Mechanism for the decreased biosynthesis of cartilage proteoglycan in the scorbutic guinea pig.
Our previous work showed that vitamin C deficiency caused about a 70-80% decrease in the incorporation of [35S]sulfate into proteoglycan of guinea pig costal cartilage, coordinately with a decrease in collagen synthesis (Bird, T. A., Spanheimer, R. G., and Peterkofsky, B. (1986) Arch. Biochem. Biophys. 246, 42-51). We examined the mechanism for decreased proteoglycan synthesis by labeling normal and scorbutic cartilage in vitro with radioactive precursors. Proteoglycan monomers from scorbutic tissue were of a slightly smaller average hydrodynamic size than normal but there was no difference in the size of the glycosaminoglycan chains isolated after papain digestion. The type of glycosaminoglycans synthesized and the degree of sulfation were unaffected as determined by chondroitinase ABC digestion and duel labeling with [35S]sulfate and [3H]glucosamine. Conversion of [3H]glucosamine to [3H]galactosamine also was unimpaired. There was about a 40% decrease in core protein synthesis, measured by [14C]serine incorporation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Nevertheless, decreased incorporation of [35S]sulfate into scorbutic tissue persisted in the presence of p-nitrophenyl-beta-D-xyloside and cycloheximide, which indicated that the site of the scorbutic defect was beyond core protein synthesis and xylosylation. Galactosyltransferase activity in scorbutic cartilage decreased to about one-third the levels in control samples in parallel with the decreases in proteoglycan and collagen synthesis. Our results suggest that the step catalyzed by this enzyme activity, the addition of galactose to xylose prior to chondroitin sulfate chain elongation, is the major site of the scorbutic defect in proteoglycan synthesis. Decreased enzyme activity may be related to increased cortisol levels in scorbutic serum.
D000818:Animals; D001206:Ascorbic Acid Deficiency / Q000378:metabolism*; D002356:Cartilage / Q000378:metabolism*; D002808:Chondroitin Lyases / Q000378:metabolism; D002850:Chromatography, Gel; D003513:Cycloheximide / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005688:Galactosamine / Q000378:metabolism; D005944:Glucosamine / Q000378:metabolism; D006027:Glycosides / Q000494:pharmacology; D006168:Guinea Pigs; D006854:Hydrocortisone / Q000097:blood; D011509:Proteoglycans / Q000096:biosynthesis*; D013431:Sulfates / Q000378:metabolism
D006027:Glycosides; D011509:Proteoglycans; D013431:Sulfates; C010807:xylosides; D005688:Galactosamine; D003513:Cycloheximide; D002808:Chondroitin Lyases; D005944:Glucosamine; D006854:Hydrocortisone