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. 1974 Jun;139(3):565–581. doi: 10.1042/bj1390565

Hyaluronic acid in cartilage and proteoglycan aggregation

Timothy E Hardingham 1, Helen Muir 1
PMCID: PMC1166321  PMID: 4277512

Abstract

1. Dissociation of purified proteoglycan aggregates was shown to release an interacting component of buoyant density higher than that of the glycoprotein-link fraction of Hascall & Sajdera (1969). 2. This component, which produced an increase in hydrodynamic size of proteoglycans on gel chromatography, was isolated by ECTEOLA-cellulose ion-exchange chromatography and identified as hyaluronic acid. 3. The effect of pH of extraction showed that the proportion of proteoglycan aggregates isolated from cartilage was greatest at pH4.5. 4. The proportion of proteoglycans able to interact with hyaluronic acid decreased when extracted above or below pH4.5, whereas the amount of hyaluronic acid extracted appeared constant from pH3.0 to 8.5. 5. Sequential extraction of cartilage with 0.15m-NaCl at neutral pH followed by 4m-guanidinium chloride at pH4.5 was shown to yield predominantly non-aggregated and aggregated proteoglycans respectively. 6. Most of the hyaluronic acid in cartilage, representing about 0.7% of the total uronic acid, was associated with proteoglycan aggregates. 7. The non-aggregated proteoglycans were unable to interact with hyaluronic acid and were of smaller size, lower protein content and lower keratan sulphate content than the disaggregated proteoglycans. Together with differences in amino acid composition this suggested that each type of proteoglycan contained different protein cores.

