/chemistry , Fibrinogen/metabolism , Hydrogen-Ion Concentration , Isoflurophate/pharmacology , Molecular Weight , Platelet Aggregation Inhibitors/pharmacology , Protease Inhibitors/pharmacology , Serine Proteinase Inhibitors/pharmacology , Substrate SpecificityABSTRACT
Fibrinogen is found to be able to produce highly molecular aggregates in solution. Its maximal quantity aggregates at the temperature of 4 degrees C. The pH dependence of fibrinogen aggregates production is shown. Highly molecular aggregates of fibrinogen are found to be produced mainly due to hydrogen bonds. Electron-microphotography of aggregated fibrinogen is presented. Complementary molecular loci responsible for the formation of ordered aggregates are discussed from the standpoint of their significance in the fibrin polymerization.
