Selective induction of a glycoprotein IIIa ligand-induced binding site by fibrinogen and von Willebrand factor.
Ligand-induced binding sites (LIBS) are neoantigenic regions of glycoprotein (GP)IIb-IIIa that are exposed upon interaction of the receptor with the ligand fibrinogen or the ligand recognition sequence (RGDS). LIBS have been suggested to contribute to postreceptor occupancy events such as full-scale platelet aggregation, adhesion to collagen, and clot retraction. This study examined the induction requirements of a GPIIIa LIBS with regard to ligand specificity. Through the use of the anti-LIBS D3, we report that this complex-activating antibody induces fibrinogen- and von Willebrand factor-binding to GPIIb-IIIa on intact platelets. Bound ligand was detected by flow cytometric analysis and platelet aggregation assays. These bound ligands increased the number of D3-binding sites and altered the affinity of D3 for GPIIb-IIIa on platelets. In contrast, activation of platelet GPIIb-IIIa by D3 did not increase the binding of another RGD-containing ligand, vitronectin. Furthermore, bound vitronectin on thrombin-stimulated platelets did not cause the expression of the D3 LIBS epitope. We conclude direct activation of GPIIb-IIIa in the absence of platelet activation results in selective ligand interaction and that D3 LIBS induction requires the binding of the multivalent ligands, fibrinogen or von Willebrand factor. Thus, the region of GPIIIa recognized by D3 may be an important regulatory domain in ligand-receptor interactions that directly mediate platelet aggregation.
Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda MD, USA.
T H
TH
Mondoro
C D
CD
Wall
M M
MM
White
L K
LK
Jennings
D000818:Animals; D001665:Binding Sites; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D008024:Ligands; D051379:Mice; D009842:Oligopeptides / Q000494:pharmacology; D010974:Platelet Aggregation; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000737:chemistry / Q000187:drug effects* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects*; D013917:Thrombin / Q000494:pharmacology; D019096:Vitronectin / Q000378:metabolism; D014841:von Willebrand Factor / Q000378:metabolism / Q000494:pharmacology*
D008024:Ligands; D009842:Oligopeptides; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D019096:Vitronectin; D014841:von Willebrand Factor; D005340:Fibrinogen; C046617:arginyl-glycyl-aspartyl-serine; D013917:Thrombin