title: The copolymeric structure of dermatan sulphate produced by cultured human fibroblasts. Different distribution of iduronic acid and glucuronic acid-containing units in soluble and cell-associated glycans.
abstract: The structure of dermatan [35S]sulphate-chondroitin [35S]sulphate copolymers synthesized and secreted by fibroblasts in culture was studied. 35S-labelled glycosaminoglycans were isolated from the medium, a trypsin digest of the cells and the cell residue after 72h of 35SO42-incorporation. The galactosaminoglycan component (dermatan sulphatechondroitin sulphate copolymers) was isolated and subjected to various degradation procedures including digestion with testicular hyaluronidase, chondroitinase-AC and-ABC and periodate oxidation followed by alkaline elimination. The galactosaminoglycans from the various sources displayed significant structural differences with regard to the distribution of various repeating units, i.e. IdUA-GalNAc-SO4 (L-iduronic acid-N-acetyl-galactosamine sulphate), GlcUA-GalNAc-SO4 (D-glucuronic acid-N-acetylgalactosamine-sulphate) and IdUA(-SO4)-GalNAc (L-iduronosulphate-N-acetylgalactosamine). The galactosaminoglycans of the cell residue contained larger amounts of IdUA-GalNAc-SO4 than did those isolated from the medium or those released by trypsin. In contrast, the glycans from the latter 2 sources contained large proportions of periodate-resistant repeat periods [GlcUA-GalNAc-SO4 and IdUA(-SO4)-GalNAc]. Periods containing L-iduronic acid sulphate were particularly prominent in copolymers found in the medium. Kinetic studies indicated that the 35S-labelled glycosaminoglycan of the cell residue accumulated radioactivity more slowly than did the glycans of other fractions, indicating that the material remaining with the cells was not exclusively a precursor of the secreted polymers. The presence of copolymers rich in glucuronic acid or iduronic acid sulphate residues in the soluble fractions may be the result of selective secretion from the cells. Alternatively, extracellular, polymer-level modifications such as C-5 inversion of L-iduronic acid to D-glucuronic acid, or sulphate rearrangements, would yield similar results.
mesh_terms: D002478:Cells, Cultured; D002807:Chondroitin / Q000031:analogs & derivatives*; D002848:Chromatography, DEAE-Cellulose; D003871:Dermatan Sulfate / Q000032:analysis / Q000096:biosynthesis; D004590:Electrophoresis, Paper; D005965:Glucuronates / Q000032:analysis; D006025:Glycosaminoglycans / Q000378:metabolism; D007067:Iduronic Acid / Q000032:analysis; D008968:Molecular Conformation; D011134:Polysaccharides / Q000032:analysis; D013462:Sulfur Radioisotopes; D013997:Time Factors
chemical_list: D005965:Glucuronates; D006025:Glycosaminoglycans; D011134:Polysaccharides; D013462:Sulfur Radioisotopes; D003871:Dermatan Sulfate; D007067:Iduronic Acid; D002807:Chondroitin