Peptides derived from a sequence within beta 3 integrin bind to platelet alpha IIb beta 3 (GPIIb-IIIa) and inhibit ligand binding.

Peptides derived from a sequence within the loop structure of human platelet glycoprotein (GP) IIIa (integrin beta 3) were previously shown to inhibit fibrinogen binding to purified GPIIb-IIIa. In this study a series of peptides based on the GPIIIa sequence 211-221 (SVSRNRDAPEG) was synthesized. The most active peptide was determined to be RNRDA, and its inhibitory potency was 4-fold greater (IC50 = 4.8 microM) than that of SVSRNRDAPEG. These GPIIIa peptides also inhibited the binding of two monoclonal antibodies, pl-55 and PAC-1, which are directed against the activated conformer of GPIIb-IIIa. To determine whether these peptides bound directly to GPIIb-IIIa, an affinity matrix was prepared by coupling RNRDAPEGC to Sepharose. Fibrinogen or purified GPIIb-IIIa was applied to the affinity column. Only GPIIb-IIIa was retained on the column, and it could be specifically eluted by GPIIIa peptide or RGDV but not by an irrelevant peptide. Additionally, we observed that the binding of GPIIIa peptides to purified GPIIb-IIIa induced exposure of a neoepitope on GPIIb that was recognized by the monoclonal antibody pl-80. These data suggest that sequences within the loop structure of GPIIIa can interact with the ligand binding domain of GPIIb-IIIa. Thus, this GPIIIa domain may be involved in regulating the accessibility of ligands to GPIIb-IIIa following platelet activation.