A high throughput enzymatic assay for stromelysin-1

Pratta, Michael A.; Blessington, D. L.; Trzaskos, James M.; Arner, Elizabeth C.

doi:10.1007/s000110050134

Cited by 2 publications

(2 citation statements)

References 5 publications

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“…Thus, this region may serve to bind aggrecanase-1 to the glycosaminoglycans of the aggrecan substrate. Deglycosylation of aggrecan decreases the ability of aggrecanase to generate fragments formed by cleavage at the G 1~~~~-A l a~~~ bond (19), which is consistent with this hypothesis.…”

supporting

confidence: 58%

Purification and Cloning of Aggrecanase-1: A Member of the ADAMTS Family of Proteins

Tortorella¹,

Burn²,

Pratta³

et al. 1999

Science

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658

476

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We purified, cloned, and expressed aggrecanase, a protease that is thought to be responsible for the degradation of cartilage aggrecan in arthritic diseases. Aggrecanase-1 [a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4)] is a member of the ADAMTS protein family that cleaves aggrecan at the glutamic acid-373-alanine-374 bond. The identification of this protease provides a specific target for the development of therapeutics to prevent cartilage degradation in arthritis.

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supporting

confidence: 58%

Purification and Cloning of Aggrecanase-1: A Member of the ADAMTS Family of Proteins

Tortorella¹,

Burn²,

Pratta³

et al. 1999

Science

Self Cite

658

476

View full text Add to dashboard Cite

show abstract

“…The TSP motif and submotif of ADAMTS11 may be responsible for binding to the glycosaminoglycans of aggrecan. Consistent with this hypothesis is the observation that deglycosylation of aggrecan decreases the production of Glu 373 -Ala 374 cleavage products by aggrecanases (58). Sequences within the spacer region between the TSP motifs of ADAMTS11 are conserved in all the family members and are likely to provide an additional adhesion motif, since recent studies demonstrate that sequences within the spacer region of murine ADAMTS1 can themselves bind tightly to the extracellular matrix (19).…”

Section: Fig 3 Expression Levels Of the Aggrecanases And Human Adammentioning

confidence: 81%

Cloning and Characterization of ADAMTS11 , an Aggrecanase from the ADAMTS Family

Abbaszade¹,

Liu²,

Yang³

et al. 1999

Journal of Biological Chemistry

469

325

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Aggrecan is responsible for the mechanical properties of cartilage. One of the earliest changes observed in arthritis is the depletion of cartilage aggrecan due to increased proteolytic cleavage within the interglobular domain. Two major sites of cleavage have been identified in this region at Asn 341 -Phe 342 and Glu 373 -Ala 374 . While several matrix metalloproteinases have been shown to cleave at Asn 341 -Phe 342 , an as yet unidentified protein termed "aggrecanase" is responsible for cleavage at Glu 373 -Ala 374 and is hypothesized to play a pivotal role in cartilage damage. We have identified and cloned a novel disintegrin metalloproteinase with thrombospondin motifs that possesses aggrecanase activity, ADAMTS11 (aggrecanase-2), which has extensive homology to ADAMTS4 (aggrecanase-1) and the inflammationassociated gene ADAMTS1. ADAMTS11 possesses a number of conserved domains that have been shown to play a role in integrin binding, cell-cell interactions, and extracellular matrix binding. We have expressed recombinant human ADAMTS11 in insect cells and shown that it cleaves aggrecan at the Glu 373 -Ala 374 site, with the cleavage pattern and inhibitor profile being indistinguishable from that observed with native aggrecanase. A comparison of the structure and expression patterns of ADAMTS11, ADAMTS4, and ADAMTS1 is also described. Our findings will facilitate the study of the mechanisms of cartilage degradation and provide targets to search for effective inhibitors of cartilage depletion in arthritic disease.Aggrecan is the major proteoglycan of cartilage and is responsible for its compressibility and stiffness. Aggrecan contains two N-terminal globular domains, G 1 and G 2 , separated by a proteolyticaly sensitive interglobular domain, followed by a glycosaminoglycan attachment region and a C-terminal globular domain (G 3 ). The G 1 domain of aggrecan interacts with hyaluronic acid and link protein to form large aggregates containing multiple aggrecan monomers that are trapped within the cartilage matrix. Cleavage of aggrecan has been shown to occur at Asn 341 -Phe 342 and Glu 373 -Ala 374 within the interglobular domain, with the cleaved aggrecan being free to exit the matrix since it lacks the G 1 domain, which is responsible for formation of the high molecular weight complexes. Results from several studies suggest that cleavage at the Glu 373 -Ala 374 site is responsible for the increased aggrecan degradation observed in inflammatory joint disease. Products resulting from cleavage at the Glu 373 -Ala 374 site have been shown to accumulate in cartilage explants and chondrocyte cultures treated with interleukin-1 and retinoic acid (1-5) and in the synovial fluid of patients with osteoarthritis and inflammatory joint disease (6, 7). While several characterized matrix metalloproteases 1 have been shown to cleave at the Asn 341 -Phe 342 site (8 -14), they are not responsible for the observed cleavage at Glu 373 -Ala 374 . A novel proteolytic activity, termed "aggrecanase," has been hypothesized to be respo...

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A high throughput enzymatic assay for stromelysin-1

Cited by 2 publications

References 5 publications

Purification and Cloning of Aggrecanase-1: A Member of the ADAMTS Family of Proteins

Purification and Cloning of Aggrecanase-1: A Member of the ADAMTS Family of Proteins

Cloning and Characterization of ADAMTS11 , an Aggrecanase from the ADAMTS Family

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