Heparan sulfate-mediated binding of epithelial cell surface proteoglycan to thrombospondin.
Purified NMuMG mouse mammary epithelial cell surface proteoglycan (PG), a membrane-intercalated core protein bearing both heparan sulfate and chondroitin sulfate glycosaminoglycan (GAG) chains, binds to a thrombospondin (TSP) affinity column and is eluted by a salt gradient. Double immunofluorescence microscopy demonstrates extensive co-localization of bound exogenous TSP and cells bearing exposed cell surface PG at their apical surface. The binding, as assayed by both methods, is heparitinase-sensitive, but not chondroitinase-sensitive. Alkali-released heparan sulfate chains bind to a TSP affinity column, similarly to native PG, whereas the chrondroitin sulfate chains do not. Core protein does not bind to TSP. These results indicate that NMuMG cells bind TSP via their surface PG and that the binding is mediated by the heparan sulfate chains.
Sun
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Department of Medicine, University of Wisconsin, Madison 53706.
Mosher
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Rapraeger
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D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D002846:Chromatography, Affinity; D004848:Epithelium / Q000032:analysis; D006023:Glycoproteins / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000302:isolation & purification*; D006497:Heparan Sulfate / Q000302:isolation & purification* / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000032:analysis; D051379:Mice; D011485:Protein Binding; D011509:Proteoglycans / Q000302:isolation & purification* / Q000378:metabolism; D019699:Thrombospondins
D006023:Glycoproteins; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans; D019699:Thrombospondins