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. 1972 Jun;128(2):205–213. doi: 10.1042/bj1280205

The biosynthesis in vitro of keratan sulphate in bovine cornea

C J Handley 1, C F Phelps 1
PMCID: PMC1173756  PMID: 4263792

Abstract

1. Bovine corneas were incubated in vitro with [U-14C]glucose. 2. The glycosaminoglycans of corneal stroma were isolated and fractionated on cetylpyridinium chloride–cellulose columns. The major components were keratan sulphate (71%), chondroitin 4-sulphate (17%) and chondroitin 6-sulphate (4%). 3. The acid-soluble nucleotides and intermediates of glycosaminoglycan biosynthesis of corneal stroma were separated on Dowex 1 (formate form) and the tissue content and cellular concentrations were determined. 4. The rates of synthesis of the intermediates of glycosaminoglycan biosynthesis in corneal stroma were determined. 5. The incorporation of radioactivity from [U-14C]glucose into the uronic acid and hexosamine components of the glycosaminoglycans present in corneal stroma were measured and the turnover rates of these polymers were calculated. It was found that keratan sulphate was turning over in about 723h and chondroitin 6-sulphate in 251h.

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Selected References

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  1. ANSETH A., LAURENT T. C. Studies on corneal polysaccharides. I. Separation. Exp Eye Res. 1961 Sep;1:25–38. doi: 10.1016/s0014-4835(61)80005-5. [DOI] [PubMed] [Google Scholar]
  2. ANSETH A. Studies on corneal polysaccharides. III. Topographic and comparative biochemistry. Exp Eye Res. 1961 Dec;1:106–115. doi: 10.1016/s0014-4835(61)80015-8. [DOI] [PubMed] [Google Scholar]
  3. ANTONOPOULOS C. A., BORELIUS E., GARDELL S., HAMNSTROM B., SCOTT J. E. The precipitation of polyanions by long-chain aliphatic ammonium compounds. IV. Elution in salt solutions of mucopolysaccharide-quaternary ammonium complexes adsorbed on a support. Biochim Biophys Acta. 1961 Dec 9;54:213–226. doi: 10.1016/0006-3002(61)90360-2. [DOI] [PubMed] [Google Scholar]
  4. BITTER T., MUIR H. M. A modified uronic acid carbazole reaction. Anal Biochem. 1962 Oct;4:330–334. doi: 10.1016/0003-2697(62)90095-7. [DOI] [PubMed] [Google Scholar]
  5. DAVIDSON E. A., SMALL W. Metabolism in vivo of connective-tissue mucopolysaccharides. I. Chondroitin sulfate C and keratosulfate of nucleus pulposus. Biochim Biophys Acta. 1963 Mar 5;69:445–452. doi: 10.1016/0006-3002(63)91292-7. [DOI] [PubMed] [Google Scholar]
  6. Fransson L. A., Anseth A. Studies on corneal polysaccharides. IV. Chromatography of corneal glycosaminoglycans on ECTEOLA cellulose using formate buffers as eluting solvents. Exp Eye Res. 1967 Apr;6(2):107–119. doi: 10.1016/s0014-4835(67)80061-7. [DOI] [PubMed] [Google Scholar]
  7. GLASER L. The biosynthesis of N-acetylgalactosamine. J Biol Chem. 1959 Nov;234:2801–2805. [PubMed] [Google Scholar]
  8. Gainey P. A., Phelps C. F. Uridine diphosphate glucuronic acid production and utilization in various tissues actively synthesizing glycosaminoglycans. Biochem J. 1972 Jun;128(2):215–227. doi: 10.1042/bj1280215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. HURLBERT R. B., SCHMITZ H., BRUMM A. F., POTTER V. R. Nucleotide metabolism. II. Chromatographic separation of acid-soluble nucleotides. J Biol Chem. 1954 Jul;209(1):23–39. [PubMed] [Google Scholar]
  10. Hardingham T. E., Phelps C. F. Studies on the incorporation of (U-14C)glucose and (35S)sulphate into the acid glycosaminoglycans of neonatal rat skin. Biochem J. 1970 Oct;119(5):885–893. doi: 10.1042/bj1190885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hardingham T. E., Phelps C. F. The glycosaminoglycans of neonatal rat skin. Biochem J. 1970 May;117(5):813–818. doi: 10.1042/bj1170813. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hardingham T. E., Phelps C. F. The tissue content and turnover rates of intermediates in the biosynthesis of glycosaminoglycans in young rat skin. Biochem J. 1968 Jun;108(1):9–16. doi: 10.1042/bj1080009. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. KORNFELD S., KORNFELD R., NEUFELD E. F., O'BRIEN P. J. THE FEEDBACK CONTROL OF SUGAR NUCLEOTIDE BIOSYNTHESIS IN LIVER. Proc Natl Acad Sci U S A. 1964 Aug;52:371–379. doi: 10.1073/pnas.52.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LELOIR L. F. The enzymatic transformation of uridine diphosphate glucose into a galactose derivative. Arch Biochem Biophys. 1951 Sep;33(2):186–190. doi: 10.1016/0003-9861(51)90096-3. [DOI] [PubMed] [Google Scholar]
  15. MEYER K., LINKER A., DAVIDSON E. A., WEISSMANN B. The mucopolysaccharides of bovine cornea. J Biol Chem. 1953 Dec;205(2):611–616. [PubMed] [Google Scholar]
  16. NEUFELD E. F., HALL C. W. INHIBITION OF UDP-D-GLUCOSE DEHYDROGENASE BY UDP-D-XYLOSE: A POSSIBLE REGULATORY MECHANISM. Biochem Biophys Res Commun. 1965 May 3;19:456–461. doi: 10.1016/0006-291x(65)90146-4. [DOI] [PubMed] [Google Scholar]
  17. Randerath E., Randerath K. Ion-exchange thin-layer chromatography. XII. Quantitative elution and microdetermination of nucleoside monophosphates, ATP, and other nucleotide coenzymes. Anal Biochem. 1965 Jul;12(1):83–93. doi: 10.1016/0003-2697(65)90145-4. [DOI] [PubMed] [Google Scholar]
  18. SCHILLER S., MATHEWS M. B., CIFONELLI J. A., DORFMAN A. The metabolism of mucopolysaccharides in animals. III. Further studies on skin utilizing C14-glucose, C14-acetate, and S35-sodium sulfate. J Biol Chem. 1956 Jan;218(1):139–145. [PubMed] [Google Scholar]
  19. WACHTL C., KINSEY V. E. Studies on the crystalline lens. VIII. A synthetic medium for lens culture and the effects of various constituents on cell division in the epithelium. Am J Ophthalmol. 1958 Nov;46(5 Pt 2):288–293. [PubMed] [Google Scholar]
  20. Winterburn P. J., Phelps C. F. Studies on the control of hexosamine biosynthesis by glucosamine synthetase. Biochem J. 1971 Feb;121(4):711–720. doi: 10.1042/bj1210711. [DOI] [PMC free article] [PubMed] [Google Scholar]

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