Proteolytic Activities of Human ADAMTS-5

Abstract

Aggrecanases have been characterized as proteinases that cleave the Glu373-Ala374 bond of the aggrecan core protein, and they are multidomain metalloproteinases belonging to the ADAMTS (adamalysin with thrombospondin type 1 motifs) family. The first aggrecanases discovered were ADAMTS-4 (aggrecanase 1) and ADAMTS-5 (aggrecanase 2). They contain a zinc catalytic domain followed by non-catalytic ancillary domains, including a disintegrin domain, a thrombospondin domain, a cysteine-rich domain, and a spacer domain. In the case of ADAMTS-5, a second thrombospondin domain follows the spacer domain. We previously reported that the non-catalytic domains of ADAMTS-4 influence both its extracellular matrix interaction and proteolytic abilities. Here we report the effects of these domains of ADAMTS-5 on the extracellular matrix interaction and proteolytic activities and compare them with those of ADAMTS-4. Although the spacer domain was critical for ADAMTS-4 localization in the matrix, the cysteine-rich domain influenced ADAMTS-5 localization. Similar to previous reports of other ADAMTS family members, very little proteolytic activity was detected with the ADAMTS-5 catalytic domain alone. The sequential inclusion of each carboxyl-terminal domain enhanced its activity against aggrecan, carboxymethylated transferrin, fibromodulin, decorin, biglycan, and fibronectin. Both ADAMTS-4 and -5 had a broad optimal activity at pH 7.0–9.5. Aggrecanolytic activities were sensitive to the NaCl concentration, but activities on non-aggrecan substrates, e.g. carboxymethylated transferrin, were not affected. Although ADAMTS-4 and ADAMTS-5 had similar general proteolytic activities, the aggrecanase activity of ADAMTS-5 was at least 1,000-fold greater than that of ADAMTS-4 under physiological conditions. Our studies suggest that ADAMTS-5 is a major aggrecanase in cartilage metabolism and pathology. Aggrecanases have been characterized as proteinases that cleave the Glu373-Ala374 bond of the aggrecan core protein, and they are multidomain metalloproteinases belonging to the ADAMTS (adamalysin with thrombospondin type 1 motifs) family. The first aggrecanases discovered were ADAMTS-4 (aggrecanase 1) and ADAMTS-5 (aggrecanase 2). They contain a zinc catalytic domain followed by non-catalytic ancillary domains, including a disintegrin domain, a thrombospondin domain, a cysteine-rich domain, and a spacer domain. In the case of ADAMTS-5, a second thrombospondin domain follows the spacer domain. We previously reported that the non-catalytic domains of ADAMTS-4 influence both its extracellular matrix interaction and proteolytic abilities. Here we report the effects of these domains of ADAMTS-5 on the extracellular matrix interaction and proteolytic activities and compare them with those of ADAMTS-4. Although the spacer domain was critical for ADAMTS-4 localization in the matrix, the cysteine-rich domain influenced ADAMTS-5 localization. Similar to previous reports of other ADAMTS family members, very little proteolytic activity was detected with the ADAMTS-5 catalytic domain alone. The sequential inclusion of each carboxyl-terminal domain enhanced its activity against aggrecan, carboxymethylated transferrin, fibromodulin, decorin, biglycan, and fibronectin. Both ADAMTS-4 and -5 had a broad optimal activity at pH 7.0–9.5. Aggrecanolytic activities were sensitive to the NaCl concentration, but activities on non-aggrecan substrates, e.g. carboxymethylated transferrin, were not affected. Although ADAMTS-4 and ADAMTS-5 had similar general proteolytic activities, the aggrecanase activity of ADAMTS-5 was at least 1,000-fold greater than that of ADAMTS-4 under physiological conditions. Our studies suggest that ADAMTS-5 is a major aggrecanase in cartilage metabolism and pathology. Destruction of articular cartilage is a feature of various arthritides, including rheumatoid and osteoarthritis, that results in joint impairment and disability. It is caused primarily by an elevation in proteolytic enzymes that degrade macromolecules of the cartilage extracellular matrix. Aggrecan degradation is initially observed followed by essentially irreversible collagen degradation. The proteinases that are responsible for aggrecan degradation in cartilage are matrix metalloproteinases (MMPs) 4The abbreviations used are: MMP, matrix metalloproteinase; ADAMTS, a disintegrin and metalloproteinase