title: Molecular attributes of bovine aortic endothelial cell heparan sulfate.
abstract: Heparan sulfate (HS) secreted into the medium of bovine aortic endothelial cell (BAEC) cultures was subjected to chemical and enzymatic degradation followed by analysis using gel-filtration and ion-exchange chromatography. Treatment with HNO2 showed that 41% of the disaccharides were N-sulfated. Degradation by Heparin lyases I (Hep I) showed that 8-9% of the disaccharides contained IdoA(2S) residues. Heparin lyase III (Hep III) degradation produced mainly disaccharides with 67% of the molecules glycosidic linkages susceptible to cleavage. Further degradation of Hep III-resistant fragments with Hep I showed that IdoA(2S) residues were predominantly positioned centrally within the repeating GlcNSO3(+/- 6S)alpha 1-4IdoA containing domains. Digestion with a mixture of Heparin lyases I, II and III degraded the molecule almost entirely to disaccharides, with small amounts of tetrasaccharides containing resistant linkages, suggesting the presence of 3-O sulfated GlcNSO3. Further analysis of the disaccharide products by ion-exchange chromatography and comparison with the data from single enzymatic digestion, allowed an estimate of the disaccharide composition to be made. The results suggest an ordered arrangement of structural domains; however, variations in the structure of these domains results in a heterogeneous population of HS chains. It is suggested that biosynthetic differences in HS structure may act as a regulator of bFGF induced cellular responses.
authors.affiliation: Department of Clinical Research, Christie Hospital, Manchester, United Kingdom.
authors.forename: D A
authors.initials: DA
authors.lastname: Pye
authors.forename: S
authors.initials: S
authors.lastname: Kumar
mesh_terms: D000818:Animals; D001011:Aorta; D002240:Carbohydrate Sequence; D002417:Cattle; D002478:Cells, Cultured; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004187:Disaccharides / Q000302:isolation & purification; D004730:Endothelium, Vascular / Q000737:chemistry*; D019763:Heparin Lyase; D006497:Heparan Sulfate / Q000737:chemistry* / Q000302:isolation & purification; D007067:Iduronic Acid / Q000032:analysis; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000302:isolation & purification; D011133:Polysaccharide-Lyases
chemical_list: D004187:Disaccharides; D019763:Heparin Lyase; D006497:Heparan Sulfate; D007067:Iduronic Acid; D009844:Oligosaccharides; D011133:Polysaccharide-Lyases