Analysis of ADAMTS4 and MT4-MMP indicates that both are involved in aggrecanolysis in interleukin-1-treated bovine cartilage

Patwari, Parth; Gao, Guangxia; Lee, Jennifer H.; Grodzinsky, Alan J.; Sandy, John D.

doi:10.1016/j.joca.2004.10.023

Cited by 74 publications

(89 citation statements)

References 49 publications

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“…ADAMTS-4 has been reported to be a potent aggrecan-degrading enzyme in articular cartilage (2,7,8,10,11,39,57), yet we detected only a moderate increase in this activity compared with that of ADAMTS-5 in mouse cartilage explants stimulated with IL-1␣ or retinoic acid. There are two main reasons for this result.…”

Section: Discussioncontrasting

confidence: 57%

Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

Ilic

East

Rogerson

et al. 2007

Journal of Biological Chemistry

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Aggrecan loss from mouse cartilage is predominantly because of ADAMTS-5 activity; however, the relative contribution of other proteolytic and nonproteolytic processes to this loss is not clear. This is the first study to compare aggrecan loss with aggrecan processing in mice with single and double deletions of ADAMTS-4 and -5 activity (⌬cat). Cartilage explants harvested from single and double ADAMTS-4 and -5 ⌬cat mice were cultured with or without interleukin (IL)-1␣ or retinoic acid and analyzed for (i) the kinetics of 35 S-labeled aggrecan loss, (ii) the pattern of 35 S-labeled aggrecan fragments released into the media and retained in the matrix, (iii) the pattern of total aggrecan fragments released into the media and retained in the matrix, and (iv) specific cleavage sites within the interglobular and chondroitin sulfate-2 domains. The loss of radiolabeled aggrecan from ADAMTS-4/-5 ⌬cat cartilage was less than that from ADAMTS-4, ADAMTS-5, or wild-type cartilage under nonstimulated conditions. IL-1␣ and retinoic acid stimulated radiolabeled aggrecan loss from wild-type and ADAMTS-4 ⌬cat cartilage, but there was little effect on ADAMTS-5 cartilage. Proteolysis of aggrecan contributed most to its loss in wild-type, ADAMTS-4, and ADAMTS-5 ⌬cat cartilage explants. The pattern of proteolytic processing of aggrecan in these cultures was consistent with that occurring in cartilage pathologies. Retinoic acid, but not IL-1␣, stimulated radiolabeled aggrecan loss from ADAMTS-4/-5 ⌬cat cartilage explants. Even though there was a 300% increase in aggrecan loss from ADAMTS-4/-5 ⌬cat cartilage stimulated with retinoic acid, the loss was not associated with aggrecanase cleavage but with the release of predominantly intact aggrecan consistent with the phenotype of the ADAMTS-4/-5 ⌬cat mouse. Our results show that chondrocytes have additional mechanism for the turnover of aggrecan and that when proteolytic mechanisms are blocked by ablation of aggrecanase activity, nonproteolytic mechanisms compensate to maintain cartilage homeostasis.Aggrecan catabolism has been studied extensively in the past 30 -40 years as part of the metabolic processes important in normal functional cartilage and in cartilage pathology. The major enzymes involved in degradation of aggrecan in normal and pathological cartilage are aggrecanases members of the ADAMTS 2 subfamily of adamalysin (M12) metalloproteinases (1, 2). These enzymes cleave the aggrecan core protein at aggrecanase-specific cleavage sites between glutamate and a small hydrophobic amino acid residue (3, 4). Most studies suggest that in vitro and in vivo ADAMTS-4 and -5 are the main aggrecanases (5-11); however, ADAMTS-1, -8, -9, -15, -16, and -18 also exhibit aggrecanase activity (12-15). Recombinant ADAMTS-4 and -5 have been suggested to be the most potent aggrecanases (15-19), although the relative potency in aggrecan degrading activity has yet to be clarified (11,20,21). In addition, recombinant ADAMTS-1, -4, and -5 cleave aggrecan at 5 sites: one within the interglobular domain a...

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Section: Discussioncontrasting

confidence: 57%

Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

Ilic

East

Rogerson

et al. 2007

Journal of Biological Chemistry

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“…We (13,14) and others (25,34) have previously reported that ADAMTS-4-1 exhibits little activity against the Glu 373 -Ala 374 bond in the IGD of aggrecan, whereas ADAMTS-4-2 is more effective at this site. On the other hand, Hashimoto et al (35) reported that full-length ADAMTS-4 was as active as a form of the enzyme lacking the spacer domain in cleaving the Glu 373 -Ala 374 bond, suggesting that removal of the spacer domain is not required for full catalytic activity.…”

Section: Discussionmentioning

confidence: 66%

Functional Differences of the Catalytic and Non-catalytic Domains in Human ADAMTS-4 and ADAMTS-5 in Aggrecanolytic Activity

