Degradation of small leucine-rich repeat proteoglycans by matrix metalloprotease-13: identification of a new biglycan cleavage site
© Monfort et al.; licensee BioMed Central Ltd. 2006
Received: 4 August 2005
Accepted: 28 November 2005
Published: 3 January 2006
The Erratum to this article has been published in Arthritis Research & Therapy 2013 15:401
A major and early feature of cartilage degeneration is proteoglycan breakdown. Matrix metalloprotease (MMP)-13 plays an important role in cartilage degradation in osteoarthritis (OA). This MMP, in addition to initiating collagen fibre cleavage, acts on several proteoglycans. One of the proteoglycan families, termed small leucine-rich proteoglycans (SLRPs), was found to be involved in collagen fibril formation/interaction, with some members playing a role in the OA process. We investigated the ability of MMP-13 to cleave members of two classes of SLRPs: biglycan and decorin; and fibromodulin and lumican. SLRPs were isolated from human normal and OA cartilage using guanidinium chloride (4 mol/l) extraction. Digestion products were examined using Western blotting. The identities of the MMP-13 degradation products of biglycan and decorin (using specific substrates) were determined following electrophoresis and microsequencing. We found that the SLRPs studied were cleaved to differing extents by human MMP-13. Although only minimal cleavage of decorin and lumican was observed, cleavage of fibromodulin and biglycan was extensive, suggesting that both molecules are preferential substrates. In contrast to biglycan, decorin and lumican, which yielded a degradation pattern similar for both normal and OA cartilage, fibromodulin had a higher level of degradation with increased cartilage damage. Microsequencing revealed a novel major cleavage site (... G177/V178) for biglycan and a potential cleavage site for decorin upon exposure to MMP-13. We showed, for the first time, that MMP-13 can degrade members from two classes of the SLRP family, and identified the site at which biglycan is cleaved by MMP-13. MMP-13 induced SLRP degradation may represent an early critical event, which may in turn affect the collagen network by exposing the MMP-13 cleavage site in this macromolecule. Awareness of SLRP degradation products, especially those of biglycan and fibromodulin, may assist in early detection of OA cartilage degradation.
Osteoarthritis (OA) is the most common rheumatologic disease, with high incidence and morbidity. Even though the early pathophysiological process remains to be elucidated, one of the first alterations in OA cartilage is a decrease in proteoglycan content . Proteoglycans form a large group that can be classified into five families according to the structural properties of their core protein . One group, termed the small leucine-rich proteoglycans (SLRPs), possesses a central domain of characteristic repeats that participate in protein-protein interactions . The SLRPs can be divided into four classes based on gene organization and amino acid sequence homologies : class I includes decorin, biglycan and asporin; class II includes fibromodulin, lumican, keratocan, PRELP (proline arginine-rich end leucine-rich repeat protein) and osteoadherin; class III includes epiphycan, mimecan and opticin; and class IV includes chondroadherin and the recently identified nyctalopin .
Although an understanding of the functions of SLRPs is only now emerging, most of the members bind specifically to other extracellular matrix constituents and contribute to the structural framework of connective tissues . Moreover, some were shown to interact with various collagen types, including collagen type II, and to influence collagen fibril formation and interaction. These include decorin , fibromodulin , asporin , lumican , PRELP  and chondroadherin . Moreover, fibromodulin, asporin, biglycan, decorin and lumican were also suggested to play a role in the OA cartilage process [11–13].
Decorin was the first in this series of molecules to be structurally defined. It contains one glycosaminoglycan chain, often dermatan sulfate, which can adopt complex secondary structures and form specific interactions with matrix molecules . The decorin level in cartilage is by far the most abundant of the SLRPs, and in humans its level increases with increasing age . Its proposed major functions are the regulation of collagen fibrillogenesis and maintenance of tissue integrity by its binding with fibronectin and thrombospondin [15–17] The closely related family member biglycan, despite its 57% of homology with decorin , does not interact with collagen under all conditions. Biglycan interactions appear to be primarily with type VI collagen. Biglycan has been identified at the surface of cartilage and in the pericellular region. In OA cartilage, a higher concentration was reported in the deeper layers of the tissue .
Fibromodulin contains up to four keratan sulphate chains  and was originally described as a collagen-binding protein. It is able to influence collagen fibril formation and maintain a sustained interaction with the formed fibrils . Lumican, which is present at a high level in the cornea , has a widespread distribution in connective tissues [5, 22, 23], including cartilage . Lumican and fibromodulin have been shown to bind to the same site on the collagen fibril [20, 25]. Lumican modulates collagen fibrillogenesis and enhances collagen fibril stability .
