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Biochem J. 1998 Oct 1; 335(Pt 1): 59–66.
PMCID: PMC1219752

Chondrocyte-mediated catabolism of aggrecan: aggrecanase-dependent cleavage induced by interleukin-1 or retinoic acid can be inhibited by glucosamine.


A rat chondrosarcoma cell line and bovine cartilage explants have been used to study the control of aggrecan degradation by chondrocytes treated with interleukin-1 (IL-1) or retinoic acid (RA). Aggrecan fragment analysis with anti-neo-epitope antibodies suggests that aggrecanase (an as yet unidentified enzyme) is the only aggrecan-degrading proteinase active in these cultures. With rat cells, aggrecanase converts the aggrecan core protein into two major G1-domain-bearing products (60 kDa with a C-terminal Glu-373, and 220 kDa with a C-terminal Glu-1459). Both products were quantified on a standardized Western analysis system with a G1-specific antibody. Immunoblots were analysed by scanning densitometry and the sensitivity, linearity and reproducibility of the assay were established. With rat cells the aggrecanase response to IL-1 was optimal at about 2 mM glutamine, but was progressively inhibited at higher concentrations, with about 90% inhibition at 10 mM glutamine. Such inhibition by glutamine was not, however, observed with bovine explants. On the other hand, marked inhibition of aggrecanase-dependent cleavage was observed with both rat cells and bovine explants when d(+)-glucosamine was included at concentrations above 2 mM. Inhibition was apparently not due to cytotoxicity or interference with IL-1 signalling, since biosynthetic activity was not inhibited and inhibition of the aggrecanase response was also obtained when RA was used as the catabolic stimulator. Possible mechanisms for the inhibition of the aggrecanase response by glucosamine in chondrocytes treated with IL-1 or RA are discussed.

