Activation-independent exposure of the GPIIb-IIIa fibrinogen receptor.
Following platelet activation, surface receptors for fibrinogen are exposed. On the activated platelet, glycoprotein IIb-IIIa (GPIIb-IIIa) serves as the receptor for fibrinogen. However, the molecular mechanisms which regulate GPIIb-IIIa fibrinogen receptor exposure are unknown. D3GP3 is an IgG1, kappa monoclonal antibody which is specific for glycoprotein IIIa (GPIIIa). The binding of D3GP3 to GPIIIa, in intact GPIIb-IIIa complexes, induces fibrinogen binding and platelet aggregation. To determine if D3GP3 binding to GPIIIa directly caused the exposure of fibrinogen receptors or, secondarily, due to stimulus response coupling, platelet activation parameters were monitored following the addition of D3GP3 to platelets suspensions. D3GP3 binding did not induce detectable Ca++ mobilization, protein phosphorylation or activation of the pertussis toxin sensitive G-protein subunit alpha-41. Further, D3GP3-induced aggregation was not blocked by PGE1, aspirin, apyrase or the combination of all three reagents. Scanning electron microscopy of D3GP3-induced aggregates demonstrated that the aggregates were composed of discoid platelets. These data suggest that the binding of D3GP3 to GPIIIa induced a conformational change in GPIIb-IIIa such that the fibrinogen receptor was exposed in an activation-independent fashion. This provides evidence that conformational changes in the GPIIb-IIIa complex can result in the transformation of the complex to the high affinity binding competent state.
Kouns
W C
WC
Department of Medicine, University of Tennessee, Memphis 38163.
Jennings
L K
LK
D000911:Antibodies, Monoclonal / Q000378:metabolism; D000937:Antigen-Antibody Reactions; D001792:Blood Platelets / Q000378:metabolism* / Q000648:ultrastructure; D002118:Calcium / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D008855:Microscopy, Electron, Scanning; D037342:Pertussis Toxin; D010766:Phosphorylation; D015539:Platelet Activation; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011487:Protein Conformation; D011499:Protein Processing, Post-Translational; D015290:Second Messenger Systems / Q000187:drug effects; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D010980:Platelet Membrane Glycoproteins; D010566:Virulence Factors, Bordetella; D005340:Fibrinogen; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D002118:Calcium