with thrombospondin type 1 motifs; anti-TS5cat, rabbit anti-ADAMTS-5 catalytic domain; Cm-Tf, reduced, carboxymethylated transferrin; CysR, cysteine-rich; Dis, disintegrin; DMEM, Dulbecco's modified Eagle's medium; IGD, interglobular domain of aggrecan; IL-1, interleukin-1; MT-MMP, membrane-type matrix metalloproteinase; N-TIMP, amino-terminal domain of tissue inhibitor of metalloproteinases; Sp, spacer; TIMP, tissue inhibitor of metalloproteinases; TS, thrombospondin type 1; MES, 4-morpholineethanesulfonic acid; ECM, extracellular matrix. and “aggrecanases,” members of the ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 motifs) family (1Nagase H. Kashiwagi M. Arthritis Res. Ther. 2003; 5: 94-103Crossref PubMed Google Scholar, 2Porter S. Clark I.M. Kevorkian L. Edwards D.R. Biochem. J. 2005; 386: 15-27Crossref PubMed Scopus (625) Google Scholar). Aggrecanase activity was first defined as the ability to cleave the Glu373-Ala374 bond in the interglobular domain (IGD) of the aggrecan core protein (3Sandy J.D. Neame P.J. Boynton R.E. Flannery C.R. J. Biol. Chem. 1991; 266: 8683-8685Abstract Full Text PDF PubMed Google Scholar, 4Ilic M.Z. Handley C.J. Robinson H.C. Mok M.T. Arch. Biochem. Biophys. 1992; 294: 115-122Crossref PubMed Scopus (164) Google Scholar). The first two proteinases shown to be capable of cleaving aggrecan at this site were ADAMTS-4 (aggrecanase 1) (5Tortorella M.D. Burn T.C. Pratta M.A. Abbaszade I. Hollis J.M. Liu R. Rosenfeld S.A. Copeland R.A. Decicco C.P. Wynn R. Rockwell A. Yang F. Duke J.L. Solomon K. George H. Bruckner R. Nagase H. Itoh Y. Ellis D.M. Ross H. Wiswall B.H. Murphy K. Hillman Jr., M.C. Hollis G.F. Newton R.C. Magolda R.L. Trzaskos J.M. Arner E.C. Science. 1999; 284: 1664-1666Crossref PubMed Scopus (625) Google Scholar), and ADAMTS-5 5ADAMTS-5 was originally referred to as “ADAMTS-11.” (aggrecanase 2) (6Abbaszade I. Liu R.Q. Yang F. Rosenfeld S.A. Ross O.H. Link J.R. Ellis D.M. Tortorella M.D. Pratta M.A. Hollis J.M. Wynn R. Duke J.L. George H.J. Hillman Jr., M.C. Murphy K. Wiswall B.H. Copeland R.A. Decicco C.P. Bruckner R. Nagase H. Itoh Y. Newton R.C. Magolda R.L. Trzaskos J.M. Hollis G.F. Arner E.C. Burn T.C. J. Biol. Chem. 1999; 274: 23443-23450Abstract Full Text Full Text PDF PubMed Scopus (447) Google Scholar). More recently, ADAMTS-1, -8, -9, -15, -16, and -18 were shown to cleave the Glu373-Ala374 bond in the IGD at a high enzyme-substrate ratio (2Porter S. Clark I.M. Kevorkian L. Edwards D.R. Biochem. J. 2005; 386: 15-27Crossref PubMed Scopus (625) Google Scholar, 7Zeng W. Corcoran C. Collins-Racie L.A. LaVallie E.R. Morris E.A. Flannery C.R. Biochim. Biophys. Acta. 2006; 1760: 517-524Crossref PubMed Scopus (91) Google Scholar). Although this bond is also cleaved by MMP-8 (8Fosang A.J. Last K. Neame P.J. Murphy H. Biochem. J. PubMed Scopus Google and A. H. Biochem. J. PubMed Scopus Google at high in the site in the IGD is to be at the bond (1Nagase H. Kashiwagi M. Arthritis Res. Ther. 2003; 5: 94-103Crossref PubMed Google Scholar). The of the Glu373-Ala374 bond in articular cartilage is by tissue inhibitor of metalloproteinases but not by other C. Kashiwagi M. C. Nagase H. 2003; PubMed Scopus Google Scholar). is a of ADAMTS metalloproteinases as the proteinases responsible for the observed aggrecanase the that have aggrecanase ADAMTS-4 than other the elevation of its is observed in with IL-1, ADAMTS-5 is not M.D. C. Arner Full Text PDF PubMed Scopus Google Scholar, J. Arthritis PubMed Scopus Google Scholar). not on cartilage aggrecan with the in an by joint R. Flannery C.R. K. Yang Morris E.A. Arthritis PubMed Scopus Google Scholar). were not against joint in an L. M.A. A.J. Arthritis 2005; PubMed Scopus Google Scholar). studies suggest that ADAMTS-4 be the major that aggrecan the of the other studies with that the of ADAMTS-5 against cartilage R. Flannery C.R. K. Yang Morris E.A. 2005; PubMed Scopus Google H. C.J. Last K. P.J. A.J. 2005; PubMed Scopus Google was that ADAMTS-5 a in aggrecan degradation at least in these of in ADAMTS-4 and ADAMTS-5 are multidomain metalloproteinases the the extracellular Both enzymes have a similar domain of a a catalytic metalloproteinase domain, a disintegrin domain, a thrombospondin type domain, a cysteine-rich domain, and a spacer domain. In ADAMTS-5 an domain the spacer domain 1 for domain We have shown previously that the non-catalytic ancillary domains of ADAMTS-4 a major in aggrecanase