• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. Sep 15, 2001; 358(Pt 3): 615–626.
PMCID: PMC1222096

Analysis of aggrecan in human knee cartilage and synovial fluid indicates that aggrecanase (ADAMTS) activity is responsible for the catabolic turnover and loss of whole aggrecan whereas other protease activity is required for C-terminal processing in vivo.


Studies of aggrecan proteolysis in human joints have implicated both the aggrecanase [ADAMTS, a disintegrin-like and metalloprotease (reprolysin-type) with thrombospondin type 1 motif] and matrix metalloproteinase (MMP) families. We have analysed the aggrecan core protein species present in vivo in both articular cartilage and synovial fluids from normal, acutely injured and osteoarthritic joints. Normal cartilage contains at least seven major G1 domain (the N-terminal globular domain of aggrecan)-bearing species, of which three (full-length core, G1-NITEGE(373) and G1-VDIPEN(341)) have been identified. The C-terminals of the others are unknown but digestion of fetal human aggrecan with MMP-3 and crude aggrecanase suggests that they are products of MMP-like activity in vivo. Normal synovial fluids contain at least 10 species, of which nine result from ADAMTS-dependent cleavage, and this cleavage occurs at all of the five known aggrecanase sites. Aggrecan fragments in the cartilage and synovial fluids of acutely injured joints are generally similar to normal, but all contain a markedly increased ratio of G1-NITEGE to G1-VDIPEN. Aggrecan from the cartilage of late-stage osteoarthritis patients is remarkably similar to normal, whereas the synovial fluid aggrecan is more fragmented than that from normal or injured knees. The analyses suggest that the role of the ADAMTS and these MMP-like activities in human cartilage are distinctly different. Excessive ADAMTS activity in vivo is destructive to cartilage matrix, since the bulk of the glycosaminoglycan (GAG)-bearing products are released from the tissue into the synovial fluid following cleavage of the Glu(373)-Ala(374) bond. In contrast, the MMP-like activity appears to be essentially non-destructive, since much of the GAG-bearing product is retained in the tissue following cleavages that are in the more C-terminal regions of the molecule.

