Solid-phase von Willebrand factor contains a conformationally active RGD motif that mediates endothelial cell adhesion through the alpha v beta 3 receptor.
The interaction of von Willebrand factor (vWF) with the alpha v beta 3 integrin of human umbilical vein endothelial cells is dependent on the RGD sequence present at residues 1744-1746 of the mature vWF subunit. We compared vWF and its two dimeric fragments, SpIII (residues 1-1365) and SpII (residues 1366-2050), as adhesion substrates. Solid-phase vWF and SpII supported endothelial cell adhesion, whereas SpIII, which contains the glycoprotein (GP) Ib binding domain, did not. Soluble SpII inhibited adhesion to immobilized ligands, whereas soluble vWF did not, suggesting that exposure of the cell attachment domain involves a conformational modification of vWF. Dendroaspin and albolabrin, two RGD-containing peptides of the disintegrin family, were potent inhibitors of cell adhesion to vWF (IC50 approximately 15 nmol/L). Complete inhibition of endothelial cell adhesion to vWF was obtained in the presence of F(ab')2 of monoclonal antibody 9 to vWF, which blocks vWF binding to platelet GPIIb/IIIa. In contrast, monoclonal antibody 713 to vWF, which blocks its binding to platelet GPIb, did not inhibit cell adhesion to vWF. These results indicate that endothelial cell adhesion to vWF is mediated by an RGD-dependent interaction with alpha v beta 3, but does not seem to involve a GPIb-like receptor, and show the importance of the conformation of the RGD sequence.
Denis
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INSERM U. 143, Hôpital de Bicêtre, Paris, France.
Williams
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JA
Lu
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Meyer
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Baruch
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D000595:Amino Acid Sequence; D000906:Antibodies; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002448:Cell Adhesion / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D003571:Cytochalasin B / Q000494:pharmacology; D004546:Elapid Venoms / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000187:drug effects / Q000502:physiology*; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology; D016023:Integrins / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000032:analysis* / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology; D010455:Peptides / Q000138:chemical synthesis / Q000494:pharmacology; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D016024:Receptors, Cytoadhesin / Q000378:metabolism*; D019030:Receptors, Vitronectin; D012910:Snake Venoms; D014471:Umbilical Veins; D014841:von Willebrand Factor / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D004546:Elapid Venoms; D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D016023:Integrins; D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; D010975:Platelet Aggregation Inhibitors; D016024:Receptors, Cytoadhesin; D019030:Receptors, Vitronectin; D012910:Snake Venoms; D014841:von Willebrand Factor; C064595:albolabrin; C074772:mambin; D003571:Cytochalasin B; C047981:arginyl-glycyl-aspartic acid