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. 1991 Jun 1;276(Pt 2):533–539. doi: 10.1042/bj2760533

Biosynthesis of dermatan sulphate proteoglycans. The effect of beta-D-xyloside addition on the polymer-modification process in fibroblast cultures.

L Cöster 1, J Hernnäs 1, A Malmström 1
PMCID: PMC1151124  PMID: 2049079

Abstract

Incubation of cultured fibroblasts with p-nitrophenyl beta-D-xyloside resulted in a concentration-dependent increase in galactosaminoglycan synthesis. At low concentration of added xyloside large and small radiolabelled proteoglycans and xyloside-bound polysaccharides were recovered from the medium, whereas at high concentrations only xyloside-bound polysaccharides were found. In the cell layer proteoglycans and xyloside-bound polysaccharides were found at all concentrations tested. Only galactosaminoglycan chains were polymerized on the xyloside primer. At low concentrations of added xyloside the structure of the galactosaminoglycans formed on the xyloside was similar to that of the small dermatan sulphate proteoglycan, i.e. mainly composed of L-iduronic acid-containing 4-sulphated disaccharides. With increasing concentration of added xyloside the co-polymeric structure of the small dermatan sulphate proteoglycan and the xyloside-bound polysaccharide was changed to contain a larger proportion of D-glucuronosyl residues with only slight changes in the sulphation pattern. No structural change in the polysaccharide chains of the large glucuronic acid-rich proteoglycans occurred. At 1 mM-xyloside, where no proteoglycans were formed, the polysaccharide was shorter and composed mainly of D-glucuronosyl-containing disaccharides with a ratio of 4-sulphate to 6-sulphate substituents of 1:2. This is similar to the structure of the large glucuronic acid-rich proteoglycan synthesized by these cells. Thus the main difference induced by the xyloside treatment was changed polymer modification at high xyloside concentrations. The specific activities of the polymer-modifying enzymes, uronosyl C-5-epimerase and 4-sulphotransferase, were therefore measured and found to be decreased by 30-50% in fibroblasts treated with high xyloside concentrations. It is suggested that the protein core is of importance for regulating the activity of the polymer-modifying enzymes.

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Selected References

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