Synthesis of collagen in normal and pathological cartilage is slow. However, in OA the integrity of the collagen network is impaired. This could result from defective linking of the collagen fibrils by molecules such as the SLRPs, thus interfering with the network stability, preventing its repair and accelerating its degradation. Cleavage of the SLRPs may then precede major destruction of the collagen and contribute to this process . Data in the literature show that members of the matrix metalloprotease (MMP) family are able to cleave some SLRPs. MT1-MMP can cleave human recombinant lumican ; MMP-2, MMP-3 and MMP-7 cleave human recombinant decorin ; and MMP-13 cleaves bovine fibromodulin when this molecule is bound to collagen . Purified bovine fibromodulin cannot be cleaved by human MMP-13 . It was also recently shown that truncated disintegrin-like and metalloprotease domain with thrombospondin type I motifs-4 (ADAMTS-4) can cleave the MMP-13 susceptible bond of fibromodulin . However, MMP-2, MMP-8 and MMP-9 do not cleave fibromodulin .
Although various MMPs are present in human OA cartilage, MMP-13 was demonstrated to play a major role. This enzyme, in addition to cleaving native collagen and having a higher activity on type II collagen than MMP-1, also acts to degrade various extracellular macromolecules including proteoglycans . However, limited studies have been done on its effect on the SLRPs. We therefore investigated the ability of human recombinant MMP-13 to cleave members of two classes of the SLRPs (class I decorin and biglycan, and class II fibromodulin and lumican), derived from normal and OA human cartilage differing in the severity of the disease process. The results show that MMP-13 can degrade all four SLRPs, with fibromodulin and biglycan being preferential substrates.
Materials and methods
Normal human cartilage (femoral condyles and tibial plateaus) was obtained from individuals within 12 hours of death at time of autopsy (n = 3; mean age [± standard deviation] 52 ± 14 years). These individuals had no history of joint disease and died from causes unrelated to arthritic diseases, including cardiorespiratory arrest, cerebral haemorrhage and pulmonary embolism. The tissue was examined macroscopically and histologically to ensure that only normal tissue was used.
OA human cartilage (femoral condyles and tibial plateaus) was obtained from patients undergoing total knee arthroplasty (n = 9; mean age [± standard deviation] 76 ± 5 years). All patients were evaluated by a certified rheumatologist who used the American College of Rheumatology criteria for OA of the knee . These specimens represented early, moderate, or severe OA, as defined by microscopic criteria [31–33]. The Clinical Research Ethics Committee of the University of Montreal Hospital Center approved the study protocol and the use of human tissues.
Proteoglycans were extracted with 4 mol/l guanidinium chloride [34, 35]. Briefly, cartilage was finely diced to pieces and extracted with 4 mol/l guanidinium chloride (Invitrogen Inc., Carlsbad, CA, USA) in 0.1 mol/l sodium acetate (pH 6.0) containing protease inhibitors (leupeptin [10 μg/ml], pepstatin [10 μg/ml], aprotinin [10 μg/ml], 1,10-phenanthroline [10 μg/ml] and phenylmethanesulphonyl fluoride [100 μg/ml]; EMD Biosciences Inc., La Jolla, CA, USA) at 4°C with continuous stirring for 48 hours. The extract was then separated from the cartilage residue by filtration through glass wool, and then dialyzed for 48 hours against 50 mmol/l Tris buffer (pH 7.5). One might argue that because the inhibitors were removed during the dialysis the endogenous MMPs could have been activated. However, because 1,10-phenanthroline is a zinc chelator, the catalytic zinc would also be removed by the dialysis, and so the MMPs would remain inactive.
Analysis of SLRP cleavage by MMP-13
MMP-13 proteolytic activity was analyzed on human normal (n = 3) and OA cartilage having different levels of fibrillation corresponding to the different stage of the disease process. These were named slightly (n = 3), moderately (n = 3) and severely (n = 3) fibrillated cartilage. Proteoglycan extracts were incubated for 0–16 hours with human recombinant (rh)MMP-13 (R&D Systems Inc., Minneapolis, MN, USA) activated with 0.5 mmol/l aminophenylmercuric acetate (APMA; Kodak Inc., Toronto, ON, Canada) in 50 mmol/l Tris-HCl (pH 7.5) containing 10 mmol/l CaCl2 and 0.05% Brij 35 (Sigma-Aldrich Canada Ltd., Oakville, ON, Canada) at an MMP-13/proteoglycan ratio of 1:50 (100 ng/5 μg). Glycosaminoglycan content was determined using the 1,2-dimethylmethylene blue (DMMB) method . The reaction was stopped by the addition of EDTA (Sigma-Aldrich Canada Ltd.) at a final concentration of 15 mmol/l. The samples were treated with 25 mU chondroitinase ABC (#C-2905; Sigma-Aldrich Canada Ltd.)/100 μl proteoglycan extract overnight at 37°C. In addition, a control was performed with the moderately fibrillated cartilage in which no MMP-13 was added and samples were incubated for 16 hours. Data were identical to those with the nonincubated specimens (data not shown).
