The fibrinogen RIBS-I epitope (gamma 373-385) appears proximate to the gamma 408-411 adhesive domain but is not involved in interaction between receptor-bound or surface-adsorbed fibrinogen and platelet GPIIbIIIa

The carboxyl terminus of the fibrinogen (Fg) gamma chain (gamma 400-411) is necessary and sufficient to support platelet aggregation and adhesion. How ever, a monoclonal antibody (mAb) to the Fg RIBS-I epitope (gamma 373-385), the anti-Fg-RIBS-I, which binds only to platelet-bound or surface-adsorbed Fg but not soluble Fg, inhibits platelet aggregation. In this study, we sh owed that this same antibody also inhibits the adhesion of platelets to Fg- coated polystyrene beads. We then investigated the mechanisms by which the anti-Fg-RIBS-I antibody inhibits platelet aggregation and adhesion. The Fg RIBS-I epitope does not interact with platelet GPIIbIIIa, since recombinant Fg missing the last four amino acids, the Ala-Gly-Asp-Val, on the carboxyl terminus of its gamma chains supports neither platelet aggregation nor adh esion to surfaces, nor GPIIbIIIa binding, while it binds anti-Fg-RIBS-I nor mally. Purified, soluble GPIIbIIIa (265 kDa) inhibits the binding of both t he anti-Fg-RIBS-I and 4A5 (a mAb specific to gamma 408-411 of Fg); however, peptide G13 (1.5 kDa), corresponding to the Fg gamma chain binding domain on GPIIba (GPIIb300-312), only inhibits the binding of 4A5, and does not af fect the binding of the anti-Fg-RIBS-I to Fg. The anti-Fg-RIBS-I reduces th e on-rate of the 4A5 binding to Fg with no measurable changes in the dissoc iation of the Fg-bound 4A5. These data indicate that the inhibition of plat elet aggregation and adhesion by the anti-Fg-RIBS-I antibody is due to the steric hindrance of the Fg gamma 400-411 to platelet GPIIbIIIa. Thus the Fg RIBS-I epitope (gamma 373-385) does not appear to be involved in direct in teraction with platelet GPIIbIIIa, leaving the gamma 408-411 of Fg as the s ole domain mediating platelet aggregation and adhesion. (C) 1998 Elsevier S cience B.V. All rights reserved.