Selective binding of the factor VIII/von Willebrand factor protein to human platelets.
The factor VIII/von Willebrand factor protein was radiolabeled after modification by galactose oxidase and reduction with tritiated potassium borohydride. This mild efficient method for labeling resulted in retention of over 90% of the biologic activities of the factor VIII/von Willebrand factor protein. Binding of this protein to platelets was found to be specific, and binding sites could be saturated in the presence of ristocetin. However, binding was highly dependent on ristocetin concentration, as the number of human factor VIII/von Willebrand factor molecules bound per platelet was a function of the ristocetin concentration. At a ristocetin concentration of 0.55 mg/ml, each platelet binds approximately 11,000 factor VIII/von Willebrand factor molecules per platelet. Scatchard analysis of the concentration-dependent binding sites yielded a hyperbolic plot that appeared to be related to the existence of two classes of binding sites. The higher affinity class had a Kd of 3.7 x 10(-10) M 3500 sites/platelet, while the lower affinity class had a Kd of 2.35 x 10(-9) M and a capacity of 7500 sites/platelet. As with ristocetin-induced platelet agglutination, the carbohydrate content plays a significant role in the binding of the factor VIII/von Willebrand factor protein to the platelet.
D000941:Antigens; D001665:Binding Sites; D001779:Blood Coagulation Factors; D001792:Blood Platelets / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D005169:Factor VIII; D005340:Fibrinogen / Q000494:pharmacology; D005692:Galactose Oxidase / Q000494:pharmacology; D005696:Galactosidases / Q000494:pharmacology; D006801:Humans; D009439:Neuraminidase / Q000494:pharmacology; D010974:Platelet Aggregation; D012310:Ristocetin / Q000494:pharmacology; D012794:Sialic Acids; D014841:von Willebrand Factor
D000941:Antigens; D001779:Blood Coagulation Factors; D002352:Carrier Proteins; D012794:Sialic Acids; D014841:von Willebrand Factor; D012310:Ristocetin; D005169:Factor VIII; D005340:Fibrinogen; D005692:Galactose Oxidase; D005696:Galactosidases; D009439:Neuraminidase