Heterogeneity of proteoglycans in monkey corneal stroma.
The proteoglycans of the cynomolgus monkey corneal stroma were isolated and characterized by using a combination of physiochemical and biochemical methods. Proteoglycans were biosynthetically radiolabeled by incubating whole corneas in medium containing [35S]sulfate and either [3H]serine or [3H]mannose as precursors. Macromolecules were extracted from the corneal stromas with 4 M guanidine-HCl. After dialysis into 8 M urea, proteoglycans in the extracts were initially purified by DEAE-cellulose chromatography. A portion of the proteoglycan fraction was digested with chondroitinase ABC, and the keratan sulfate proteoglycans were then isolated by rechromatography of the digest on DEAE-cellulose. Another portion of the proteoglycan fraction was digested with endo-beta-galactosidase and the dermatan sulfate-proteoglycans were then isolated by chromatography of the digest on Sepharose CL-4B. Each proteoglycan population was further fractionated by chromatography on concanavalin A-Sepharose and by CsCl density gradient centrifugation. Four subpopulations for both the keratan sulfate proteoglycans and the dermatan sulfate proteoglycans were isolated. Based on differences in binding to concanavalin A-Sepharose, buoyant densities, and glycosaminoglycan content, subpopulations of each proteoglycan differ by the number and properties of both the glycosaminoglycan chains and the mannose-containing oligosaccharides attached to their protein core.
D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002845:Chromatography / Q000379:methods; D003315:Cornea / Q000378:metabolism*; D003319:Corneal Stroma / Q000378:metabolism*; D003871:Dermatan Sulfate / Q000302:isolation & purification; D007632:Keratan Sulfate / Q000302:isolation & purification; D008252:Macaca fascicularis; D010206:Papain; D011509:Proteoglycans / Q000302:isolation & purification*
D011509:Proteoglycans; D003871:Dermatan Sulfate; D007632:Keratan Sulfate; D010206:Papain