In order to investigate MMP-13 specificity, RS 110–2481 (a synthetic specific MMP-13 carboxylate inhibitor generously provided by C Myers [Roche Bioscience, Palo Alto, CA, USA]) , was used. The Ki (nmol/l) for MMP-1, MMP-2, MMP-3, MMP-8 and MMP-13 were 1:100, 32, 19, 18 and 0.08, respectively. Briefly, samples from moderately fibrillated cartilage extract were treated with rhMMP-13 and RS 110–2481 at 1 and 50 nmol/l for the indicated time, and samples processed for Western blotting.
Proteoglycan solutions were mixed with a sample buffer (62.5 mmol/l Tris-HCl [pH 6.8], 2% w/v sodium dodecyl sulphate, 10% glycerol, 5% β-mercaptoethanol, and 0.05% bromophenol blue) and electrophoresed on 4–20% Ready-Gels (Bio-Rad Laboratories Ltd., Mississauga, ON, Canada). They were then transferred electrophoretically to nitrocellulose membranes (Bio-Rad Laboratories Ltd.) and processed for Western immunoblotting. Blots were blocked in 2% low fat dry milk in Tris-buffered saline containing 0.05% Tween 20 (Sigma-Aldrich Canada Ltd.). As described previously , rabbit polyclonal antibodies raised against synthetic peptides corresponding to the carboxyl-terminus of the SLRP core proteins were used as primary antibodies for the detection of biglycan (1/5,000 dilution), fibromodulin (1/10,000 dilution), lumican (1/5,000 dilution) and decorin (1/5,000 dilution). The second antibody was a horseradish peroxidase-conjugated goat anti-rabbit immunoglobulin (1/10,000 dilution; Pierce, Rockford, IL, USA). Detection was performed by chemiluminescence using the Super Signal® ULTRA chemiluminescent substrate (Pierce), in accordance with the manufacturer's specifications.
Sequencing of biglycan and decorin degradation products
Bovine recombinant biglycan (15 μg) and decorin (15 μg; Sigma-Aldrich Canada Ltd.) were incubated for 1 hour at 37°C with APMA-activated rhMMP-13 in 50 mmol/l Tris-HCl (pH 7.5), containing 10 nmol/l CaCl2 and 0.05% Brij 35. The reaction was stopped by the addition of EDTA at a final concentration of 15 mmol/l. Glycosaminoglycan chains were removed by incubation with 0.1 unit chondroitinase ABC (#C-3667; Sigma-Aldrich Canada Ltd.) for 8 hours at room temperature, followed by boiling for 5 minutes with the electrophoresis sample buffer. To remove Asn-linked oligosaccharides, N-glycanase (0.3 unit; Roche Diagnostics, Laval, QC, Canada) and sample buffer containing 1.2% Nonidet P-40 (Roche Diagnostics) were added to the solution, which was then incubated again for 12 hours at room temperature. Degradation products were separated in 4–20% polyacrylamide gels (Bio-Rad Laboratories Ltd.). After electrophoresis, the gels were soaked in CAPS transfer buffer (10 nmol/l 3-cyclohexylamino-1-propanesulfonic acid, 10% methanol; pH 11.0) for 15 minutes at 0.25 A. After washing, the proteins were transferred onto PVDF membranes (Millipore Corporation, Bedford, MA, USA), which were washed in de-ionized water, stained with 0.1% Coomassie Blue in 50% methanol for 5 minutes, and then de-stained in 50% methanol and 10% acetic acid for 5–7 minutes at room temperature. Finally, the membrane was rinsed in de-ionized water, air dried and stored at room temperature. Amino-terminal amino acid sequencing of the protein band was performed on a Procise Protein Sequencer model 492 (Applied Biosystems, Foster City, CA, USA).
The use of human cartilage extracts to analyze SLRP degradation allowed study of all four SLRPs in a single extract under identical conditions, and permitted SLRP degradation to be carried out in a physiologically relevant extract of matrix proteins.
MMP-13 degrades biglycan and decorin
To determine whether MMP-13 was the sole enzyme responsible for the cleavage, and not other enzymes present in the cartilage extracts, we further treated the samples from the moderately fibrillated cartilage with two concentrations (1 and 50 nmol/l) of a preferential inhibitor of MMP-13, namely RS 110–2481 . Biglycan degradation was completely prevented at both concentrations tested (Figure 1e).
MMP-13 cleavage sites of biglycan and decorin
Amino acid sequencing analysis was performed with recombinant biglycan and decorin treated with MMP-13. In contrast to the Western blotting, which identifies carboxyl-terminal fragments, sequence analysis can identify the amino-terminus of all fragments.
Sequence analysis of the two decorin cleavage fragments of 28 and 26 kDa showed that they possessed the same amino-terminus. The larger fragment is compatible with cleavage between positions 240 and 241 of the peptidic chain corresponding to a previously reported  cleavage site between the serine (S) and leucine (L). The exact cleavage site of the smaller fragment could not be identified.
