title: ADAMTS1 cleaves aggrecan at multiple sites and is differentially inhibited by metalloproteinase inhibitors.
abstract: ADAMTS1 is a secreted protein that belongs to the recently described ADAMTS (a disintegrin and metalloprotease with thrombospondin repeats) family of proteases. Evaluation of ADAMTS1 catalytic activity on a panel of extracellular matrix proteins showed a restrictive substrate specificity which includes some proteoglycans. Our results demonstrated that human ADAMTS1 cleaves aggrecan at a previously shown site by its mouse homolog, but we have also identified additional cleavage sites that ultimately confirm the classification of this protease as an 'aggrecanase'. Specificity of ADAMTS1 activity was further verified when a point mutation in the zinc-binding domain abolished its catalytic effects, and latency conferred by the prodomain was also demonstrated using a furin cleavage site mutant. Suppression of ADAMTS1 activity was accomplished with a specific monoclonal antibody and some metalloprotease inhibitors, including tissue inhibitor of metalloproteinases 2 and 3. Finally, we developed an activity assay using an artificial peptide substrate based on the interglobular domain cleavage site (E(373)-A) of rat aggrecan.
authors.affiliation: Department of Molecular, Cell and Developmental Biology, Molecular Biology Institute, University of California at Los Angeles, 611, Charles E. Young Drive, Los Angeles, CA 90095, USA.
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Rodríguez-Manzaneque
authors.forename: Jennifer
authors.initials: J
authors.lastname: Westling
authors.forename: Shelley N-M
authors.initials: SN
authors.lastname: Thai
authors.forename: Alfonso
authors.initials: A
authors.lastname: Luque
authors.forename: Vera
authors.initials: V
authors.lastname: Knauper
authors.forename: Gillian
authors.initials: G
authors.lastname: Murphy
authors.forename: John D
authors.initials: JD
authors.lastname: Sandy
authors.forename: M Luisa
authors.initials: ML
authors.lastname: Iruela-Arispe
mesh_terms: D000071096:ADAMTS Proteins; D053674:Aggrecans; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D002356:Cartilage / Q000378:metabolism; D011508:Chondroitin Sulfate Proteoglycans / Q000378:metabolism; D002813:Chondrosarcoma / Q000378:metabolism; D017931:DNA Primers; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D016326:Extracellular Matrix Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D037181:Lectins, C-Type; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011509:Proteoglycans / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: C454120:ADAMTSL1 protein, human; C508320:Acan protein, mouse; C508321:Acan protein, rat; D053674:Aggrecans; D011508:Chondroitin Sulfate Proteoglycans; D017931:DNA Primers; D004791:Enzyme Inhibitors; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D011994:Recombinant Proteins; D000071096:ADAMTS Proteins; D008666:Metalloendopeptidases