Studies on the mechanism of expression of secreted fibrinogen on the surface of activated human platelets.
Affinity purified anti-fibrinogen (anti-Fg) Fab fragments were used to study the mechanism of expression of alpha-granule fibrinogen on activated platelets. Low amounts of the radiolabeled anti-Fg Fab bound to unstimulated or adenosine diphosphate (ADP)-stimulated cells. They readily bound to platelets stimulated with collagen, alpha-thrombin or gamma-thrombin in the presence of divalent cations. At 1 n mol/L alpha-thrombin or 25 nmol/L gamma-thrombin, platelet fibrinogen was expressed on the surface of the cells notwithstanding the presence of AP-2, a monoclonal antibody to the glycoprotein (GP) IIb-IIIa complex, or the synthetic peptides Arg-Gly-Asp-Ser and gamma 400-411, all substances that prevented the binding of plasma fibrinogen to platelets. These results suggest that platelet fibrinogen may interact with its receptors during its translocation from the alpha-granules to the plasma membrane and, thus, not occupy the same sites as those available for plasma fibrinogen on the surface of the cell. Furthermore, we found that platelet fibrinogen was expressed on the thrombin-stimulated platelets of a Glanzmann's thrombasthenia variant that failed to bind plasma fibrinogen. Normal platelets stimulated with 5 nmol/L alpha-thrombin bound increased amounts of the anti-fg Fab, the additional expression being inhibited by the anti-GP IIb-IIIa monoclonal antibody or by Gly-Pro-Arg-Pro, an inhibitor of fibrin polymer formation. This suggests that rebinding to externally located GP IIb-IIIa complexes becomes important once fibrin is formed.
Legrand
C
C
Laboratoire d'Hémostase et de Thrombose Expérimentale, Hôpital Lariboisière, Paris, France.
Dubernard
V
V
Nurden
A T
AT
D001666:Binding Sites, Antibody; D001792:Blood Platelets / Q000378:metabolism / Q000502:physiology*; D002462:Cell Membrane / Q000378:metabolism / Q000502:physiology; D005337:Fibrin / Q000378:metabolism; D005340:Fibrinogen / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D010974:Platelet Aggregation; D011956:Receptors, Cell Surface / Q000032:analysis; D013915:Thrombasthenia / Q000097:blood
D007140:Immunoglobulin Fab Fragments; D011956:Receptors, Cell Surface; D005337:Fibrin; D005340:Fibrinogen