Biosynthesis of dermatan sulphate. Assay and properties of the uronosyl C-5 epimerase.
During biosynthesis of dermatan sulphate D-glucuronate (GlcA) residues are converted to L-iduronate (IdoA) residues via the reaction [Formula: see text]. The reaction occurs on the polymer level and is catalysed by a C-5 uronosyl epimerase. The reversible release of the C-5 hydrogen was utilized as a measure of the enzyme activity with 5-3H-labelled chondroitin as a substrate. 3H released during incubation was distilled and quantified by liquid-scintillation counting. The epimerase has a low pH optimum (5.6) and requires divalent cations, Mn2+ being the most efficient for activity. The Km for chondroitin is 1.2 x 10(-4) M. The epimerase is largely associated with the microsomal fractions (90%). Two-thirds of the activity can be solubilized by detergents. Microsomes from cultured fibroblasts contain two different uronosyl epimerases, one for the biosynthesis of heparan sulphate and one for that of dermatan sulphate. The two epimerases have different cofactor and pH requirements.
D000818:Animals; D002238:Carbohydrate Epimerases / Q000378:metabolism*; D002413:Cations, Divalent / Q000494:pharmacology; D002417:Cattle; D002807:Chondroitin / Q000031:analogs & derivatives*; D003871:Dermatan Sulfate / Q000096:biosynthesis*; D006497:Heparan Sulfate / Q000378:metabolism; D066298:In Vitro Techniques; D007527:Isoenzymes / Q000378:metabolism; D010724:Phosphoadenosine Phosphosulfate / Q000494:pharmacology; D011134:Polysaccharides / Q000494:pharmacology; D013347:Subcellular Fractions / Q000201:enzymology
D002238:Carbohydrate Epimerases; D002413:Cations, Divalent; D002807:Chondroitin; D003871:Dermatan Sulfate; D006497:Heparan Sulfate; D007527:Isoenzymes; D010724:Phosphoadenosine Phosphosulfate; D011134:Polysaccharides; C019591:heparosan N-sulfate D-glucuronosyl 5-epimerase