Fibrinogen is a ligand for integrin alpha5beta1 on endothelial cells.
Previous studies have shown that fibrinogen can associate with endothelial cells via an Arg-Gly-Asp (RGD) recognition specificity. In the present study, we have characterized the specificity of fibrinogen binding to endothelial cells under different cation conditions. Fibrinogen binding to suspended endothelial cells was selectively supported by Mn2+ and was suppressed by Ca2+. The Mn2+-supported interaction was completely inhibited by RGD peptides but not by alphavbeta3 blocking monoclonal antibodies. In contrast, the interaction was completely blocked by two alpha5beta1 monoclonal antibodies. This interaction was not mediated by fibronectin bound to the integrin; could be demonstrated with purified alpha5beta1; and also was observed with a second alpha5beta1-bearing cell type, platelets. The binding of fibrinogen to alpha5beta1 on endothelial cells in the presence of Mn2+ was time-dependent, specific, saturable, and of high affinity (Kd = 65 nM). By employing anti-peptide monoclonal antibodies, the carboxyl-terminal RGD sequence at Aalpha 572-574 was implicated in fibrinogen recognition by alpha5beta1. Two circumstances were identified in which alpha5beta1 interacted with fibrinogen in the presence of Ca2+: when the receptor was activated with monoclonal antibody (8A2) or when the fibrinogen was presented as an immobilized substratum. These results identify fibrinogen as a ligand for alpha5beta1 on endothelial and other cells, an interaction which may have broad biological implications.
Suehiro
K
K
Joseph J. Jacobs Center for Thrombosis and Vascular Biology, Department of Molecular Cardiology, The Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
Gailit
J
J
Plow
E F
EF
D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D017469:Receptors, Fibronectin / Q000378:metabolism*
D008024:Ligands; D017469:Receptors, Fibronectin; D005340:Fibrinogen