title: Chemical cross-linking of arginyl-glycyl-aspartic acid peptides to an adhesion receptor on platelets.
abstract: A chemical cross-linking approach has been used to characterize the interaction of platelets with small peptides of 7 and 14 residues containing the arginyl-glycyl-aspartic acid (RGD) sequence recognized by a variety of cellular adhesion receptors. The radioiodinated peptides were bound to platelets, and chemical cross-linking was attained by subsequent addition of bifunctional reagents. Three different cross-linking reagents coupled the RGD-containing peptides to platelet membrane glycoprotein IIb-IIIa (GPIIb-IIIa), and both subunits of this platelet membrane glycoprotein became radiolabeled with the RGD peptides. Platelet stimulation with agonists including thrombin, phorbol myristrate acetate, and ADP increased the extent of cross-linking by predominantly enhancing the coupling of the RGD peptides to the GPIIIa subunit. Cross-linking of the labeled RGD peptides to GPIIb and GPIIIa on stimulated and nonstimulated platelets exhibited structural specificity and was inhibited by excess nonlabeled RGD peptides. The interactions were inhibited by nonlabeled RGD peptides and a peptide with an amino acid sequence corresponding to the carboxyl terminus of the gamma chain of fibrinogen but less effectively by an arginyl-glycyl-glutamic acid peptide. Cross-linking of the RGD peptides to GPIIb-IIIa was divalent ion-dependent and, on stimulated platelets, was inhibited by the adhesive proteins fibrinogen and fibronectin, but not by albumin. These results indicate that the RGD-binding sites on platelets reside in close proximity to both subunits of GPIIb-IIIa and that platelet stimulation alters the topography of these sites such that the peptides become more efficiently cross-linked to GPIIIa.
authors.affiliation: Department of Immunology, Research Institute of Scripps Clinic, La Jolla, California 92037.
authors.forename: S E
authors.initials: SE
authors.lastname: D'Souza
authors.forename: M H
authors.initials: MH
authors.lastname: Ginsberg
authors.forename: S C
authors.initials: SC
authors.lastname: Lam
authors.forename: E F
authors.initials: EF
authors.lastname: Plow
mesh_terms: D000595:Amino Acid Sequence; D000954:Antigens, Surface / Q000378:metabolism*; D001120:Arginine; D001224:Aspartic Acid; D001792:Blood Platelets / Q000378:metabolism*; D015815:Cell Adhesion Molecules; D003432:Cross-Linking Reagents; D005353:Fibronectins / Q000097:blood; D005998:Glycine; D006801:Humans; D009842:Oligopeptides / Q000097:blood* / Q000138:chemical synthesis; D010973:Platelet Adhesiveness; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D013917:Thrombin / Q000502:physiology
chemical_list: D000954:Antigens, Surface; D015815:Cell Adhesion Molecules; D003432:Cross-Linking Reagents; D005353:Fibronectins; D009842:Oligopeptides; D010980:Platelet Membrane Glycoproteins; D001224:Aspartic Acid; D001120:Arginine; D013917:Thrombin; D005998:Glycine