Heparan sulfate and chondroitin sulfate proteoglycans inhibit E-selectin binding to endothelial cells.
E-selectin is a cell adhesion molecule involved in the initial rolling and adhesion of leukocytes to the endothelium during inflammation. In addition, in vitro studies have suggested that an interaction between E-selectin and binding sites such as sialyl Lewis X-containing oligosaccharides on endothelial cells may be important for angiogenesis. In order to investigate the binding of E-selectin to endothelial cells, we developed an ELISA assay using chimeric E-selectin-Ig molecules and endothelial cells fixed on poly-L-lysine coated plates. Our results indicate that E-selectin-Ig binds to both bovine capillary endothelial cells and human dermal microvascular endothelial cells in a calcium-dependent and saturable manner. The binding is inhibited markedly by heparin and by syndecan-1 ectodomain, and moderately by chondroitin sulfate, but not by sialyl Lewis X-containing oligosaccharides. These results suggest that heparan sulfate and chondroitin sulfate proteoglycans on endothelial cells are potential ligands for E-selectin.
Luo
J
J
Surgical Research Laboratory, Children's Hospital, 300 Longwood Ave., Boston, MA 02115, USA.
Kato
M
M
Wang
H
H
Bernfield
M
M
Bischoff
J
J
D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002417:Cattle; D002448:Cell Adhesion; D002478:Cells, Cultured; D011508:Chondroitin Sulfate Proteoglycans / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D019040:E-Selectin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D004730:Endothelium, Vascular / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004847:Epithelial Cells / Q000378:metabolism; D005260:Female; D006025:Glycosaminoglycans / Q000378:metabolism; D018922:HL-60 Cells; D006493:Heparin / Q000031:analogs & derivatives / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D008024:Ligands; D008321:Mammary Glands, Animal / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000302:isolation & purification; D008954:Models, Biological; D009844:Oligosaccharides / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism; D011107:Polylysine / Q000737:chemistry; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000737:chemistry* / Q000302:isolation & purification; D000080506:Sialyl Lewis X Antigen; D053668:Syndecan-1; D053667:Syndecans; D014357:Trypsin / Q000494:pharmacology
D011508:Chondroitin Sulfate Proteoglycans; D019040:E-Selectin; D006025:Glycosaminoglycans; D008024:Ligands; D008562:Membrane Glycoproteins; D009844:Oligosaccharides; D019007:P-Selectin; D011509:Proteoglycans; C508241:SDC1 protein, human; D000080506:Sialyl Lewis X Antigen; D053668:Syndecan-1; D053667:Syndecans; C036090:heparin proteoglycan; D011107:Polylysine; D006493:Heparin; D014357:Trypsin; D002118:Calcium