Platelet aggregation by fibrinogen polymers crosslinked across the E domain.
There is evidence that platelet interactions with artificial surfaces are mediated by plasma proteins, especially fibrinogen, adsorbed on the surfaces. Multiple site interactions between fibrinogen molecules adsorbed in high concentration and receptors in the unactivated platelet may be sufficient for platelet adhesion and subsequent activation. To examine this hypothesis, we prepared soluble polymers of fibrinogen. Polymers produced by interaction of fibrinogen with Fab'2 fragments of antibodies against fibrinogen's E (central) domain (Fg-Fab'2(E] induced, in gel-filtered platelets, aggregation and serotonin release, which were blocked by monoclonal antibodies against the GPIIb/IIIa complex, by Fab fragments against the D domain, and by metabolic inhibitors; aggregation was attenuated but not abolished by enzymatic removal of ADP (with CP/CPK) or by blockage of ADP binding sites (with FSBA), and when secretion was inhibited by aspirin. Fg-Fab'2(E) also induced a dose-dependent elevation in cytoplasmic Ca2+ (measured by Aequorin luminescence) which was attenuated by CP/CPK and by FSBA, and was eliminated by metabolic inhibitors and by anti-IIb/IIIa antibody. Fibrinogen complexes crosslinked with dimethylsuberimidate or Factor XIII neither aggregated gel-filtered platelets nor inhibited platelet aggregation by ADP and fibrinogen, probably because of inaccessibility of lysine residues in the D (terminal) domain of fibrinogen, which are thought to be required for platelet binding. Thus, soluble complexes of fibrinogen having multiple available platelet receptor recognition sites activate gel-filtered platelets and may provide a useful model for platelet-surface interactions mediated by adsorbed fibrinogen.
D000244:Adenosine Diphosphate / Q000494:pharmacology; D002118:Calcium / Q000032:analysis; D003432:Cross-Linking Reagents; D003593:Cytoplasm / Q000032:analysis; D005338:Fibrin Fibrinogen Degradation Products / Q000502:physiology*; D005340:Fibrinogen / Q000494:pharmacology*; D006801:Humans; D010725:Phosphocreatine / Q000494:pharmacology; D010974:Platelet Aggregation; D011108:Polymers; D012701:Serotonin / Q000378:metabolism
D003432:Cross-Linking Reagents; D005338:Fibrin Fibrinogen Degradation Products; D011108:Polymers; C018543:fibrinogen fragment E; D010725:Phosphocreatine; D012701:Serotonin; D000244:Adenosine Diphosphate; D005340:Fibrinogen; D002118:Calcium