The SLRP fragment sizes visualized on the gel used for sequencing were smaller than those observed on the gel used for Western blotting, possibly due to the treatment with N-glycanase in the former procedure. Of note, molecular weight determination by Western blotting is an approximation.
Degradation of fibromodulin and lumican
A major and early feature of cartilage degeneration is proteoglycan breakdown. MMP-13 has been shown to play an important role in OA cartilage degeneration by its effect not only on the collagen network but also on proteoglycans . In the present study we investigated the ability of human MMP-13 to act on members of the SLRP proteoglycan family derived from human cartilage ranging from normal to advanced OA.
One emerging observation is that biglycan and fibromodulin are preferential substrates for MMP-13, whereas degradation of decorin and lumican is much less effective. This could imply that biglycan and fibromodulin are sensitive to both the gelatinolytic and collagenolytic activities of MMP-13, whereas decorin and lumican are more responsive to the gelatinolytic cleavage. Support for this hypothesis was provided by Imai and colleagues , who showed that decorin could be cleaved by MMP-2, MMP-3 and MMP-7, whereas cleavage with MMP-1 was negligible. The greater effect of MMP-13 than of MMP-1 on decorin could be due to the fact that the former enzyme has 44 times more gelatinolytic activity than does MMP-1 . Moreover, and in agreement with this hypothesis, only 1 nmol/l of the inhibitor RS 110–2481 is sufficient to prevent collagenolytic activity, but 50 nmol/l is required to prevent gelatinolytic activity , and the effect of MMP-13 on biglycan and fibromodulin is abolished at both inhibitor concentrations whereas the effect on decorin and lumican is abolished only at the higher concentration.
Biglycan is found in the pericellular matrix of many connective tissues, and appears to play a role in regulating morphogenesis and differentiation . Although biglycan is present in cartilage and is upregulated in the late stages of OA , its exact role in OA remains to be determined. The present data show that in some specimens a biglycan fragment of a similar size to that generated by MMP-13 is present in the cartilage as a minor component. It is possible that this in situ degradation product might not be cleaved at exactly the same site. This requires further study with an antibody recognizing the amino-terminal sequence of the fragment; however, such an antibody is not yet available. It is also possible that the biglycan degradation product may not be stably retained within the cartilage matrix and hence may not accumulate in large amounts. The study showed that the degree of biglycan degradation was independent of the extent of cartilage damage, although the amount of biglycan present in the severely fibrillated cartilage was significantly less than in normal to moderately fibrillated specimens. This suggests that, in the severely fibrillated specimens, biglycan has already been extensively degraded, leading to the loss of the epitope recognized by the antibody. Although we cannot exclude the possibility that proteases other than MMP-13 exerted an affect on this SLRP, this is unlikely because all endogenous carboxy, serine and MMPs should have been irreversibly inhibited by the inhibitor cocktail used in the extraction procedure. Although some cysteine proteases may survive the extraction procedure, it is unlikely that they remain active at pH 7.5, which was used for the incubation.
Our data also showed that MMP-13 induces two main biglycan fragments. The larger fragment possessed a new cleavage site (... G177-V178) in the leucine-rich region. The second smaller fragment possessed the same carboxyl-terminal sequence, indicating the presence of a second cleavage site. As the antibody used for immunodetection recognizes the carboxyl-terminal region of biglycan, cleavage at this second site must be after the G177-V178 cleavage site found in the larger fragment.
As mentioned above, Imai and colleagues  demonstrated the ability of three MMPs – namely MMP-2, MMP-3 and MMP-7 – to degrade decorin, and reported multiple cleavage sites. It seems likely that these MMPs cleaved within the leucine-rich region at different sites, because all fragments, albeit of different sizes, possessed the same amino-terminal sequence corresponding to that of the intact decorin core protein . The present study revealed that MMP-13 degrades decorin into two fragments that also possess the same amino-terminal sequence as the intact decorin core protein. The products identified by amino acid sequencing from recombinant decorin were of 28 and 26 kDa. These may represent the amino-terminal fragments corresponding to the cartilage extract decorin fragments identified with a carboxyl-terminal antibody, because it appears that decorin cleavage occurs toward the centre of the molecule. One would expect the amino-terminal and carboxyl-terminal fragments to be of similar size. Because the degradation of decorin by MMP-13 appears to be due to its gelatinase activity rather than its collagenase activity, it is likely that one of the MMP-13 cleavages could be at the S240-L241 site, which is the cleavage used by gelatinase A (MMP-2) , and the other fragment would then be due to a cleavage amino-terminal of this site. This S240-L241 cleavage site is very plausible for MMP-13, because it is between aliphatic and hydrophobic amino acids, which are preferred by MMPs .
