Synthesis of glycosaminoglycans by cloned bovine endothelial cells cultured on collagen gels

D J Winterbourne, A M Schor, J T Gallagher

    Research output: Contribution to journalArticlepeer-review

    12 Citations (Scopus)

    Abstract

    Sulphated glycosaminoglycans have been analysed in cloned bovine aortic endothelial cells cultured on collagen gels after incubation with [3H]glucosamine and Na2(35)SO4. Radioactive products were analysed in the culture medium, in sequential collagenase and trypsin extracts of the cell monolayer and the associated extracellular matrix, and in the remaining viable cells. Heparan sulphate and chondroitin sulphate were found in each compartment: the heparan sulphate had a low degree of sulphation (approximately 0.4 N-sulphate and 0.2 O-sulphate groups per disaccharide unit on average). In the nitrous acid scission products of heparan sulphate, O-sulphated substituents were confined to disaccharide and tetrasaccharide fragments, indicating that local regions of the chain (which might be susceptible to excission by the platelet endoglycosidase) are highly sulphated. Only minor structural differences in heparan sulphate were observed between the various compartments. In contrast the chondroitin sulphate found in the collagenase extract had a higher iduronic acid content than corresponding material in the trypsin extract and the culture medium, indicating that collagenase and trypsin may extract glycosaminoglycans from different regions of the extracellular and pericellular matrix.

    Original languageEnglish
    Pages (from-to)271-7
    Number of pages7
    JournalEuropean Journal of Biochemistry
    Volume135
    Issue number2
    DOIs
    Publication statusPublished - Sept 1983

    Keywords

    • Animals
    • Cattle
    • Chemical Phenomena
    • Chemistry
    • Chondroitin Sulfates/isolation & purification
    • Chromatography/methods
    • Clone Cells
    • Collagen
    • Culture Media
    • Endothelium/metabolism
    • Gels
    • Glycosaminoglycans/biosynthesis
    • Heparitin Sulfate

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