Biochemical studies on the sulphated glycosaminoglycan fraction of skin fibroblasts cultured from a patient with the Hurler syndrome.
1. The metabolism of the sulphated glycosaminoglycan fraction in cultured skin fibroblasts derived from a patient with the Hurler syndrome and from a normal subject was studied. Two labelled precursors, Na(2) (35)SO(4) and d-[2-(3)H]glucose, were used and their intracellular fates during uptake and ;chase' periods were assessed after separation of sulphated glycosaminoglycans from hyaluronic acid. After 4 or 8h of exposure to culture medium containing both labels, [(35)S]sulphate incorporation into the sulphated glycosaminoglycan fraction was twofold greater in Hurler-syndrome cells than in normal cells. At the same time, the rate of incorporation of [(3)H]glucose into the sulphated glycosaminoglycan fraction was approximately the same for both cell types. Consequently, an increased (35)S/(3)H ratio (nmol of [(35)S]sulphate incorporated/nmol of [(3)H]glucose incorporated) was observed for Hurler-syndrome cells compared with normal cells. 2. The results of ;chase' experiments revealed that although the expected loss and relative retention of labelled sulphate occurred in the sulphated glycosaminoglycan fraction of normal and Hurler-syndrome cells, both cell types retained all of their radioactivity derived from [(3)H]glucose. 3. After 34h exposure to a ;corrective-factor' preparation from urine, the sulphated glycosaminoglycan content (as hexosamine and [(35)S]sulphate) of the Hurler-syndrome cells approached normal values. At the same time, there was an increase in specific radioactivity of ;corrected' Hurler-syndrome cells.
D002478:Cells, Cultured; D002852:Chromatography, Ion Exchange; D005347:Fibroblasts / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006025:Glycosaminoglycans / Q000378:metabolism* / Q000494:pharmacology; D006026:Glycoside Hydrolases / Q000652:urine; D006601:Hexoses; D006801:Humans; D007553:Isotope Labeling; D008059:Mucopolysaccharidosis I / Q000235:genetics* / Q000378:metabolism; D012867:Skin / Q000187:drug effects / Q000378:metabolism*; D013431:Sulfates / Q000378:metabolism; D013460:Sulfur Isotopes; D013997:Time Factors; D014316:Tritium; D014462:Ultrafiltration; D014574:Uronic Acids
D006025:Glycosaminoglycans; D006601:Hexoses; D013431:Sulfates; D013460:Sulfur Isotopes; D014574:Uronic Acids; D014316:Tritium; D006026:Glycoside Hydrolases; D005947:Glucose