title: Neutrophil collagenase (MMP-8) cleaves at the aggrecanase site E373-A374 in the interglobular domain of cartilage aggrecan.
abstract: Native and recombinant neutrophil collagenase (MMP-8) was shown to cleave at the E373-A374 'aggrecanase' site in the interglobular domain of aggrecan. The time course of digestion in vitro showed that MMP-8 cleaved initially at N341-F342, the predominant metalloproteinase site, before cleaving at the E373-A374 site. A synthetic peptide, IPENFFG, inhibited cleavage at E373-A374 but not N341-F342 in vitro, indicating that the E373-A374 sequence was a less preferred site for MMP-8 cleavage than N341-F342. IPENFFG also inhibited release of A374 RGSVI fragments from cartilage in explant culture, suggesting that a metalloproteinase cleaved at the aggrecanase site in situ. The possibility remains that 'aggrecanase' may be a metalloproteinase in cartilage.
authors.affiliation: University of Melbourne, Department of Medicine, Royal Melbourne Hospital, Parkville, Australia.
authors.forename: A J
authors.initials: AJ
authors.lastname: Fosang
authors.forename: K
authors.initials: K
authors.lastname: Last
authors.forename: P J
authors.initials: PJ
authors.lastname: Neame
authors.forename: G
authors.initials: G
authors.lastname: Murphy
authors.forename: V
authors.initials: V
authors.lastname: Knäuper
authors.forename: H
authors.initials: H
authors.lastname: Tschesche
authors.forename: C E
authors.initials: CE
authors.lastname: Hughes
authors.forename: B
authors.initials: B
authors.lastname: Caterson
authors.forename: T E
authors.initials: TE
authors.lastname: Hardingham
mesh_terms: D053674:Aggrecans; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D002356:Cartilage / Q000737:chemistry*; D017364:Collagenases / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism*; D016326:Extracellular Matrix Proteins; D006801:Humans; D037181:Lectins, C-Type; D020784:Matrix Metalloproteinase 8; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000201:enzymology; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D011509:Proteoglycans / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013379:Substrate Specificity; D013552:Swine
chemical_list: D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D010446:Peptide Fragments; D011509:Proteoglycans; D011994:Recombinant Proteins; D010450:Endopeptidases; D017364:Collagenases; D020784:Matrix Metalloproteinase 8; C083641:aggrecanase