Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1986 Sep 1;238(2):571–580. doi: 10.1042/bj2380571

Articular cartilage cultured with interleukin 1. Increased release of link protein, hyaluronate-binding region and other proteoglycan fragments.

A Ratcliffe, J A Tyler, T E Hardingham
PMCID: PMC1147171  PMID: 3492199

Abstract

Pig articular cartilage was maintained in culture for 3 days with and without porcine interleukin 1. The proteoglycans remaining in the cartilage and those released into the medium were analysed by using radioimmunoassays for the hyaluronate-binding region, link protein and keratan sulphate. In interleukin 1-treated cultures after 3 days there was 38% release of total glycosaminoglycans into the medium, 18% release of binding region, 14% release of link protein and 20% release of keratan sulphate epitope, whereas in control cultures the proportions released were much less (16, 9, 10 and 7% respectively). Characterization of the proteoglycans in the media after 1.5 days and 3 days of culture showed that interleukin 1 promoted the release of proteoglycan of large average size and also the release of link protein and of low-Mr binding region which was unattached to proteoglycan. Both the link protein and binding region released were able to bind to exogenously added hyaluronate, whereas the proteoglycan in the medium was not. The proteoglycans extracted from cultured cartilage were similar to those from fresh cartilage: they contained a high proportion of aggregating proteoglycans and some low-Mr binding region. The proportion of this binding region extracted from the interleukin 1-treated cartilage was increased. The presence of interleukin 1 in the cultures therefore appeared to increase the rate of proteolytic degradation of proteoglycan in the matrix and to lead to a more rapid loss of intact binding region, of link protein and of large proteoglycan fragments into the medium.