Full Text

The Full Text of this article is available as a PDF (384K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Bayliss MT, Ali SY. Age-related changes in the composition and structure of human articular-cartilage proteoglycans. Biochem J. 1978 Dec 15;176(3):683–693. [PMC free article] [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]
  • Ilic MZ, Robinson HC, Handley CJ. Characterization of aggrecan retained and lost from the extracellular matrix of articular cartilage. Involvement of carboxyl-terminal processing in the catabolism of aggrecan. J Biol Chem. 1998 Jul 10;273(28):17451–17458. [PubMed]
  • Maroudas A, Bayliss MT, Uchitel-Kaushansky N, Schneiderman R, Gilav E. Aggrecan turnover in human articular cartilage: use of aspartic acid racemization as a marker of molecular age. Arch Biochem Biophys. 1998 Feb 1;350(1):61–71. [PubMed]
  • Buckwalter JA, Roughley PJ, Rosenberg LC. Age-related changes in cartilage proteoglycans: quantitative electron microscopic studies. Microsc Res Tech. 1994 Aug 1;28(5):398–408. [PubMed]
  • Roughley PJ, White RJ. The use of caesium sulphate density gradient centrifugation to analyse proteoglycans from human articular cartilages of different ages. Biochim Biophys Acta. 1983 Aug 23;759(1-2):58–66. [PubMed]
  • Dudhia J, Davidson CM, Wells TM, Vynios DH, Hardingham TE, Bayliss MT. Age-related changes in the content of the C-terminal region of aggrecan in human articular cartilage. Biochem J. 1996 Feb 1;313(Pt 3):933–940. [PMC free article] [PubMed]
  • Webber C, Glant TT, Roughley PJ, Poole AR. The identification and characterization of two populations of aggregating proteoglycans of high buoyant density isolated from post-natal human articular cartilages of different ages. Biochem J. 1987 Dec 15;248(3):735–740. [PMC free article] [PubMed]
  • Glant TT, Mikecz K, Roughley PJ, Buzás E, Poole AR. Age-related changes in protein-related epitopes of human articular-cartilage proteoglycans. Biochem J. 1986 May 15;236(1):71–75. [PMC free article] [PubMed]
  • Vilím V, Fosang AJ. Characterization of proteoglycans isolated from associative extracts of human articular cartilage. Biochem J. 1993 Jul 1;293(Pt 1):165–172. [PMC free article] [PubMed]
  • Vilim V, Fosang AJ. Proteoglycans isolated from dissociative extracts of differently aged human articular cartilage: characterization of naturally occurring hyaluronan-binding fragments of aggrecan. Biochem J. 1994 Dec 15;304(Pt 3):887–894. [PMC free article] [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]
  • 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]
  • Mort JS, Buttle DJ. The use of cleavage site specific antibodies to delineate protein processing and breakdown pathways. Mol Pathol. 1999 Feb;52(1):11–18. [PMC free article] [PubMed]
  • Little CB, Flannery CR, Hughes CE, Mort JS, Roughley PJ, Dent C, Caterson B. Aggrecanase versus matrix metalloproteinases in the catabolism of the interglobular domain of aggrecan in vitro. Biochem J. 1999 Nov 15;344(Pt 1):61–68. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Fosang AJ, Hardingham TE. Isolation of the N-terminal globular protein domains from cartilage proteoglycans. Identification of G2 domain and its lack of interaction with hyaluronate and link protein. Biochem J. 1989 Aug 1;261(3):801–809. [PMC free article] [PubMed]
  • Fosang AJ, Last K, Knäuper V, Neame PJ, Murphy G, Hardingham TE, Tschesche H, Hamilton JA. Fibroblast and neutrophil collagenases cleave at two sites in the cartilage aggrecan interglobular domain. Biochem J. 1993 Oct 1;295(Pt 1):273–276. [PMC free article] [PubMed]
  • Cole AA, Kuettner KE. MMP-8 (neutrophil collagenase) mRNA and aggrecanase cleavage products are present in normal and osteoarthritic human articular cartilage. Acta Orthop Scand Suppl. 1995 Oct;266:98–102. [PubMed]
  • Williams JM, Downey C, Thonar EJ. Increase in levels of serum keratan sulfate following cartilage proteoglycan degradation in the rabbit knee joint. Arthritis Rheum. 1988 Apr;31(4):557–560. [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]
  • Sandy JD, Westling J, Kenagy RD, Iruela-Arispe ML, Verscharen C, Rodriguez-Mazaneque JC, Zimmermann DR, Lemire JM, Fischer JW, Wight TN, et al. Versican V1 proteolysis in human aorta in vivo occurs at the Glu441-Ala442 bond, a site that is cleaved by recombinant ADAMTS-1 and ADAMTS-4. J Biol Chem. 2001 Apr 20;276(16):13372–13378. [PubMed]
  • Abbaszade I, Liu RQ, Yang F, Rosenfeld SA, Ross OH, Link JR, Ellis DM, Tortorella MD, Pratta MA, Hollis JM, et al. Cloning and characterization of ADAMTS11, an aggrecanase from the ADAMTS family. J Biol Chem. 1999 Aug 13;274(33):23443–23450. [PubMed]