Interestingly, one of the characteristics of decorin is its interaction with active transforming growth factor (TGF)-β, thereby providing a tissue reservoir of this factor . Our data showing MMP-13 cleavage in the leucine-rich repeats suggests the possibility that TGF-β may be released from the decorin after digestion with this MMP. We recently reported that, in OA cartilage, the TGF-β level is upregulated and responsible for the in situ increase in MMP-13 in this disease tissue [42, 43]. The effect of MMP-13 on decorin, although not a preferential substrate, could be threefold. It may permit collagen degradation by its loss from the surface of the collagen fibrils; since data suggest that the leucine-rich repeats play a critical role in the interaction of SLRPs with collagens , it may result in loss of tissue integrity through the functional failure of decorin and biglycan interactions; and it may promote tissue degradation via TGF-β release, leading to increased MMP-13 production.
Lumican was reported to be present in human cartilage , but no direct evidence of its involvement in human OA has yet been reported. However, Young and colleagues  recently showed that lumican is upregulated in an ovine meniscectomy model of OA. This upregulated expression in degenerative cartilage was associated with increased lumican core protein deficient in keratan sulphate chains . The present study showed that lumican degradation by MMP-13 occurs after an incubation period of 16 hours. This appeared independent of the level of fibrillation of the cartilage from which it was extracted, indicating that lumican degradation is independent of interactions with the various components in the different cartilage extracts.
Fibromodulin cleavage by MMP-13 has previously been demonstrated . In human fibromodulin, cleavage occurs at the Y63-T64 site in the amino-terminal region of the molecule. In the present study MMP-13 degradation of fibromodulin generated a fragment of 30 kDa, which presumably corresponds to the fragment described by Heathfield and colleagues . Of note, this fragment is generated in moderately and severely fibrillated cartilage, but not in normal or slightly fibrillated cartilage, reflecting an increased sensitivity of fibromodulin to degradation when the cartilage is more degenerated. This could be related to the presence of other components in the cartilage extracts that interact with the fibromodulin. Varying abundance of such components between the differently affected cartilages could then influence MMP-13 cleavage. The work by Heathfield and colleagues  suggests that cleavage of fibromodulin is dependent on its ability to bind type II collagen. There are two possibilities that could explain this situation. First, the ability of isolated SLRPs to interact with one another could result in the cleavage site being hidden. The recent description of decorin adopting a dimeric conformation in both the solution and crystal state may relate to this hypothesis, if other SLRPs behave in a similar manner . It is possible that this dimeric conformation is removed when the SLRP binds to collagen and the MMP-13 cleavage site is then exposed. A second hypothesis could be that isolated SLRPs can act as zinc-binding proteins . If this is a property of only free SLRPs, then in the absence of collagen or other binding partner the molecules could remove the zinc site necessary for MMP-13 function.
Although MMP-13 was shown to degrade type II collagen fibrils efficiently , it is possible that in vivo SLRP interaction may help to protect the fibrils by impeding access to the collagenase cleavage site. Data from this study are of importance in human OA pathophysiology, because MMP-13-induced SLRP degradation may represent an initial event in collagen fibril degradation, by exposing the collagen fibrils to proteolytic attack and permitting subsequent cartilage degeneration. In vivo identification of the SLRP degradation products, especially those of biglycan and fibromodulin, may assist in early detection of degeneration in OA cartilage.
In this study we demonstrated the ability of human recombinant MMP-13 to cleave members of two classes of SLRPs (decorin, biglycan, fibromodulin and lumican) derived from normal and OA human cartilage differing in severity of the disease process. Although minimal cleavage of decorin and lumican was observed, cleavage of fibromodulin and biglycan was extensive, suggesting that both molecules are preferential substrates. We demonstrated that fibromodulin has a higher level of degradation with increased cartilage damage. We also characterized a novel major cleavage site for biglycan. We hypothesized that MMP-13-induced SLRP degradation may represent an early critical event in the process of cartilage degradation. Awareness of the SLRP degradation products may assist in early detection of OA cartilage degradation.
= aminophenylmercuric acetate
= matrix metalloprotease
= proline arginine-rich end leucine-rich repeat protein
= human recombinant
= small leucine-rich proteoglycan
= transforming growth factor.
We would like to thank Christelle Boileau, PhD, Alexander Watson, BSc, Changshan Geng, MD, MSc, David Hum, MSc, and François Jolicoeur, MSc, for their outstanding technical support; Pierre Pépin, MSc, from Sheldon Biotechnology for his assistance in protein sequencing; and C Myers from Roche Bioscience, Palo Alto, CA, USA for providing the MMP-13 inhibitor. The authors also thank Santa Fiori and Virginia Wallis for their assistance in manuscript preparation.