activity M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). In ADAMTS-4 the aggrecan core protein but little activity to cleave the initially characterized aggrecanase Glu373-Ala374 M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, A. S. F. J.D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), other non-aggrecan protein M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the carboxyl-terminal domain is the activity for the Glu373-Ala374 bond as as proteolytic activities against as reduced, carboxymethylated fibromodulin, and M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). ADAMTS-4 is to a similar to that of by in A. S. F. J.D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google and cartilage A.J. J.D. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). that the of the ADAMTS-4 spacer domain and ADAMTS-4 proteolytic in little proteolytic activity was detected with the metalloproteinase domain M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the of ADAMTS-5 is The studies its ability to degrade aggrecan, and W. Corcoran C. Collins-Racie L.A. LaVallie E.R. Morris E.A. Flannery C.R. Biochim. Biophys. Acta. 2006; 1760: 517-524Crossref PubMed Scopus (91) Google Scholar, I. M.Z. Handley C.J. Clark M. Liu C. R.A. J. Biochem. PubMed Scopus Google Scholar, M.D. Liu R.Q. Burn Newton R.C. Arner Biol. PubMed Scopus Google Scholar, M.D. Arner E.C. R. A. J. K. A. Liu R.Q. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, J. C. R. J. 2006; PubMed Scopus Google Scholar). We the of the non-catalytic domains of ADAMTS-5 by enzymes we have the ancillary domains the We activities for the aggrecan core protein, Cm-Tf, and other cartilage matrix studies that ADAMTS-5 is to cleave aggrecan core protein than ADAMTS-4. this that the of the non-catalytic domains for activities and the the two enzymes are Our results to that ADAMTS-5 is the major and the were and were and and for were and and were biglycan, fibromodulin, and were enzymes were and were and were The was Science. amino-terminal domain of tissue inhibitor of and were as previously C. Kashiwagi M. C. Nagase H. 2003; PubMed Scopus Google Scholar). Aggrecan was cartilage to the of and J. Biol. Chem. Full Text PDF PubMed Google Scholar). was by type collagen Y. S. Nagase H. Biochem. J. PubMed Scopus Google Scholar). was as previously Y. PubMed Scopus Google Scholar). that the amino-terminal of the aggrecan core protein and that the of aggrecan core protein were as previously J.R. J.R. Google Scholar, A.J. P.J. Biochem. J. PubMed Scopus Google Scholar). The rabbit that the by aggrecanase of the aggrecan core protein was in a rabbit against the aggrecan in are not of the aggrecan but for the of the to The rabbit anti-ADAMTS-5 catalytic domain was in a rabbit against the and to Both were by of for ADAMTS-5 and the ADAMTS-5 was the of and ADAMTS-5 and its domain were the The was with as a and by two a site and a a site a and the in The was for of at at and at The were the the to the and both the and the and that were the ADAMTS-5 at the of the ADAMTS-5 to is shown in and of the ADAMTS-5 and ADAMTS-5 its domain was with The were for in DMEM, and a of the protein, the was with and In the case of and was the to in the the were and at was not for this was The were to and a at The was first with 1 and The was with in and and were on a to the to the of in the of a was used to the and to the were against and to a of The was with and was the with 1 and The were against aggrecanase and and at of ADAMTS-5 and of ADAMTS-5 and its domain were by with The of and are the catalytic domain was with as to of the were at of a in The were and the with of to the The the were the were with to and of with was to each and for was The were and were by at for in the were a of and to with the to ADAMTS-5 and its of ADAMTS-5 in were essentially the as those for ADAMTS-4 studies M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). were with the ADAMTS-5 its and in with for each ADAMTS-5 protein, the were with in for and with The were with the followed by The were with with in and with to the The were a with a with and at and for and at and for and was under a of the of M. A. Biochem. PubMed Scopus Google Scholar). were with with A. M. M. Chem. PubMed Scopus Google Scholar). were a and the was as previously M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). Aggrecanase was with ADAMTS-5, an ADAMTS-5 domain in of