  • Tortorella MD, Pratta M, Liu RQ, Austin J, Ross OH, Abbaszade I, Burn T, Arner E. Sites of aggrecan cleavage by recombinant human aggrecanase-1 (ADAMTS-4). J Biol Chem. 2000 Jun 16;275(24):18566–18573. [PubMed]
  • Kuno K, Okada Y, Kawashima H, Nakamura H, Miyasaka M, Ohno H, Matsushima K. ADAMTS-1 cleaves a cartilage proteoglycan, aggrecan. FEBS Lett. 2000 Aug 4;478(3):241–245. [PubMed]
  • Bayliss MT, Roughley PJ. The properties of proteoglycan prepared from human articular cartilage by using associative caesium chloride gradients of high and low starting densities. Biochem J. 1985 Nov 15;232(1):111–117. [PMC free article] [PubMed]
  • Fosang AJ, Last K, Maciewicz RA. Aggrecan is degraded by matrix metalloproteinases in human arthritis. Evidence that matrix metalloproteinase and aggrecanase activities can be independent. J Clin Invest. 1996 Nov 15;98(10):2292–2299. [PMC free article] [PubMed]
  • Lee ER, Lamplugh L, Leblond CP, Mordier S, Magny MC, Mort JS. Immunolocalization of the cleavage of the aggrecan core protein at the Asn341-Phe342 bond, as an indicator of the location of the metalloproteinases active in the lysis of the rat growth plate. Anat Rec. 1998 Sep;252(1):117–132. [PubMed]
  • Mort JS, Magny MC, Lee ER. Cathepsin B: an alternative protease for the generation of an aggrecan 'metalloproteinase' cleavage neoepitope. Biochem J. 1998 Nov 1;335(Pt 3):491–494. [PMC free article] [PubMed]
  • Sandy JD, Thompson V, Doege K, Verscharen C. The intermediates of aggrecanase-dependent cleavage of aggrecan in rat chondrosarcoma cells treated with interleukin-1. Biochem J. 2000 Oct 1;351(Pt 1):161–166. [PMC free article] [PubMed]
  • Sandy JD, Plaas AH. Studies on the hyaluronate binding properties of newly synthesized proteoglycans purified from articular chondrocyte cultures. Arch Biochem Biophys. 1989 Jun;271(2):300–314. [PubMed]
  • Sandy JD, Gamett D, Thompson V, Verscharen C. Chondrocyte-mediated catabolism of aggrecan: aggrecanase-dependent cleavage induced by interleukin-1 or retinoic acid can be inhibited by glucosamine. Biochem J. 1998 Oct 1;335(Pt 1):59–66. [PMC free article] [PubMed]
  • Farndale RW, Buttle DJ, Barrett AJ. Improved quantitation and discrimination of sulphated glycosaminoglycans by use of dimethylmethylene blue. Biochim Biophys Acta. 1986 Sep 4;883(2):173–177. [PubMed]
  • Bonassar LJ, Frank EH, Murray JC, Paguio CG, Moore VL, Lark MW, Sandy JD, Wu JJ, Eyre DR, Grodzinsky AJ. Changes in cartilage composition and physical properties due to stromelysin degradation. Arthritis Rheum. 1995 Feb;38(2):173–183. [PubMed]
  • Fosang AJ, Last K, Knäuper V, Murphy G, Neame PJ. Degradation of cartilage aggrecan by collagenase-3 (MMP-13). FEBS Lett. 1996 Feb 12;380(1-2):17–20. [PubMed]
  • Lark MW, Bayne EK, Flanagan J, Harper CF, Hoerrner LA, Hutchinson NI, Singer II, Donatelli SA, Weidner JR, Williams HR, et al. Aggrecan degradation in human cartilage. Evidence for both matrix metalloproteinase and aggrecanase activity in normal, osteoarthritic, and rheumatoid joints. J Clin Invest. 1997 Jul 1;100(1):93–106. [PMC free article] [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]
  • Dahlberg L, Fridén T, Roos H, Lark MW, Lohmander LS. A longitudinal study of cartilage matrix metabolism in patients with cruciate ligament rupture--synovial fluid concentrations of aggrecan fragments, stromelysin-1 and tissue inhibitor of metalloproteinase-1. Br J Rheumatol. 1994 Dec;33(12):1107–1111. [PubMed]
  • Sztrolovics R, Alini M, Roughley PJ, Mort JS. Aggrecan degradation in human intervertebral disc and articular cartilage. Biochem J. 1997 Aug 15;326(Pt 1):235–241. [PMC free article] [PubMed]
  • Plaas AH, West LA, Wong-Palms S, Nelson FR. Glycosaminoglycan sulfation in human osteoarthritis. Disease-related alterations at the non-reducing termini of chondroitin and dermatan sulfate. J Biol Chem. 1998 May 15;273(20):12642–12649. [PubMed]
  • Brown MP, West LA, Merritt KA, Plaas AH. Changes in sulfation patterns of chondroitin sulfate in equine articular cartilage and synovial fluid in response to aging and osteoarthritis. Am J Vet Res. 1998 Jun;59(6):786–791. [PubMed]
  • Doege KJ, Sasaki M, Kimura T, Yamada Y. Complete coding sequence and deduced primary structure of the human cartilage large aggregating proteoglycan, aggrecan. Human-specific repeats, and additional alternatively spliced forms. J Biol Chem. 1991 Jan 15;266(2):894–902. [PubMed]

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


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...