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Selected References

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  1. 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]
  2. Baker J. R., Rodén L., Stoolmiller A. C. Biosynthesis of chondroitin sulfate proteoglycan. Xylosyl transfer to Smith-degraded cartilage proteoglycan and other exogenous acceptors. J Biol Chem. 1972 Jun 25;247(12):3838–3847. [PubMed] [Google Scholar]
  3. Brandt K. D., Muir H. Differences in composition and size of protein-polysaccharides extracted from pig articular cartilage. FEBS Lett. 1969 Jul;4(1):16–18. doi: 10.1016/0014-5793(69)80183-3. [DOI] [PubMed] [Google Scholar]
  4. Brandt K. D., Muir H. Heterogeneity of protein-polysaccharides of porcine articular cartilage. The sequential extraction of chondroitin sulphate-proteins with iso-osmotic neutral sodium acetate. Biochem J. 1971 Jan;121(2):261–270. doi: 10.1042/bj1210261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brandt K. D., Tsiganos C. P., Muir H. Immunological relationships between proteoglycans of different hydrodynamic size from articular cartilage of foetal and mature pigs. Biochim Biophys Acta. 1973 Sep 14;320(2):453–468. doi: 10.1016/0304-4165(73)90326-7. [DOI] [PubMed] [Google Scholar]
  6. DODGSON K. S. Determination of inorganic sulphate in studies on the enzymic and non-enzymic hydrolysis of carbohydrate and other sulphate esters. Biochem J. 1961 Feb;78:312–319. doi: 10.1042/bj0780312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gregory J. D. Multiple aggregation factors in cartilage proteoglycan. Biochem J. 1973 Jun;133(2):383–386. doi: 10.1042/bj1330383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hardingham T. E., Muir H. Binding of oligosaccharides of hyaluronic acid to proteoglycans. Biochem J. 1973 Dec;135(4):905–908. doi: 10.1042/bj1350905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hardingham T. E., Muir H. Biosynthesis of proteoglycans in cartilage slices. Fractionation by gel chromatography and equilibrium density-gradient centrifugation. Biochem J. 1972 Feb;126(4):791–803. doi: 10.1042/bj1260791. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hardingham T. E., Muir H. The effect of temperature on the biosynthesis of chondroitin 4-sulphate in cartilage slices in vitro. FEBS Lett. 1970 Sep 6;9(3):145–148. doi: 10.1016/0014-5793(70)80339-8. [DOI] [PubMed] [Google Scholar]
  11. Hardingham T. E., Muir H. The specific interaction of hyaluronic acid with cartillage proteoglycans. Biochim Biophys Acta. 1972 Sep 15;279(2):401–405. doi: 10.1016/0304-4165(72)90160-2. [DOI] [PubMed] [Google Scholar]
  12. Hascall V. C., Riolo R. L. Characteristics of the protein-keratan sulfate core and of keratan sulfate prepared from bovine nasal cartilage proteoglycan. J Biol Chem. 1972 Jul 25;247(14):4529–4538. [PubMed] [Google Scholar]
  13. Hascall V. C., Sajdera S. W. Physical properties and polydispersity of proteoglycan from bovine nasal cartilage. J Biol Chem. 1970 Oct 10;245(19):4920–4930. [PubMed] [Google Scholar]
  14. Hascall V. C., Sajdera S. W. Proteinpolysaccharide complex from bovine nasal cartilage. The function of glycoprotein in the formation of aggregates. J Biol Chem. 1969 May 10;244(9):2384–2396. [PubMed] [Google Scholar]
  15. Heinegård D. Extraction, fractionation and characterization of proteoglycans from bovine tracheal cartilage. Biochim Biophys Acta. 1972 Nov 28;285(1):181–192. doi: 10.1016/0005-2795(72)90190-0. [DOI] [PubMed] [Google Scholar]
  16. Heinegård D., Gardell S. Studies on protein-polysaccharide complex (proteoglycan) from human nucleus pulposus. I. Isolation and preliminary characterisation. Biochim Biophys Acta. 1967 Oct 9;148(1):164–171. doi: 10.1016/0304-4165(67)90292-9. [DOI] [PubMed] [Google Scholar]
  17. Heinegård D. Hyaluronidase digestion and alkaline treatment of bovine tracheal cartilage proteoglycans. Isolation and characterisation of different keratan sulfate proteins. Biochim Biophys Acta. 1972 Nov 28;285(1):193–207. doi: 10.1016/0005-2795(72)90191-2. [DOI] [PubMed] [Google Scholar]
  18. KIMMEL J. R., SMITH E. L. Crystalline papain. I. Preparation, specificity, and activation. J Biol Chem. 1954 Apr;207(2):515–531. [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. MATHEWS M. B., LOZAITYTE I. Sodium chondroitin sulfate-protein complexes of cartilage. I. Molecular weight and shape. Arch Biochem Biophys. 1958 Mar;74(1):158–174. doi: 10.1016/0003-9861(58)90210-8. [DOI] [PubMed] [Google Scholar]
  21. MUIR H. The nature of the link between protein and carbohydrate of a chondroitin sulphate complex from hyaline cartilage. Biochem J. 1958 Jun;69(2):195–204. doi: 10.1042/bj0690195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mashburn T. A., Jr, Hoffman P. Comparative fractionation studies of cartilage proteinpolysaccharides. J Biol Chem. 1971 Nov;246(21):6497–6506. [PubMed] [Google Scholar]
  23. Mathews M. B. Comparative biochemistry of chondroitin sulphate-proteins of cartilage and notochord. Biochem J. 1971 Nov;125(1):37–46. doi: 10.1042/bj1250037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mayes R. W., Mason R. M., Griffin D. C. The composition of cartilage proteoglycans. An investigation using high- and low-inonic-strength extraction procedures. Biochem J. 1973 Mar;131(3):541–553. doi: 10.1042/bj1310541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Muir H. The structure and metabolism of mucopolysaccharides (glycosaminoglycans) and the problem of the mucopolysaccharidoses. Am J Med. 1969 Nov;47(5):673–690. doi: 10.1016/0002-9343(69)90163-6. [DOI] [PubMed] [Google Scholar]
  26. PARTRIDGE S. M., DAVIS H. F., ADAIR G. S. The chemistry of connective tissues. 6. The constitution of the chondroitin sulphate-protein complex in cartilage. Biochem J. 1961 Apr;79:15–26. doi: 10.1042/bj0790015. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rosenberg L., Pal S., Beale R., Schubert M. A comparison of proteinpolysaccharides of bovine nasal cartilage isolated and fractionated by different methods. J Biol Chem. 1970 Aug 25;245(16):4112–4122. [PubMed] [Google Scholar]
  28. Sajdera S. W., Hascall V. C. Proteinpolysaccharide complex from bovine nasal cartilage. A comparison of low and high shear extraction procedures. J Biol Chem. 1969 Jan 10;244(1):77–87. [PubMed] [Google Scholar]
  29. Simůnek Z., Muir H. Changes in the protein-polysaccharides of pig articular cartilage during prenatal life, development and old age. Biochem J. 1972 Feb;126(3):515–523. doi: 10.1042/bj1260515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tsiganos C. P., Hardingham T. E., Muir H. Proteoglycans of cartilage: an assessment of their structure. Biochim Biophys Acta. 1971 Feb 16;229(2):529–534. doi: 10.1016/0005-2795(71)90216-9. [DOI] [PubMed] [Google Scholar]
  31. Tsiganos C. P., Muir H. Studies on protein-polysaccharides from pig laryngeal cartilage. Extraction and purification. Biochem J. 1969 Aug;113(5):879–884. doi: 10.1042/bj1130879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tsiganos C. P., Muir H. Studies on protein-polysaccharides from pig laryngeal cartilage. Heterogeneity, fractionation and characterization. Biochem J. 1969 Aug;113(5):885–894. doi: 10.1042/bj1130885. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. WOESSNER J. F., Jr The determination of hydroxyproline in tissue and protein samples containing small proportions of this imino acid. Arch Biochem Biophys. 1961 May;93:440–447. doi: 10.1016/0003-9861(61)90291-0. [DOI] [PubMed] [Google Scholar]

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