Platelet procoagulant properties studied with snake venom prothrombin activators.
Purified snake venom prothrombin activators were used to probe the procoagulant properties of platelet membranes. Human platelets were able to stimulate prothrombin activation by the venom activators from Oxyuranus scutellatus and Notechis scutatus, while the prothrombin activator from Echis carinatus was not affected by the presence of platelets. The prothrombin-converting activity of platelets was further studied with the venom activator from Oxyuranus scutellatus and with the factor Xa-Va complex as prothrombin activating enzymes. Stimulation of platelets with collagen, collagen plus thrombin or with the Ca-ionophore A23187 resulted in a considerable increase of platelet prothrombin converting activity probed with the factor Xa-Va complex as well as with the prothrombin activator from Oxyuranus scutellatus. The stimulatory effect of activated platelets on the rates of prothrombin activation by Oxyuranus scutellatus was similar to that determined for factor Xa-Va-catalyzed prothrombin activation. Compared to non-stimulated platelets, platelets stimulated with thrombin plus collagen exposed 20-times more procoagulant sites for as well the factor Xa-Va complex, as for the venom activator from Oxyuranus scutellatus. The actual number of procoagulant sites per platelet determined with the factor Xa-Va complex was in close agreement with the number of sites determined with the venom activator. Also the time course of appearance of procoagulant activity during platelet stimulation by collagen plus thrombin was comparable for both activator complexes. Phospholipase A2 treatment of stimulated platelets resulted in an almost complete loss of their ability to stimulate prothrombin activation by the enzyme from Oxyuranus scutellatus or by factor Xa-Va complex.(ABSTRACT TRUNCATED AT 250 WORDS)
Speijer
H
H
Department of Biochemistry, University of Limburg, Maastricht, The Netherlands.
Govers-Riemslag
J W
JW
Zwaal
R F
RF
Rosing
J
J
D001665:Binding Sites; D001779:Blood Coagulation Factors / Q000032:analysis / Q000378:metabolism*; D001792:Blood Platelets / Q000502:physiology / Q000648:ultrastructure; D002462:Cell Membrane / Q000032:analysis; D010450:Endopeptidases / Q000502:physiology*; D005165:Factor V / Q000502:physiology; D015943:Factor Va; D015951:Factor Xa; D006801:Humans; D010447:Peptide Hydrolases / Q000502:physiology*; D010741:Phospholipases A / Q000494:pharmacology; D054467:Phospholipases A2; D010743:Phospholipids / Q000502:physiology; D012697:Serine Endopeptidases / Q000502:physiology; D012910:Snake Venoms / Q000502:physiology; D014757:Viper Venoms / Q000502:physiology
D001779:Blood Coagulation Factors; D010743:Phospholipids; D012910:Snake Venoms; D014757:Viper Venoms; C049426:platelet procoagulant factor; D015943:Factor Va; D005165:Factor V; D010741:Phospholipases A; D054467:Phospholipases A2; D010450:Endopeptidases; D010447:Peptide Hydrolases; D012697:Serine Endopeptidases; D015951:Factor Xa; C020097:ecarin