Involvement of fibronectin, Von Willebrand factor, and fibrinogen in platelet interaction with solid substrata.
The proteins fibronectin (FN), Von Willebrand factor (VWF), and fibrinogen are believed to play a role in platelet function. They are distributed between the plasma and the platelet pool in the resting state and undergo redistribution upon platelet activation. We have studied their expression on the surface of the platelet and their mobilization following platelet binding to substrata. For the purpose of studying protein expression on the surface of intact platelets either adherent to a substratum or in suspension, the enzyme-linked immunosorbent assay (ELISA) was elaborated and modified. Using this technique as well as immunofluorescence, we found that antiserum raised against carefully washed human platelets recognized FN, VWF, and fibrinogen as well as platelet surfaces. However, specific antisera against these three proteins failed to bind to the surface of unactivated gel-filtered platelets. When gel-filtered platelets were exposed to plastic or fibrillar collagen, they adhered and spread. Such platelets did bind antibodies against FN, VWF, and fibrinogen, Moreover, when the adherent platelets were incubated with FN or with VWF in the absence of ristocetin, they bound these proteins in a concentration-dependent fashion. The patterns of the bound proteins were not similar, suggesting a different spatial distribution of binding sites. These findings indicate that platelet activation by adhesion to substrata mobilize both endogenous and exogenous pools of these proteins, thereby making them surface associated and probable participants in further binding properties of the activated platelet.
D000328:Adult; D000818:Animals; D000954:Antigens, Surface; D001779:Blood Coagulation Factors / Q000378:metabolism*; D001792:Blood Platelets / Q000502:physiology*; D003094:Collagen / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005110:Extracellular Space; D005340:Fibrinogen / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D010974:Platelet Aggregation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D011817:Rabbits; D014841:von Willebrand Factor / Q000378:metabolism*
D000954:Antigens, Surface; D001779:Blood Coagulation Factors; D005353:Fibronectins; D014841:von Willebrand Factor; D005340:Fibrinogen; D003094:Collagen