title: Cloning and characterization of ADAMTS11, an aggrecanase from the ADAMTS family.
abstract: 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 inflammation-associated 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.
authors.affiliation: Department of Applied Biotechnology, The DuPont Pharmaceuticals Company, Experimental Station, Wilmington, Delaware 19880, USA.
authors.forename: I
authors.initials: I
authors.lastname: Abbaszade
authors.forename: R Q
authors.initials: RQ
authors.lastname: Liu
authors.forename: F
authors.initials: F
authors.lastname: Yang
authors.forename: S A
authors.initials: SA
authors.lastname: Rosenfeld
authors.forename: O H
authors.initials: OH
authors.lastname: Ross
authors.forename: J R
authors.initials: JR
authors.lastname: Link
authors.forename: D M
authors.initials: DM
authors.lastname: Ellis
authors.forename: M D
authors.initials: MD
authors.lastname: Tortorella
authors.forename: M A
authors.initials: MA
authors.lastname: Pratta
authors.forename: J M
authors.initials: JM
authors.lastname: Hollis
authors.forename: R
authors.initials: R
authors.lastname: Wynn
authors.forename: J L
authors.initials: JL
authors.lastname: Duke
authors.forename: H J
authors.initials: HJ
authors.lastname: George
authors.forename: M C
authors.initials: MC
authors.lastname: Hillman
authors.forename: K
authors.initials: K
authors.lastname: Murphy
authors.forename: B H
authors.initials: BH
authors.lastname: Wiswall
authors.forename: R A
authors.initials: RA
authors.lastname: Copeland
authors.forename: C P
authors.initials: CP
authors.lastname: Decicco
authors.forename: R
authors.initials: R
authors.lastname: Bruckner
authors.forename: H
authors.initials: H
authors.lastname: Nagase
authors.forename: Y
authors.initials: Y
authors.lastname: Itoh
authors.forename: R C
authors.initials: RC
authors.lastname: Newton
authors.forename: R L
authors.initials: RL
authors.lastname: Magolda
authors.forename: J M
authors.initials: JM
authors.lastname: Trzaskos
authors.forename: T C
authors.initials: TC
authors.lastname: Burn
mesh_terms: D051722:ADAM Proteins; D000071116:ADAMTS5 Protein; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003001:Cloning, Molecular; D018076:DNA, Complementary; D010450:Endopeptidases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D010450:Endopeptidases; D051722:ADAM Proteins; D000071116:ADAMTS5 Protein; C121532:ADAMTS5 protein, human; D008666:Metalloendopeptidases; C083641:aggrecanase