- Heinegard D, Bayliss M, Lorenzo P: Pathogenesis of structural changes in the osteoarthritic joint. Osteoarthritis. Edited by: Brandt KD, Doherty M, Lohmander SL. 2003, New York: Oxford University Press Inc, 73-92.Google Scholar
- Kjellen L, Lindahl U: Proteoglycans: structures and interactions. Annu Rev Biochem. 1991, 60: 443-475. 10.1146/annurev.bi.60.070191.002303.View ArticlePubMedGoogle Scholar
- Poole AR: Cartilage in Health and Disease. Arthritis and Allied Conditions. Edited by: Koopman WJ, Moreland LW. 2005, Philadelphia: Lippincott, Williams & Wilkins, 223-269.Google Scholar
- Bech-Hansen NT, Naylor MJ, Maybaum TA, Sparkes RL, Koop B, Birch DG, Bergen AA, Prinsen CF, Polomeno RC, Gal A, et al: Mutations in NYX, encoding the leucine-rich proteoglycan nyctalopin, cause X-linked complete congenital stationary night blindness. Nat Genet. 2000, 26: 319-323. 10.1038/81619.View ArticlePubMedGoogle Scholar
- Knudson CB, Knudson W: Cartilage proteoglycans. Semin Cell Dev Biol. 2001, 12: 69-78. 10.1006/scdb.2000.0243.View ArticlePubMedGoogle Scholar
- Hedlund H, Mengarelli-Widholm S, Heinegard D, Reinholt FP, Svensson O: Fibromodulin distribution and association with collagen. Matrix Biol. 1994, 14: 227-232. 10.1016/0945-053X(94)90186-4.View ArticlePubMedGoogle Scholar
- Lorenzo P, Aspberg A, Onnerfjord P, Bayliss MT, Neame PJ, Heinegard D: Identification and characterization of asporin. A novel member of the leucine-rich repeat protein family closely related to decorin and biglycan. J Biol Chem. 2001, 276: 12201-12211. 10.1074/jbc.M010932200.View ArticlePubMedGoogle Scholar
- Sztrolovics R, White RJ, Poole AR, Mort JS, Roughley PJ: Resistance of small leucine-rich repeat proteoglycans to proteolytic degradation during interleukin-1-stimulated cartilage catabolism. Biochem J. 1999, 339: 571-577. 10.1042/0264-6021:3390571.PubMed CentralView ArticlePubMedGoogle Scholar
- Bengtsson E, Morgelin M, Sasaki T, Timpl R, Heinegard D, Aspberg A: The leucine-rich repeat protein PRELP binds perlecan and collagens and may function as a basement membrane anchor. J Biol Chem. 2002, 277: 15061-15068. 10.1074/jbc.M108285200.View ArticlePubMedGoogle Scholar
- Mansson B, Wenglen C, Morgelin M, Saxne T, Heinegard D: Association of chondroadherin with collagen type II. J Biol Chem. 2001, 276: 32883-32888. 10.1074/jbc.M101680200.View ArticlePubMedGoogle Scholar
- Young AA, Smith MM, Smith SM, Cake MA, Ghosh P, Read RA, Melrose J, Sonnabend DH, Roughley PJ, Little CB: Regional assessment of articular cartilage gene expression and small proteoglycan metabolism in an animal model of osteoarthritis. Arthritis Res Ther. 2005, 7: R852-R861. 10.1186/ar1756.PubMed CentralView ArticlePubMedGoogle Scholar
- Kizawa H, Kou I, Iida A, Sudo A, Miyamoto Y, Fukuda A, Mabuchi A, Kotani A, Kawakami A, Yamamoto S, et al: An aspartic acid repeat polymorphism in asporin inhibits chondrogenesis and increases susceptibility to osteoarthritis. Nat Genet. 2005, 37: 138-144. 10.1038/ng1496.View ArticlePubMedGoogle Scholar
- Bock HC, Michaeli P, Bode C, Schultz W, Kresse H, Herken R, Miosge N: The small proteoglycans decorin and biglycan in human articular cartilage of late-stage osteoarthritis. Osteoarthritis Cartilage. 2001, 9: 654-663. 10.1053/joca.2001.0420.View ArticlePubMedGoogle Scholar
- Melching LI, Roughley PJ: The synthesis of dermatan sulphate proteoglycans by fetal and adult human articular cartilage. Biochem J. 1989, 261: 501-508.PubMed CentralView ArticlePubMedGoogle Scholar
- Imai K, Hiramatsu A, Fukushima D, Pierschbacher MD, Okada Y: Degradation of decorin by matrix metalloproteinases: identification of the cleavage sites, kinetic analyses and transforming growth factor-beta1 release. Biochem J. 1997, 322: 809-814.PubMed CentralView ArticlePubMedGoogle Scholar
- Winnemoller M, Schon P, Vischer P, Kresse H: Interactions between thrombospondin and the small proteoglycan decorin: interference with cell attachment. Eur J Cell Biol. 1992, 59: 47-55.PubMedGoogle Scholar