Full text

PDF
575

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baracos V., Rodemann H. P., Dinarello C. A., Goldberg A. L. Stimulation of muscle protein degradation and prostaglandin E2 release by leukocytic pyrogen (interleukin-1). A mechanism for the increased degradation of muscle proteins during fever. N Engl J Med. 1983 Mar 10;308(10):553–558. doi: 10.1056/NEJM198303103081002. [DOI] [PubMed] [Google Scholar]
  2. Bayliss M. T., Ridgway G. D., Ali S. Y. Delayed aggregation of proteoglycans in adult human articular cartilage. Biosci Rep. 1984 Oct;4(10):827–833. doi: 10.1007/BF01138164. [DOI] [PubMed] [Google Scholar]
  3. Bayliss M. T., Ridgway G. D., Ali S. Y. Differences in the rates of aggregation of proteoglycans from human articular cartilage and chondrosarcoma. Biochem J. 1983 Dec 1;215(3):705–708. doi: 10.1042/bj2150705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Benya P. D., Nimni M. E. The stability of the collagen phenotype during stimulated collagen, glycosaminoglycan, and DNA synthesis by articular cartilage organ cultures. Arch Biochem Biophys. 1979 Feb;192(2):327–335. doi: 10.1016/0003-9861(79)90100-0. [DOI] [PubMed] [Google Scholar]
  5. Bonnet F., Dunham D. G., Hardingham T. E. Structure and interactions of cartilage proteoglycan binding region and link protein. Biochem J. 1985 May 15;228(1):77–85. doi: 10.1042/bj2280077. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  7. Dingle J. T. Heberden oration 1978. Recent studies on the control of joint damage: the contribution of the Strangeways Research Laboratory. Ann Rheum Dis. 1979 Jun;38(3):201–214. doi: 10.1136/ard.38.3.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dingle J. T., Saklatvala J., Hembry R., Tyler J., Fell H. B., Jubb R. A cartilage catabolic factor from synovium. Biochem J. 1979 Oct 15;184(1):177–180. doi: 10.1042/bj1840177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fairbanks G., Steck T. L., Wallach D. F. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry. 1971 Jun 22;10(13):2606–2617. doi: 10.1021/bi00789a030. [DOI] [PubMed] [Google Scholar]
  10. Farndale R. W., Sayers C. A., Barrett A. J. A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. Connect Tissue Res. 1982;9(4):247–248. doi: 10.3109/03008208209160269. [DOI] [PubMed] [Google Scholar]
  11. Fell H. B., Jubb R. W. The effect of synovial tissue on the breakdown of articular cartilage in organ culture. Arthritis Rheum. 1977 Sep-Oct;20(7):1359–1371. doi: 10.1002/art.1780200710. [DOI] [PubMed] [Google Scholar]
  12. Gowen M., Wood D. D., Ihrie E. J., Meats J. E., Russell R. G. Stimulation by human interleukin 1 of cartilage breakdown and production of collagenase and proteoglycanase by human chondrocytes but not by human osteoblasts in vitro. Biochim Biophys Acta. 1984 Feb 14;797(2):186–193. doi: 10.1016/0304-4165(84)90121-1. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Hardingham T. E. The role of link-protein in the structure of cartilage proteoglycan aggregates. Biochem J. 1979 Jan 1;177(1):237–247. doi: 10.1042/bj1770237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Hardingham T. Proteoglycans: their structure, interactions and molecular organization in cartilage. Biochem Soc Trans. 1981 Dec;9(6):489–497. doi: 10.1042/bst0090489. [DOI] [PubMed] [Google Scholar]
  16. Hascall V. C., Handley C. J., McQuillan D. J., Hascall G. K., Robinson H. C., Lowther D. A. The effect of serum on biosynthesis of proteoglycans by bovine articular cartilage in culture. Arch Biochem Biophys. 1983 Jul 1;224(1):206–223. doi: 10.1016/0003-9861(83)90205-9. [DOI] [PubMed] [Google Scholar]
  17. Hascall V. C. Interaction of cartilage proteoglycans with hyaluronic acid. J Supramol Struct. 1977;7(1):101–120. doi: 10.1002/jss.400070110. [DOI] [PubMed] [Google Scholar]
  18. Heath J. K., Saklatvala J., Meikle M. C., Atkinson S. J., Reynolds J. J. Pig interleukin 1 (catabolin) is a potent stimulator of bone resorption in vitro. Calcif Tissue Int. 1985 Jan;37(1):95–97. doi: 10.1007/BF02557686. [DOI] [PubMed] [Google Scholar]
  19. Heinegård D. Polydispersity of cartilage proteoglycans. Structural variations with size and buoyant density of the molecules. J Biol Chem. 1977 Mar 25;252(6):1980–1989. [PubMed] [Google Scholar]
  20. Heinegård D., Wieslander J., Sheehan J., Paulsson M., Sommarin Y. Separation and characterization of two populations of aggregating proteoglycans from cartilage. Biochem J. 1985 Jan 1;225(1):95–106. doi: 10.1042/bj2250095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Jubb R. W., Fell H. B. The breakdown of collagen by chondrocytes. J Pathol. 1980 Mar;130(3):159–167. doi: 10.1002/path.1711300304. [DOI] [PubMed] [Google Scholar]
  22. Mizel S. B., Dayer J. M., Krane S. M., Mergenhagen S. E. Stimulation of rheumatoid synovial cell collagenase and prostaglandin production by partially purified lymphocyte-activating factor (interleukin 1). Proc Natl Acad Sci U S A. 1981 Apr;78(4):2474–2477. doi: 10.1073/pnas.78.4.2474. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Morales T. I., Wahl L. M., Hascall V. C. The effect of bacterial lipopolysaccharides on the biosynthesis and release of proteoglycans from calf articular cartilage cultures. J Biol Chem. 1984 Jun 10;259(11):6720–6729. [PubMed] [Google Scholar]
  24. Ratcliffe A., Fryer P. R., Hardingham T. E. The distribution of aggregating proteoglycans in articular cartilage: comparison of quantitative immunoelectron microscopy with radioimmunoassay and biochemical analysis. J Histochem Cytochem. 1984 Feb;32(2):193–201. doi: 10.1177/32.2.6363519. [DOI] [PubMed] [Google Scholar]
  25. Ratcliffe A., Hardingham T. Cartilage proteoglycan binding region and link protein. Radioimmunoassays and the detection of masked determinants in aggregates. Biochem J. 1983 Aug 1;213(2):371–378. doi: 10.1042/bj2130371. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Saklatvala J., Curry V. A., Sarsfield S. J. Purification to homogeneity of pig leucocyte catabolin, a protein that causes cartilage resorption in vitro. Biochem J. 1983 Nov 1;215(2):385–392. doi: 10.1042/bj2150385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Saklatvala J., Pilsworth L. M., Sarsfield S. J., Gavrilovic J., Heath J. K. Pig catabolin is a form of interleukin 1. Cartilage and bone resorb, fibroblasts make prostaglandin and collagenase, and thymocyte proliferation is augmented in response to one protein. Biochem J. 1984 Dec 1;224(2):461–466. doi: 10.1042/bj2240461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Saklatvala J., Sarsfield S. J., Townsend Y. Pig interleukin 1. Purification of two immunologically different leukocyte proteins that cause cartilage resorption, lymphocyte activation, and fever. J Exp Med. 1985 Oct 1;162(4):1208–1222. doi: 10.1084/jem.162.4.1208. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Schwartz N. B., Habib G., Campbell S., D'Elvlyn D., Gartner M., Krueger R., Olson C., Philipson L. Synthesis and structure of proteoglycan core protein. Fed Proc. 1985 Feb;44(2):369–372. [PubMed] [Google Scholar]
  30. Tyler J. A. Articular cartilage cultured with catabolin (pig interleukin 1) synthesizes a decreased number of normal proteoglycan molecules. Biochem J. 1985 May 1;227(3):869–878. doi: 10.1042/bj2270869. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Tyler J. A. Chondrocyte-mediated depletion of articular cartilage proteoglycans in vitro. Biochem J. 1985 Jan 15;225(2):493–507. doi: 10.1042/bj2250493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Tyler J. A., Fell H. B., Lawrence C. E. The effect of cortisol on porcine articular tissues in organ culture. J Pathol. 1982 Aug;137(4):335–351. doi: 10.1002/path.1711370408. [DOI] [PubMed] [Google Scholar]
  33. Wiedemann H., Paulsson M., Timpl R., Engel J., Heinegård D. Domain structure of cartilage proteoglycans revealed by rotary shadowing of intact and fragmented molecules. Biochem J. 1984 Nov 15;224(1):331–333. doi: 10.1042/bj2240331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zanetti M., Ratcliffe A., Watt F. M. Two subpopulations of differentiated chondrocytes identified with a monoclonal antibody to keratan sulfate. J Cell Biol. 1985 Jul;101(1):53–59. doi: 10.1083/jcb.101.1.53. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES