Binding of a large chondroitin sulfate/dermatan sulfate proteoglycan, versican, to L-selectin, P-selectin, and CD44.
Here we show that a large chondroitin sulfate proteoglycan, versican, derived from a renal adenocarcinoma cell line ACHN, binds L-selectin, P-selectin, and CD44. The binding was mediated by the interaction of the chondroitin sulfate (CS) chain of versican with the carbohydrate-binding domain of L- and P-selectin and CD44. The binding of versican to L- and P-selectin was inhibited by CS B, CS E, and heparan sulfate (HS) but not by any other glycosaminoglycans tested. On the other hand, the binding to CD44 was inhibited by hyaluronic acid, chondroitin (CH), CS A, CS B, CS C, CS D, and CS E but not by HS or keratan sulfate. A cross-blocking study indicated that L- and P-selectin recognize close or overlapping sites on versican, whereas CD44 recognizes separate sites. We also show that soluble L- and P-selectin directly bind to immobilized CS B, CS E, and HS and that soluble CD44 directly binds to immobilized hyaluronic acid, CH, and all the CS chains examined. Consistent with these results, structural analysis showed that versican is modified with at least CS B and CS C. Thus, proteoglycans sufficiently modified with the appropriate glycosaminoglycans should be able to bind L-selectin, P-selectin, and/or CD44.
Kawashima
H
H
Department of Bioregulation, Biomedical Research Center, Osaka University Graduate School of Medicine 2-2, Yamada-oka, Suita 565-0871, Japan.
Hirose
M
M
Hirose
J
J
Nagakubo
D
D
Plaas
A H
AH
Miyasaka
M
M
D000818:Animals; D019921:Biotinylation; D015153:Blotting, Western; D002807:Chondroitin / Q000494:pharmacology; D019765:Chondroitin ABC Lyase / Q000494:pharmacology; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism*; D002809:Chondroitin Sulfates / Q000378:metabolism*; D002810:Chondroitinases and Chondroitin Lyases / Q000494:pharmacology; D003432:Cross-Linking Reagents / Q000494:pharmacology; D018076:DNA, Complementary / Q000378:metabolism; D003871:Dermatan Sulfate / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D018960:Hyaluronan Receptors / Q000378:metabolism*; D006820:Hyaluronic Acid / Q000494:pharmacology; D007632:Keratan Sulfate / Q000494:pharmacology; D007700:Kinetics; D019041:L-Selectin / Q000378:metabolism*; D037181:Lectins, C-Type; D050356:Lipid Metabolism; D051379:Mice; D019007:P-Selectin / Q000378:metabolism*; D011485:Protein Binding; D011509:Proteoglycans / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured; D053675:Versicans
D011508:Chondroitin Sulfate Proteoglycans; D003432:Cross-Linking Reagents; D018076:DNA, Complementary; D006025:Glycosaminoglycans; D018960:Hyaluronan Receptors; D037181:Lectins, C-Type; D019007:P-Selectin; D011509:Proteoglycans; C508322:VCAN protein, human; C508323:Vcan protein, mouse; D019041:L-Selectin; D053675:Versicans; D003871:Dermatan Sulfate; D006820:Hyaluronic Acid; D002807:Chondroitin; D002809:Chondroitin Sulfates; D007632:Keratan Sulfate; D002810:Chondroitinases and Chondroitin Lyases; D019765:Chondroitin ABC Lyase