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Selected References

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  • Luyten FP, Hascall VC, Nissley SP, Morales TI, Reddi AH. Insulin-like growth factors maintain steady-state metabolism of proteoglycans in bovine articular cartilage explants. Arch Biochem Biophys. 1988 Dec;267(2):416–425. [PubMed]
  • Morales TI, Roberts AB. Transforming growth factor beta regulates the metabolism of proteoglycans in bovine cartilage organ cultures. J Biol Chem. 1988 Sep 15;263(26):12828–12831. [PubMed]
  • Sandy JD, Neame PJ, Boynton RE, Flannery CR. Catabolism of aggrecan in cartilage explants. Identification of a major cleavage site within the interglobular domain. J Biol Chem. 1991 May 15;266(14):8683–8685. [PubMed]
  • Malfait AM, Verbruggen G, Veys EM, Lambert J, De Ridder L, Cornelissen M. Comparative and combined effects of interleukin 6, interleukin 1 beta, and tumor necrosis factor alpha on proteoglycan metabolism of human articular chondrocytes cultured in agarose. J Rheumatol. 1994 Feb;21(2):314–320. [PubMed]
  • Hui W, Bell M, Carroll G. Oncostatin M (OSM) stimulates resorption and inhibits synthesis of proteoglycan in porcine articular cartilage explants. Cytokine. 1996 Jun;8(6):495–500. [PubMed]
  • Homandberg GA, Hui F, Wen C. Fibronectin fragment mediated cartilage chondrolysis. I. Suppression by anti-oxidants. Biochim Biophys Acta. 1996 Nov 15;1317(2):134–142. [PubMed]
  • Plaas AH, Sandy JD. A cartilage explant system for studies on aggrecan structure, biosynthesis and catabolism in discrete zones of the mammalian growth plate. Matrix. 1993 Mar;13(2):135–147. [PubMed]
  • Sandy JD, Flannery CR, Neame PJ, Lohmander LS. The structure of aggrecan fragments in human synovial fluid. Evidence for the involvement in osteoarthritis of a novel proteinase which cleaves the Glu 373-Ala 374 bond of the interglobular domain. J Clin Invest. 1992 May;89(5):1512–1516. [PMC free article] [PubMed]
  • Lohmander LS, Neame PJ, Sandy JD. The structure of aggrecan fragments in human synovial fluid. Evidence that aggrecanase mediates cartilage degradation in inflammatory joint disease, joint injury, and osteoarthritis. Arthritis Rheum. 1993 Sep;36(9):1214–1222. [PubMed]
  • Lark MW, Gordy JT, Weidner JR, Ayala J, Kimura JH, Williams HR, Mumford RA, Flannery CR, Carlson SS, Iwata M, et al. Cell-mediated catabolism of aggrecan. Evidence that cleavage at the "aggrecanase" site (Glu373-Ala374) is a primary event in proteolysis of the interglobular domain. J Biol Chem. 1995 Feb 10;270(6):2550–2556. [PubMed]
  • Okada Y, Shinmei M, Tanaka O, Naka K, Kimura A, Nakanishi I, Bayliss MT, Iwata K, Nagase H. Localization of matrix metalloproteinase 3 (stromelysin) in osteoarthritic cartilage and synovium. Lab Invest. 1992 Jun;66(6):680–690. [PubMed]
  • Flannery CR, Lark MW, Sandy JD. Identification of a stromelysin cleavage site within the interglobular domain of human aggrecan. Evidence for proteolysis at this site in vivo in human articular cartilage. J Biol Chem. 1992 Jan 15;267(2):1008–1014. [PubMed]
  • Bonassar LJ, Sandy JD, Lark MW, Plaas AH, Frank EH, Grodzinsky AJ. Inhibition of cartilage degradation and changes in physical properties induced by IL-1beta and retinoic acid using matrix metalloproteinase inhibitors. Arch Biochem Biophys. 1997 Aug 15;344(2):404–412. [PubMed]
  • Mort JS, Dodge GR, Roughley PJ, Liu J, Finch SJ, DiPasquale G, Poole AR. Direct evidence for active metalloproteinases mediating matrix degradation in interleukin 1-stimulated human articular cartilage. Matrix. 1993 Mar;13(2):95–102. [PubMed]
  • Buttle DJ, Handley CJ, Ilic MZ, Saklatvala J, Murata M, Barrett AJ. Inhibition of cartilage proteoglycan release by a specific inactivator of cathepsin B and an inhibitor of matrix metalloproteinases. Evidence for two converging pathways of chondrocyte-mediated proteoglycan degradation. Arthritis Rheum. 1993 Dec;36(12):1709–1717. [PubMed]
  • Loulakis P, Shrikhande A, Davis G, Maniglia CA. N-terminal sequence of proteoglycan fragments isolated from medium of interleukin-1-treated articular-cartilage cultures. Putative site(s) of enzymic cleavage. Biochem J. 1992 Jun 1;284(Pt 2):589–593. [PMC free article] [PubMed]
  • Ilic MZ, Handley CJ, Robinson HC, Mok MT. Mechanism of catabolism of aggrecan by articular cartilage. Arch Biochem Biophys. 1992 Apr;294(1):115–122. [PubMed]
  • Sandy JD, Plaas AH, Koob TJ. Pathways of aggrecan processing in joint tissues. Implications for disease mechanism and monitoring. Acta Orthop Scand Suppl. 1995 Oct;266:26–32. [PubMed]
  • Kim YJ, Sah RL, Doong JY, Grodzinsky AJ. Fluorometric assay of DNA in cartilage explants using Hoechst 33258. Anal Biochem. 1988 Oct;174(1):168–176. [PubMed]
  • Rayan V, Hardingham T. The recovery of articular cartilage in explant culture from interleukin-1 alpha: effects on proteoglycan synthesis and degradation. Matrix Biol. 1994 Apr;14(3):263–271. [PubMed]
  • Ozturk SS, Palsson BO. Chemical decomposition of glutamine in cell culture media: effect of media type, pH, and serum concentration. Biotechnol Prog. 1990 Mar-Apr;6(2):121–128. [PubMed]
  • Spencer CA, Palmer TN, Mason RM. Intermediary metabolism in the Swarm rat chondrosarcoma chondrocyte. Biochem J. 1990 Feb 1;265(3):911–914. [PMC free article] [PubMed]
  • Plagemann PG, Wohlhueter RM, Graff J, Erbe J, Wilkie P. Broad specificity hexose transport system with differential mobility of loaded and empty carrier, but directional symmetry, is common property of mammalian cell lines. J Biol Chem. 1981 Mar 25;256(6):2835–2842. [PubMed]
  • Yung S, Coles GA, Davies M. IL-1 beta, a major stimulator of hyaluronan synthesis in vitro of human peritoneal mesothelial cells: relevance to peritonitis in CAPD. Kidney Int. 1996 Oct;50(4):1337–1343. [PubMed]
  • Yagita M, Nakajima M, Saksela E. Suppression of human natural killer cell activity by amino sugars. Cell Immunol. 1989 Aug;122(1):83–95. [PubMed]
  • Tesoriere G, Vento R, Calvaruso G. Inhibitory effect of D-glucosamine on glycolysis in bovine retina. Biochim Biophys Acta. 1975 Mar 14;385(1):58–67. [PubMed]
  • Marshall S, Bacote V, Traxinger RR. Discovery of a metabolic pathway mediating glucose-induced desensitization of the glucose transport system. Role of hexosamine biosynthesis in the induction of insulin resistance. J Biol Chem. 1991 Mar 15;266(8):4706–4712. [PubMed]
  • Hayashi K, Ochiai T, Ishinoda Y, Okamoto T, Maruyama T, Tsuda K, Tsubouchi H. Relationship between cellular ATP content and cellular functions of primary cultured rat hepatocytes in hypoxia. J Gastroenterol Hepatol. 1997 Mar;12(3):249–256. [PubMed]
  • Pan YT, Kamitani T, Bhuvaneswaran C, Hallaq Y, Warren CD, Yeh ET, Elbein AD. Inhibition of glycosylphosphatidylinositol anchor formation by mannosamine. J Biol Chem. 1992 Oct 15;267(29):21250–21255. [PubMed]

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