aggrecanase and at for the of The were by an of and Aggrecan was by with of of aggrecan and of of aggrecan for at The were by of at for and at for The was and in of The were by with the as previously M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). the were and were under conditions. activities of ADAMTS-5 and were by of for of on the of the and the of and were a and the was the of and was with in aggrecanase at for and the were by The were to a and with The were and a and were by degradation in an with high was with for and the were with to The was in a similar to that for the ADAMTS-5 and its carboxyl-terminal were the of by The used to the was by of the a by with as two to The were to be ADAMTS-5 by of of of of of and of were 1 of of ADAMTS-5 were but was observed and The of the enzymes by were than that with of the protein W. Corcoran C. Collins-Racie L.A. LaVallie E.R. Morris E.A. Flannery C.R. Biochim. Biophys. Acta. 2006; 1760: 517-524Crossref PubMed Scopus (91) Google Scholar). ADAMTS-5 and The were in the in the domain, in the domain, and two in the domain. site was in the domain. these not the catalytic domain at a than was as two that were both by the and The of this with in a of that the is an of of in with the that was with the and The of to the the of that to on the and in the Both the carboxyl-terminal domain and the and domain were also to the ECM, but of the domain the the suggest that the domain of ADAMTS-5 a in studies were with of ADAMTS-4 and ADAMTS-5 as by site with The is proteolytic activity was the of was by with as the carboxyl-terminal domain, was used for the with ADAMTS-5 this of ADAMTS-4 the of general activity as with ADAMTS-4. of the general aggrecanase activity the activity against the Glu373-Ala374 bond of the aggrecan Aggrecan activities of the various of ADAMTS-5 were by the the that the of the aggrecan core protein (a of general aggrecanase the that the amino-terminal aggrecanase activity in the IGD of aggrecan and the that the carboxyl-terminal aggrecanase activity in the of aggrecan Aggrecanolytic activity to as was each and the results are shown in proteolytic activities of ADAMTS-4 and in a with the the general of aggrecan degradation by ADAMTS-5 and its domain of the that and had the general aggrecanase activity with activity was by the of the domain and by the of the and the first domains Aggrecan degradation was detected with the catalytic domain It is that the aggrecan of is that of that the and domains influence both and of ADAMTS-4 and -5 that ADAMTS-5 was than ADAMTS-4. In ADAMTS-4 an that was not in the ADAMTS-5 that ADAMTS-4 aggrecan at a the that is in cleaving the Glu373-Ala374 bond in the IGD of the carboxyl-terminal domain the activity by be observed with of the domain we the of to be similar to the of of the domain caused a in was than activity was detected 1 a domain similar to that of was than at this the were with the and the activity and cleaved the bond to a similar activity to with that the domain is for the at this The activity was of the domain. activity was detected with and under the with ADAMTS-4 that was than at this site The aggrecanase activity of ADAMTS-4 is aggrecan is M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, M. Pratta M. Liu R.Q. Abbaszade I. Ross H. Burn Arner J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). We and aggrecan with and is shown in the aggrecanase activity of ADAMTS-4 and -5 at the site was the aggrecan was Similar results were the were the not was at least than on against ADAMTS-4 little general proteolytic but of the domain the activity against M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). ADAMTS-5 and to a similar and that of the carboxyl-terminal domain and the domain not its The of the domain caused in this in activity was with the of the domain but very little activity at as high as of and that was on of the activity by but not by that this activity is to the of that ADAMTS-5 and its domain have similar but they were those of ADAMTS-4 that the two enzymes is not of the by that of its were also cleaved by but that ADAMTS-5 a of in ADAMTS-5 at major and and in a similar to that of The of the domain the domain and the domain the ability of to cleave results were similar to those observed for with a of ADAMTS-4 ADAMTS-5 catalytic domain both enzymes have very activity the ADAMTS-5 catalytic domain was than the ADAMTS-4 catalytic domain The of the by ADAMTS-5 was that at the an site cleaved by ADAMTS-4 M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google and L. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus (91) Google Scholar). and biglycan, were also by ADAMTS-4 and -5 that was a for was for ADAMTS-5 Both enzymes but at a very of ADAMTS-5, with the of also not In both ADAMTS-4 and -5 but activity was detected not The activities of ADAMTS-4 and ADAMTS-5 were aggrecan and as Both enzymes a broad optimal activity against aggrecan pH and similar pH optimal activity was detected was used as and We that the of the aggrecanase activity by the interaction the and the of the aggrecan this we aggrecan with ADAMTS-4 ADAMTS-5 in various of shown in the aggrecanase activity of was by the of The activity was detected with but the was of the activity was essentially activity was detected at The aggrecanase activity of ADAMTS-5 was also by NaCl but the on ADAMTS-4. activity was very at NaCl aggrecanase activity for ADAMTS-5 was detected and of its activity was at In to aggrecanase the general proteolytic activities of ADAMTS-4 and as with Cm-Tf, were by to 1 NaCl the results to results that the interaction the ancillary domains of the two and of aggrecan activities, general proteolytic activities are of that the proteolytic activities of multidomain ADAMTS metalloproteinases on are by non-catalytic ancillary the activity of is by its carboxyl-terminal domain to but by its carboxyl-terminal domains A. F. I. J. F. J. A. L. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). The carboxyl-terminal domains and two domains of are for its cleaving but the domain a critical in of the to and K. M. K. H. H. H. C. Y. 2003; PubMed Scopus Google Scholar, K. J. Biol. Chem. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar, P.J. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar, Y. H. A. K. 2005; PubMed Scopus Google Scholar). of K. Y. H. H. M. H. K. PubMed Scopus Google Scholar), L.A. Flannery C.R. W. Corcoran C. M. A.J. M. Morris E.A. LaVallie E.R. Biol. PubMed Scopus Google Scholar), J.M. J.M. Ross M. S. R. J. Biol. Chem. 2003; Full Text Full Text PDF PubMed Scopus Google Scholar), and M. S. C. J. Biol. Chem. 2003; Full Text Full Text PDF PubMed Scopus Google are against The carboxyl-terminal and domains of ADAMTS-4 an in its proteolytic activity M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, J. J.D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). In this we the of ADAMTS-5 carboxyl-terminal domains an similar to that used in ADAMTS-4 M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). of each ADAMTS-5 domain as as of the by to each domain to the proteolytic activities of we were to compare these activities with ADAMTS-4. Although the in 1 are of activities to the of they a of in the two The is that ADAMTS-5 was in aggrecan than ADAMTS-4 under physiological conditions. Tortorella M.D. Liu R.Q. Burn Newton R.C. Arner Biol. PubMed Scopus Google reported that ADAMTS-5 is a aggrecanase than ADAMTS-4. the other P.J. J. F. Biochem. J. 2003; PubMed Scopus Google reported that the the IGD of and by ADAMTS-5 is than that by ADAMTS-4 but that the activities of the two aggrecanases are similar on and and these reports be to in the of the enzymes used the enzymes were these influence activity as of both is observed M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, J. J.D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, C.R. W. Corcoran C. Collins-Racie L.A. S.A. LaVallie E.R. Morris E.A. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google major the ancillary domains of ADAMTS-4 and ADAMTS-5 influenced proteolytic the domain of ADAMTS-4 its activity on the aggrecanase site Glu373-Ala374 bond in the and its general proteolytic the domain of ADAMTS-5 not of its proteolytic of the domain to have a influence on ADAMTS-4 activity in but a of ADAMTS-5 activity on the The of both and domains ADAMTS-5 its activity to of the activity of In had little influence on the of of the and domains also to the of enzymes the results suggest that and domains with the of aggrecan and on the in the is by the that of the and domains to the of the of these the general aggrecanase activity of was than that of of the first and domains of ADAMTS-5 its proteolytic activity ADAMTS-4 M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), and the catalytic domain of ADAMTS-5 was essentially the catalytic domain of ADAMTS-5 the ability to cleave a and this activity was greater than that of the catalytic domain of ADAMTS-4. that are the two catalytic domains, but the for this is not at the In the ADAMTS-4 is by on the an that to the of the carboxyl-terminal domain A.J. J.D. