Molecular attributes of bovine aortic endothelial cell heparan sulfate.
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.
Pye
D A
DA
Department of Clinical Research, Christie Hospital, Manchester, United Kingdom.
Kumar
S
S
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
D004187:Disaccharides; D019763:Heparin Lyase; D006497:Heparan Sulfate; D007067:Iduronic Acid; D009844:Oligosaccharides; D011133:Polysaccharide-Lyases