Identification of a heparin-binding protein using monoclonal antibodies that block heparin binding to porcine aortic endothelial cells.
The binding of heparin or heparan sulphate to a variety of cell types results in specific changes in cell function. Endothelial cells treated with heparin alter their synthesis of heparan sulphate proteoglycans and extracellular matrix proteins. In order to identify a putative endothelial cell heparin receptor that could be involved in heparin signalling, anti-(endothelial cell) monoclonal antibodies that significantly inhibit heparin binding to endothelial cells were prepared. Four of these antibodies were employed in affinity-chromatographic isolation of a heparin-binding protein from detergent-solubilized endothelial cells. The heparin-binding protein isolated from porcine aortic endothelial cells using four different monoclonal antibodies has an M(r) of 45,000 assessed by SDS/PAGE. The 45,000-M(r) heparin-binding polypeptide is isolated as a multimer. The antibody-isolated protein binds to heparin-affinity columns as does the pure 45,000-M(r) polypeptide, consistent with its identification as a putative endothelial heparin receptor.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D001013:Aorta, Thoracic; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D005260:Female; D006493:Heparin / Q000378:metabolism*; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011485:Protein Binding / Q000187:drug effects; D011956:Receptors, Cell Surface / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D013552:Swine
D000911:Antibodies, Monoclonal; D011956:Receptors, Cell Surface; C037680:heparin receptor; D006493:Heparin