title: Functional analysis of a recombinant glycoprotein Ia/IIa (Integrin alpha(2)beta(1)) I domain that inhibits platelet adhesion to collagen and endothelial matrix under flow conditions.
abstract: The interaction of platelets with collagen plays an important role in primary hemostasis. Glycoprotein Ia/IIa (GPIa/IIa, integrin alpha(2)beta(1)) is a major platelet receptor for collagen. The binding site for collagen has been mapped to the I domain within the alpha(2) subunit (GPIa). In order to assess the role of the alpha(2)-I domain structure in GPIa/IIa binding to collagen, a recombinant I domain (amino acids 126-337) was expressed in Escherichia coli. The alpha(2)-I protein bound human types I and III collagen in a saturable and divalent cation-dependent manner and was blocked by the alpha(2)beta(1) function blocking antibody 6F1. The alpha(2)-I protein inhibited collagen-induced platelet aggregation (IC(50) = 600 nM). Unexpectedly, 6F1, an antibody that fails to inhibit platelet aggregation in platelet-rich plasma, blocked the inhibitory effect of the alpha(2)-I protein. The alpha(2)-I protein was able to prevent platelet adhesion to a collagen surface exposed to flowing blood under low shear stress. Interestingly, it inhibited platelet adhesion to extracellular matrix at high shear stress. These results, taken together, provide firm evidence that GPIa/IIa directly mediates the first contact of platelets with collagen under both stirring and flow conditions.
authors.affiliation: Hematology Division, Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.
authors.forename: D
authors.initials: D
authors.lastname: Estavillo
authors.forename: A
authors.initials: A
authors.lastname: Ritchie
authors.forename: T G
authors.initials: TG
authors.lastname: Diacovo
authors.forename: M A
authors.initials: MA
authors.lastname: Cruz
mesh_terms: D001667:Binding, Competitive; D002413:Cations, Divalent / Q000494:pharmacology; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003094:Collagen / Q000378:metabolism*; D004492:Edetic Acid / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000502:physiology*; D004926:Escherichia coli; D005109:Extracellular Matrix / Q000502:physiology*; D006801:Humans; D016023:Integrins / Q000737:chemistry* / Q000502:physiology*; D007700:Kinetics; D010446:Peptide Fragments / Q000737:chemistry / Q000494:pharmacology*; D010973:Platelet Adhesiveness / Q000187:drug effects / Q000502:physiology*; D010974:Platelet Aggregation / Q000187:drug effects*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D038981:Receptors, Collagen; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D014471:Umbilical Veins
chemical_list: D002413:Cations, Divalent; D016023:Integrins; D010446:Peptide Fragments; D010975:Platelet Aggregation Inhibitors; D038981:Receptors, Collagen; D011994:Recombinant Proteins; D003094:Collagen; D004492:Edetic Acid