- Winnemoller M, Schmidt G, Kresse H: Influence of decorin on fibroblast adhesion to fibronectin. Eur J Cell Biol. 1991, 54: 10-17.PubMedGoogle Scholar
- Iozzo RV: Matrix proteoglycans: from molecular design to cellular function. Annu Rev Biochem. 1998, 67: 609-652. 10.1146/annurev.biochem.67.1.609.View ArticlePubMedGoogle Scholar
- Poole AR, Rosenberg LC, Reiner A, Ionescu M, Bogoch E, Roughley PJ: Contents and distributions of the proteoglycans decorin and biglycan in normal and osteoarthritic human articular cartilage. J Orthop Res. 1996, 14: 681-689. 10.1002/jor.1100140502.View ArticlePubMedGoogle Scholar
- Heathfield TF, Onnerfjord P, Dahlberg L, Heinegard D: Cleavage of fibromodulin in cartilage explants involves removal of the N-terminal tyrosine sulfate-rich region by proteolysis at a site that is sensitive to matrix metalloproteinase-13. J Biol Chem. 2004, 279: 6286-6295. 10.1074/jbc.M307765200.View ArticlePubMedGoogle Scholar
- Chakravarti S, Stallings RL, SundarRaj N, Cornuet PK, Hassell JR: Primary structure of human lumican (keratan sulfate proteoglycan) and localization of the gene (LUM) to chromosome 12q21.3-q22. Genomics. 1995, 27: 481-488. 10.1006/geno.1995.1080.View ArticlePubMedGoogle Scholar
- Iozzo RV, Murdoch AD: Proteoglycans of the extracellular environment: clues from the gene and protein side offer novel perspectives in molecular diversity and function. FASEB J. 1996, 10: 598-614.PubMedGoogle Scholar
- Hocking AM, Shinomura T, McQuillan DJ: Leucine-rich repeat glycoproteins of the extracellular matrix. Matrix Biol. 1998, 17: 1-19. 10.1016/S0945-053X(98)90121-4.View ArticlePubMedGoogle Scholar
- Grover J, Chen XN, Korenberg JR, Roughley PJ: The human lumican gene. Organization, chromosomal location, and expression in articular cartilage. J Biol Chem. 1995, 270: 21942-21949. 10.1074/jbc.270.37.21942.View ArticlePubMedGoogle Scholar
- Svensson L, Narlid I, Oldberg A: Fibromodulin and lumican bind to the same region on collagen type I fibrils. FEBS Lett. 2000, 470: 178-182. 10.1016/S0014-5793(00)01314-4.View ArticlePubMedGoogle Scholar
- Chakravarti S, Magnuson T, Lass JH, Jepsen KJ, LaMantia C, Carroll H: Lumican regulates collagen fibril assembly: skin fragility and corneal opacity in the absence of lumican. J Cell Biol. 1998, 141: 1277-1286. 10.1083/jcb.141.5.1277.PubMed CentralView ArticlePubMedGoogle Scholar
- Li Y, Aoki T, Mori Y, Ahmad M, Miyamori H, Takino T, Sato H: Cleavage of lumican by membrane-type matrix metalloproteinase-1 abrogates this proteoglycan-mediated suppression of tumor cell colony formation in soft agar. Cancer Res. 2004, 64: 7058-7064. 10.1158/0008-5472.CAN-04-1038.View ArticlePubMedGoogle Scholar
- Kashiwagi M, Enghild JJ, Gendron C, Hughes C, Caterson B, Itoh Y, Nagase H: Altered proteolytic activities of ADAMTS-4 expressed by C-terminal processing. J Biol Chem. 2004, 279: 10109-10119. 10.1074/jbc.M312123200.View ArticlePubMedGoogle Scholar
- Fosang AJ, Last K, Knauper V, Murphy G, Neame PJ: Degradation of cartilage aggrecan by collagenase-3 (MMP-13). FEBS Lett. 1996, 380: 17-20. 10.1016/0014-5793(95)01539-6.View ArticlePubMedGoogle Scholar
- Altman RD, Asch E, Bloch DA, Bole G, Borenstein D, Brandt KD, Christy W, Cooke TD, Greenwald R, Hochberg M, et al: Development of criteria for the classification and reporting of osteoarthritis. Classification of osteoarthritis of the knee. Arthritis Rheum. 1986, 29: 1039-1049.View ArticlePubMedGoogle Scholar
- Mankin HJ, Dorfman H, Lippiello L, Zarins A: Biochemical and metabolic abnormalities in articular cartilage from osteoarthritic human hips. II. Correlation of morphology with biochemical and metabolic data. J Bone Joint Surg Am. 1971, 53: 523-537.PubMedGoogle Scholar
- Pelletier JP, Martel-Pelletier J, Howell DS, Ghandur-Mnaymneh L, Enis JE, Woessner JF: Collagenase and collagenolytic activity in human osteoarthritic cartilage. Arthritis Rheum. 1983, 26: 63-68.View ArticlePubMedGoogle Scholar
