Proteolytic Activities of Human ADAMTS-5

Gendron, Christi M.; Kashiwagi, Masahide; Lim, Ngee Han; Enghild, Jan J.; Thøgersen, Ida B.; Hughes, Claire E.; Caterson, Bruce; Nagase, Hideaki

doi:10.1074/jbc.m701523200

Cited by 226 publications

(153 citation statements)

References 52 publications

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“…Our recent studies showed that the aggrecanolytic activity of ADAMTS-5 was much greater than ADAMTS-4 in both the IGD and the CS-2 regions (13). This provided biochemical evidence supporting the view that ADAMTS-5 is the major aggrecanase in cartilage catabolism and pathology.…”

supporting

confidence: 56%

“…The rabbit anti-GELE antibody that recognizes the new GELE 1480 C terminus generated by aggrecanase cleavage of the aggrecan core protein has been previously described (14). The rabbit anti-human ADAMTS-4 catalytic domain and anti-human ADAMTS-5 catalytic domain antibodies were raised in rabbits and purified as described previously (13,14). All other reagents used were of the highest analytical grade available.…”

Section: Methodsmentioning

confidence: 99%

“…1). We have previously shown that the non-catalytic ancillary domains of ADAMTS-4 and -5 play a major role in regulating aggrecanase activity and substrate specificity (13,14). Furthermore, Flannery et al (23) showed that the CysR and Sp domains of ADAMTS-4 are necessary for the enzyme's interaction with the GAG chains of aggrecan, and Tortorella et al (24) reported that the TS domain of ADAMTS-4 is critical for aggrecan recognition and cleavage.…”

mentioning

confidence: 99%

“…Among the ADAMTS enzymes, ADAMTS-4 and -5 have received most attention in the pathology of arthritic joint diseases, because they are the most efficient aggrecanases in vitro (13,14). Other ADAMTSs, such as ADAMTS-1, -8, -9, -15, -16, and -18 have been shown to cleave aggrecan, but they have very weak activity (6,15).…”

mentioning

confidence: 99%

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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

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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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supporting

confidence: 56%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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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

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106

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“…Unexpectedly, however, no detectable aggrecanase-1 mRNA expression was found in chondrocytes cultured with T-2 toxin in vitro. Previous studies have shown that aggrecanase-1 usually is an up-regulated expression in the cartilage induced by IL-1 (Liacini et al, 2005;Tsuzaki et al, 2003;Pratta et al, 2003), whereas aggrecanse-2 mRNA is expressed constitutively in cartilage, and the aggrecanase activity of aggrecanase-2 was at least 1 000-fold greater than that of aggrecanase-1 (Gendron et al, 2007). Therefore, aggrecanase-2 is a major aggrecanase in cartilage metabolism and pathology.…”

Section: Discussionmentioning

confidence: 97%

Promotion of the articular cartilage proteoglycan degradation by T-2 toxin and selenium protective effect

Liu

Cao

Shi

et al. 2008

J. Zhejiang Univ. Sci. B

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Abstract:Objective: To identify the relationship between T-2 toxin and Kashin-Beck disease (KBD), the effects of T-2 toxin on aggrecan metabolism in human chondrocytes and cartilage were investigated in vitro. Methods: Chondrocytes were isolated from human articular cartilage and cultured in vitro. Hyaluronic acid (HA), soluble CD44 (sCD44), IL-1β and TNF-α levels in supernatants were measured by enzyme-linked immunosorbent assay (ELISA). CD44 content in chondrocyte membrane was determined by flow cytometry (FCM). CD44, hyaluronic acid synthetase-2 (HAS-2) and aggrecanases mRNA levels in chondrocytes were determined using reverse transcription polymerase chain reaction (RT-PCR). Immunocytochemical method was used to investigate expressions of BC-13, 3-B-3(−) and 2-B-6 epitopes in the cartilage reconstructed in vitro. Results: T-2 toxin inhibited CD44, HAS-2, and aggrecan mRNA expressions, but promoted aggrecanase-2 mRNA expression. Meanwhile, CD44 expression was found to be the lowest in the chondrocytes cultured with T-2 toxin and the highest in control plus selenium group. In addition, ELISA results indicated that there were higher sCD44, IL-1β and TNF-α levels in T-2 toxin group. Similarly, higher HA levels were also observed in T-2 toxin group using radioimmunoprecipitation assay (RIPA). Furthermore, using monoclonal antibodies BC-13, 3-B-3 and 2-B-6, strong positive immunostaining was found in the reconstructed cartilage cultured with T-2 toxin, whereas no positive staining or very weak staining was observed in the cartilage cultured without T-2 toxin. Selenium could partly inhibit the effects of T-2 toxin above. Conclusion: T-2 toxin could inhibit aggrecan synthesis, promote aggrecanases and pro-inflammatory cytokines production, and consequently induce aggrecan degradation in chondrocytes. These will perturb metabolism balance between aggrecan synthesis and degradation in cartilage, inducing aggrecan loss in the end, which may be the initiation of the cartilage degradation.

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Genetic variation including nonsynonymous polymorphisms of a major aggrecanase, ADAMTS‐5, in susceptibility to osteoarthritis

Rodriguez‐Lopez

Mustafa

Pombo‐Suárez

et al. 2008

Arthritis & Rheumatism

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Objective. Given the recent characterization of ADAMTS-5 as the main aggrecanase of cartilage destruction in mouse models, we explored whether genetic variation and, in particular, putative damaging polymorphisms in the ADAMTS-5 gene modify susceptibility to osteoarthritis (OA).Methods. Two likely deleterious nonsynonymous single-nucleotide polymorphisms (SNPs) were identified in ADAMTS-5 by bioinformatics analysis, rs2830585 in exon 5 affecting a thrombospondin 1 motif, and rs226794 in exon 7. Exploration of their role was carried out in 3 steps, discovery, extension, and replication, on samples obtained from 4 European Caucasian collections, comprising a total of 2,715 patients with knee, hip, or hand OA and 1,185 OA-free controls. In addition, 6 tagSNPs were studied to fully evaluate genetic variation in the ADAMTS-5 locus.Results. Initial analyses of 2 sample collections (n ‫؍‬ 277 and n ‫؍‬ 159) showed a trend toward decreased frequency of the putative deleterious allele of rs226794 among patients with severe knee OA (P ‫؍‬ 0.047 versus controls). However, results in patients with knee OA from 2 additional sample collections (n ‫؍‬ 360 and n ‫؍‬ 265) did not confirm this trend. No association was found with hip OA or hand OA. None of the other SNPs or haplotypes constructed with these SNPs showed a significant association with OA susceptibility.Conclusion. Use of several collections of OA samples allowed us to obtain sound evidence against the participation of genetic variation in ADAMTS-5 in OA susceptibility. These results indicate the need to further explore the function of this aggrecanase in human OA to determine whether it is as critical as has been observed in mouse models.

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Proteolytic Activities of Human ADAMTS-5

Cited by 226 publications

References 52 publications

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

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

Promotion of the articular cartilage proteoglycan degradation by T-2 toxin and selenium protective effect

Genetic variation including nonsynonymous polymorphisms of a major aggrecanase, ADAMTS‐5, in susceptibility to osteoarthritis

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