title: Molecular organization and antiproliferative domains of arterial tissue heparan sulfate.
abstract: Heparan sulfate isolated from mammalian arterial tissue inhibits the growth of homologous arterial smooth muscle cells when added to subconfluent cell cultures at a concentration of 50 to 100 micrograms/ml culture medium. Disintegration of the heparan sulfate molecule by hydrazinolysis that deacetylates N-acetylglucosaminyl residues and by subsequent treatment with nitrous acid at pH 3.9 results in the formation of a mixture of oligosaccharides which was further resolved into sulfate-enriched oligosaccharides with antiproliferative activity in an in vitro bioassay system. A decasaccharide and dodeca/tetradecasaccharide fraction had a significantly higher antiproliferative effect on arterial smooth muscle cells than the native heparan sulfate molecule. The antiproliferative oligosaccharides have a sulfate content of 0.9 to 1.2 sulfate groups/disaccharide unit and consist of 60 to 70% monosulfated, disulfated, and trisulfated disaccharide units. Up to 32% of the sulfate groups were in 2-position of the uronic acid. In contrast, nitrous acid degradation of heparan sulfate at pH 1.5, which cleaves glycosidic linkages of N-sulfoglucosaminyl residues, results in the formation of sulfate-poor or sulfate-free oligosaccharides without antiproliferative potency. The results indicate that (a) heparan sulfate has a heterogeneous molecular organization where sulfate-rich domains are separated by sulfate-poor sequences and that (b) the antiproliferative activity of heparan sulfate resides in domains enriched with 2-O-sulfated uronic acid residues.
authors.affiliation: Institut für Arterioskleroseforschung an der Universität Münster, Deutschland.
authors.forename: A
authors.initials: A
authors.lastname: Schmidt
authors.forename: G
authors.initials: G
authors.lastname: Lemming
authors.forename: K
authors.initials: K
authors.lastname: Yoshida
authors.forename: E
authors.initials: E
authors.lastname: Buddecke
mesh_terms: D000818:Animals; D001011:Aorta / Q000737:chemistry; D002417:Cattle; D002455:Cell Division / Q000502:physiology; D002478:Cells, Cultured; D004187:Disaccharides / Q000032:analysis; D006497:Heparan Sulfate / Q000737:chemistry*; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000737:chemistry*; D009608:Nitrous Acid; D009844:Oligosaccharides / Q000302:isolation & purification; D011506:Proteins / Q000737:chemistry; D012685:Sepharose; D013431:Sulfates / Q000032:analysis; D014574:Uronic Acids / Q000032:analysis
chemical_list: D004187:Disaccharides; D006497:Heparan Sulfate; D009608:Nitrous Acid; D009844:Oligosaccharides; D011506:Proteins; D012685:Sepharose; D013431:Sulfates; D014574:Uronic Acids