Fushimi

Troeberg

Nakamura

et al. 2008

Journal of Biological Chemistry

106

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ADAMTS-4 (aggrecanase-1) and ADAMTS-5 (aggrecanase-2) are multidomain metalloproteinases belonging to the ADAMTS family. We have previously reported that human ADAMTS-5 has much higher aggrecanolytic activity than human ADAMTS-4. To investigate the different proteolytic activity of the two enzymes, we generated a series of chimeras by exchanging various non-catalytic domains of the two proteinases. We found that the catalytic domain of ADAMTS-5 has higher intrinsic catalytic ability than that of ADAMTS-4. The studies also demonstrated that the non-catalytic domains of ADAMTS-5 are more effective modifiers than those of ADAMTS-4, making both catalytic domains more active against aggrecan, an A common feature of osteoarthritis (OA) 2 and rheumatoid arthritis is destruction of articular cartilage, which is characterized by a homeostatic imbalance between synthesis and degradation of the extracellular matrix (ECM). The destructive process is believed to be due to elevated activities of proteolytic enzymes that degrade macromolecules of the cartilage ECM such as aggrecan and type II collagen fibrils. Aggrecan is the major proteoglycan in articular cartilage, and it forms large aggregates by interacting with hyaluronan and link protein.Aggrecan monomers consist of a core protein with chondroitin sulfate (CS) and keratan sulfate (KS) polysaccharide chains. The core protein consists of several segments, including an N-terminal globular domain (G1), an interglobular domain (IGD), a second globular domain (G2), a long glycosaminoglycan (GAG) attachment region, including KS-rich and CS-rich (CS-1 and CS-2) regions, and a C-terminal globular domain (G3) (1). Aggrecans are highly hydrated because of their long negatively charged polysaccharide chains. Thus, within the collagen framework, they enable the cartilage to resist mechanical compression as a load-bearing surface. Aggrecan loss is therefore considered to be a crucial initial event in the development of arthritis, which is followed by essentially irreversible collagen degradation (2, 3). This pathological aggrecan degradation in articular cartilage is driven mainly by proteolytic enzymes termed aggrecanases and matrix metalloproteinases (MMPs) (4, 5).The aggrecanases are members of the ADAMTSs (a disintegrin and metalloproteinase with thrombospondin type 1 motifs) family (6). The first two proteinases identified to cleave the aggrecan core protein at the aggrecanase-specific Glu 373 -Ala 374 bond in the IGD were ADAMTS-4 (aggrecanase-1) (7) and ADAMTS-5 (aggrecanase-2) (8). Finding aggrecan fragments cleaved at the Glu 373 -Ala 374 bond in synovial fluids and cartilage from patients with OA and rheumatoid arthritis suggested that aggrecanases play an important role in cartilage destruction (9, 10). They are also the primary enzymes that cleave aggrecan in response to inflammatory cytokines in articular cartilage explant systems (11). Gendron et al. (12) have shown that cleavage of the Glu 373 -Ala 374 bond in interleukin-1-stimulated cartilage explants is blocked by ...

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“…The less profound effects seen with the MMPinhibitor may have been due to an inhibition of MMP-mediated activation of aggrecanases. A mechanism by which MMP-17 cleaves ADAMTS-4 at the cell surface appears to be active in IL-1-stimulated bovine cartilage explants (Patwari, et al, 2005), and inhibition of this mechanism with the MMP-selective inhibitor may have resulted in reduced levels of fully activated enzyme Gao, et al, 2004). The MMP-and nonselective inhibitors prevented IL-1-induced generation of the VDIPES neoepitope, indicating that MMPs also degrade aggrecan in this model.…”

Section: Discussionmentioning

confidence: 76%

“…Proinflammatory stimuli may upregulate transcription of aggrecanase genes (Bau, et al, 2002;Koshy, et al, 2002), and there is mounting evidence for substantial post-translational processing of the enzymes that alter aggrecanase activity and specificity (Pratta, et al, 2003a). In human chondrosarcoma cells and bovine cartilage explants, MMP-17 (MT4-MMP) appears to be responsible for C-terminal truncation of ADAMTS-4, a process which converts the enzyme from one which can cleave only the sGAG-rich region to one which can also cleave the interglobular domain (IGD) (Gao, et al, 2004;Patwari, et al, 2005). Aggrecanase activity within the IGD is marked by scission of the NITEGE 392 -393 ARGSVI bond (Sandy, et al, 1991).…”

Section: Introductionmentioning

confidence: 99%

Selective and non-selective metalloproteinase inhibitors reduce IL-1-induced cartilage degradation and loss of mechanical properties

et al. 2007

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Articular cartilage undergoes matrix degradation and loss of mechanical properties when stimulated with proinflammatory cytokines such as interleukin-1 (IL-1). Aggrecanases and matrix metalloproteinases (MMPs) are thought to be principal downstream effectors of cytokine-induced matrix catabolism, and aggrecanase-or MMP-selective inhibitors reduce or block matrix destruction in several model systems. The objective of this study was to use metalloproteinase inhibitors to perturb IL-1-induced matrix catabolism in bovine cartilage explants and examine their effects on changes in tissue compression and shear properties. Explanted tissue was stimulated with IL-1 for up to 24 days in the absence or presence of inhibitors which were aggrecanase-selective, MMP-selective, or non-selective. Analysis of conditioned media and explant digests revealed that aggrecanase-mediated aggrecanolysis was delayed to varying extents with all inhibitor treatments, but that aggrecan release persisted. Collagen degradation was abrogated by MMP-and non-selective inhibitors and reduced by the aggrecanase inhibitor. The inhibitors delayed but did not reduce loss of the equilibrium compression modulus, whereas the loss of dynamic compression and shear moduli was delayed and reduced. The data suggest that non-metalloproteinase mechanisms participate in IL-1-induced matrix degradation and loss of tissue material properties.

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Analysis of ADAMTS4 and MT4-MMP indicates that both are involved in aggrecanolysis in interleukin-1-treated bovine cartilage

Cited by 74 publications

References 49 publications

Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

Distinguishing Aggrecan Loss from Aggrecan Proteolysis in ADAMTS-4 and ADAMTS-5 Single and Double Deficient Mice

Functional Differences of the Catalytic and Non-catalytic Domains in Human ADAMTS-4 and ADAMTS-5 in Aggrecanolytic Activity

Selective and non-selective metalloproteinase inhibitors reduce IL-1-induced cartilage degradation and loss of mechanical properties

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