Decorin endocytosis: structural features of heparin and heparan sulphate oligosaccharides interfering with receptor binding and endocytosis.
Receptor-mediated endocytosis of decorin depends on its core-protein-mediated interaction with a 51 kDa membrane protein, which, in addition to its core-protein-binding site, carries a binding site for glycosaminoglycan chains. Membrane-associated heparan sulphate as well as heparin are known to have an inhibitory effect on decorin endocytosis by cultured skin fibroblasts. In this study, structural features of both glycosaminoglycans required for binding to the 51 kDa protein and for inhibiting decorin endocytosis, were investigated. Upon digestion of [(3)H]glucosamine-labelled heparan sulphate with heparinase III, dodeca- and higher saccharides were able to interact with the receptor protein. In comparison with unbound fragments of the same size, bound fragments were enriched in N-sulphated disaccharides carrying one or two sulphate ester groups. Using heparinase III-generated fragments from [(35)S]sulphate-labelled heparan sulphate chains, binding of fragments as small as octasaccharides could be detected. Competition experiments between dermatan sulphate and chemically modified heparin revealed that N- and 6-O-sulphation of glucosamine residues are important structural elements for binding to the receptor, whereas iduronate-2-O-sulphate groups contribute to binding only to a limited extent. However, with respect to the inhibition of decorin endocytosis, 2-O-desulphation had a quantitatively similar effect to 6-O-desulphation. Furthermore, for maximal inhibition of decorin endocytosis, longer fragments were required than for binding to the receptor. Thus, it appears that heparin/heparan sulphate has to interact with additional component(s) for effective inhibition of decorin uptake.
Hausser
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Institute of Physiological Chemistry, University of Münster, D-48129 Münster, Germany.
Kresse
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D002460:Cell Line; D058575:Decorin; D004187:Disaccharides / Q000032:analysis; D004705:Endocytosis / Q000187:drug effects*; D016326:Extracellular Matrix Proteins; D005944:Glucosamine / Q000031:analogs & derivatives; D006025:Glycosaminoglycans / Q000737:chemistry* / Q000494:pharmacology; D006493:Heparin / Q000737:chemistry / Q000494:pharmacology; D006497:Heparan Sulfate / Q000737:chemistry / Q000494:pharmacology; D009844:Oligosaccharides / Q000737:chemistry; D011133:Polysaccharide-Lyases / Q000378:metabolism; D011485:Protein Binding; D011509:Proteoglycans / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism*; D013431:Sulfates / Q000737:chemistry
D058575:Decorin; D004187:Disaccharides; D016326:Extracellular Matrix Proteins; D005944:Glucosamine; D006025:Glycosaminoglycans; D006493:Heparin; D006497:Heparan Sulfate; D009844:Oligosaccharides; D011133:Polysaccharide-Lyases; D011509:Proteoglycans; D011956:Receptors, Cell Surface; D013431:Sulfates; C021349:heparitinsulfate lyase