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). In we observed a of ADAMTS-4 M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). More recently, an ADAMTS-4 that of the domain but a carboxyl-terminal been reported in J. Biol. 2006; PubMed Scopus Google Scholar). ADAMTS-4 are to We to the of ADAMTS-5 in articular cartilage with IL-1, but of ADAMTS-5 protein was not is to protein of ADAMTS-5 in the activity of ADAMTS-4 on aggrecan that we reported previously of is by 1 of at that we reported previously M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), we that of ADAMTS-5 is to degrade of the aggrecan in 1 of cartilage in a cartilage that the degradation is to Our to ADAMTS-5 at least 1 of the that at least of cartilage to be to ADAMTS-5 for a are a studies of ADAMTS-5 in the of I. M.Z. Handley C.J. Clark M. Liu C. R.A. J. Biochem. PubMed Scopus Google Scholar, Y. Clark M. R.A. Tortorella M.D. Arner E.C. J. PubMed Scopus Google and in cartilage Liu R.Q. K. S. Tortorella M.D. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Similar to ADAMTS-4 M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar, M. Pratta M. Liu R.Q. Abbaszade I. Ross H. Burn Arner J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), the on aggrecan influence the aggrecanase activity of The of the interaction of ADAMTS-4 -5 with is by to NaCl In this ADAMTS-4 was sensitive to physiological than The for the little activity of ADAMTS-5 against aggrecan at NaCl is but be to the to ADAMTS-5 and the the activity of ADAMTS-5 on aggrecan was than that of ADAMTS-4 under the physiological in aggrecanase activity ADAMTS-4 and -5 in both the activity of catalytic domains and for to of ADAMTS-4 activities on and M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), had been that aggrecanases had a for in the site M.D. Liu R.Q. Burn Newton R.C. Arner Biol. PubMed Scopus Google Scholar, M.D. Pratta M. Liu R.Q. J. Ross O.H. Abbaszade I. Burn Arner J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). a bond and carboxymethylated in Cm-Tf, but its on those are the the are we that the the site M. C. C. Itoh Y. Nagase H. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). similar was for ADAMTS-5, in this case its on was is to the catalytic domain the domain the of was similar for domain of ADAMTS-5 catalytic domain was to degrade Cm-Tf, to was We also that ADAMTS-4 and -5 are to degrade decorin, biglycan, and be that and were very degradation were the and of these activities is the of for the two aggrecanases that they have a in cartilage matrix degradation than was reported been shown to the of cartilage of aggrecan in degradation L. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus (91) Google Scholar, R. P.J. Biochem. J. 1999; PubMed Scopus Google Scholar). for this is was shown to cleave the bond of in cartilage L. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus (91) Google Scholar). studies of E.R. P.J. 2006; Full Text Full Text PDF PubMed Scopus Google that both ADAMTS-4 and -5 cleave at the and cleaved were in articular cartilage with and rheumatoid They also reported that these two aggrecanases not cleave shown in this the ability of ADAMTS-4 and to cleave is than ability to cleave Although aggrecan is a very sensitive to ADAMTS-4 and degradation of other matrix by these proteinases in as to of the articular cartilage matrix. Our in with a for ADAMTS-5 than for ADAMTS-4 in the of Although the of ADAMTS-4 and -5 in and cartilage have results J. Arthritis PubMed Scopus Google Scholar, L. C. L. S. Edwards D.R. Clark I.M. Arthritis PubMed Scopus Google Scholar), these studies that ADAMTS-5 is than ADAMTS-4. for a for ADAMTS-4 in ADAMTS-4 and in a similar to that in R. Flannery C.R. K. Yang Morris E.A. Arthritis PubMed Scopus Google Scholar), that of ADAMTS-4 not against the of ADAMTS-5 aggrecan and cartilage in two R. Flannery C.R. K. Yang Morris E.A. 2005; PubMed Scopus Google Scholar, H. C.J. Last K. P.J. A.J. 2005; PubMed Scopus Google Scholar). aggrecan degradation is also in of as with M.C. A.J. Y. A. J.D. 2006; Full Text Full Text PDF PubMed Scopus Google Scholar). Although is not the major in the of in this that ADAMTS-5 is a