Abstract
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.
Copyright 2001 Wiley-Liss, Inc.
Publication types
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Animals
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Binding Sites
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Calcium / metabolism
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Cattle
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Cell Adhesion
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Cells, Cultured
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Chondroitin Sulfate Proteoglycans / chemistry*
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Dose-Response Relationship, Drug
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E-Selectin / chemistry*
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E-Selectin / isolation & purification
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E-Selectin / metabolism
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Endothelium, Vascular / metabolism
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Enzyme-Linked Immunosorbent Assay / methods*
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Epithelial Cells / metabolism
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Female
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Glycosaminoglycans / metabolism
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HL-60 Cells
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Heparin / analogs & derivatives
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Heparin / chemistry*
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Heparin / metabolism
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Heparin / pharmacology
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Humans
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Ligands
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Mammary Glands, Animal / metabolism
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Membrane Glycoproteins / chemistry
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Membrane Glycoproteins / isolation & purification
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Models, Biological
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Oligosaccharides / metabolism
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P-Selectin / metabolism
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Polylysine / chemistry
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Protein Binding
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Protein Structure, Tertiary
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Proteoglycans / chemistry*
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Proteoglycans / isolation & purification
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Sialyl Lewis X Antigen
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Syndecan-1
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Syndecans
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Trypsin / pharmacology
Substances
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Chondroitin Sulfate Proteoglycans
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E-Selectin
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Glycosaminoglycans
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Ligands
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Membrane Glycoproteins
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Oligosaccharides
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P-Selectin
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Proteoglycans
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SDC1 protein, human
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Sialyl Lewis X Antigen
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Syndecan-1
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Syndecans
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heparin proteoglycan
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Polylysine
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Heparin
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Trypsin
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Calcium