- Martel-Pelletier J, Pelletier JP, Cloutier JM, Howell DS, Ghandur-Mnaymneh L, Woessner JF: Neutral proteases capable of proteoglycan digesting activity in osteoarthritic and normal human articular cartilage. Arthritis Rheum. 1984, 27: 305-312.View ArticlePubMedGoogle Scholar
- Roughley PJ, White RJ, Poole AR: Identification of a hyaluronic acid-binding protein that interferes with the preparation of high-buoyant-density proteoglycan aggregates from adult human articular cartilage. Biochem J. 1985, 231: 129-138.PubMed CentralView ArticlePubMedGoogle Scholar
- Pelletier JP, Martel-Pelletier J, Cloutier JM, Woessner JF: Proteoglycan-degrading acid metalloprotease activity in human osteoarthritic cartilage, and the effect of intraarticular steroid injections. Arthritis Rheum. 1987, 30: 541-548.View ArticlePubMedGoogle Scholar
- Farndale RW, Sayers CA, Barrett AJ: A direct spectrophotometric microassay for sulfated glycosaminoglycans in cartilage cultures. Connect Tissue Res. 1982, 9: 247-248.View ArticlePubMedGoogle Scholar
- Billinghurst RC, Dahlberg L, Ionescu M, Reiner A, Bourne R, Rorabeck C, Mitchell P, Hambor J, Diekmann O, Tschesche H, et al: Enhanced cleavage of Type II collagen by collagenases in osteoarthritic articular cartilage. J Clin Invest. 1997, 99: 1534-1545.PubMed CentralView ArticlePubMedGoogle Scholar
- Knauper V, Lopez-Otin C, Smith B, Knight G, Murphy G: Biochemical characterization of human collagenase-3. J Biol Chem. 1996, 271: 1544-1550. 10.1074/jbc.271.3.1544.View ArticlePubMedGoogle Scholar
- Scott JE: Proteoglycan: collagen interactions in connective tissues. Ultrastructural, biochemical, functional and evolutionary aspects. Int J Biol Macromol. 1991, 13: 157-161. 10.1016/0141-8130(91)90041-R.View ArticlePubMedGoogle Scholar
- Nagase H: Activation mechanisms of matrix metalloproteinases. Biol Chem. 1997, 378: 151-160.PubMedGoogle Scholar
- Yamaguchi Y, Mann DM, Ruoslahti E: Negative regulation of transforming growth factor-beta by the proteoglycan decorin. Nature. 1990, 346: 281-284. 10.1038/346281a0.View ArticlePubMedGoogle Scholar
- Tardif G, Pelletier JP, Dupuis M, Geng C, Cloutier JM, Martel-Pelletier J: Collagenase 3 production by human osteoarthritic chondrocytes in response to growth factors and cytokines is a function of the physiological state of the cells. Arthritis Rheum. 1999, 42: 1147-1158. 10.1002/1529-0131(199906)42:6<1147::AID-ANR11>3.0.CO;2-Y.View ArticlePubMedGoogle Scholar
- Moldovan F, Pelletier JP, Hambor J, Cloutier JM, Martel-Pelletier J: Collagenase-3 (matrix metalloprotease 13) is preferentially localized in the deep layer of human arthritic cartilage in situ : In vitro mimicking effect by transforming growth factor beta. Arthritis Rheum. 1997, 40: 1653-1661.View ArticlePubMedGoogle Scholar
- Svensson L, Heinegard D, Oldberg A: Decorin-binding sites for collagen type I are mainly located in leucine-rich repeats 4–5. J Biol Chem. 1995, 270: 20712-20716. 10.1074/jbc.270.35.20712.View ArticlePubMedGoogle Scholar
- Scott PG, McEwan PA, Dodd CM, Bergmann EM, Bishop PN, Bella J: Crystal structure of the dimeric protein core of decorin, the archetypal small leucine-rich repeat proteoglycan. Proc Natl Acad Sci USA. 2004, 101: 15633-15638. 10.1073/pnas.0402976101.PubMed CentralView ArticlePubMedGoogle Scholar
- Kojoh K, Fukuda E, Matsuzawa H, Wakagi T: Zinc-coordination of aspartic acid-76 in Sulfolobus ferredoxin is not required for thermal stability of the molecule. J Inorg Biochem. 2002, 89: 69-73. 10.1016/S0162-0134(01)00410-X.View ArticlePubMedGoogle Scholar
- Reboul P, Pelletier JP, Tardif G, Cloutier JM, Martel-Pelletier J: The new collagenase, collagenase-3, is expressed and synthesized by human chondrocytes but not by synoviocytes: A role in osteoarthritis. J Clin Invest. 1996, 97: 2011-2019.PubMed CentralView ArticlePubMedGoogle Scholar
This article is published under license to BioMed Central Ltd. This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.