title: Characterization of ADAMTS-9 and ADAMTS-20 as a distinct ADAMTS subfamily related to Caenorhabditis elegans GON-1.
abstract: We demonstrate that in humans, two metalloproteases, ADAMTS-9 (1935 amino acids) and ADAMTS-20 (1911 amino acids) are orthologs of GON-1, an ADAMTS protease required for gonadal morphogenesis in Caenorhabditis elegans. ADAMTS-9 and ADAMTS-20 have an identical modular structure, are distinct in possessing 15 TSRs and a unique C-terminal domain, and have a similar gene structure, suggesting that they comprise a new subfamily of human ADAMTS proteases. ADAMTS20 is very sparingly expressed, although it is detectable in epithelial cells of the breast and lung. However, ADAMTS9 is highly expressed in embryonic and adult tissues, and therefore we characterized the ADAMTS-9 protein further. Although the ADAMTS-9 zymogen has many proprotein convertase processing sites, pulse-chase analysis, site-directed mutagenesis, and amino acid sequencing demonstrated that maturation to the active form occurs by selective proprotein convertase (e.g. furin) cleavage of the Arg(287)-Phe(288) bond. Although lacking a transmembrane sequence, ADAMTS-9 is retained near the cell surface as well as in the ECM of transiently transfected COS-1 and 293 cells. COS-1 cells transfected with ADAMTS9 (but not vector-transfected cells) proteolytically cleaved bovine versican and aggrecan core proteins at the Glu(441)-Ala(442) bond of versican V1 and the Glu(1771)-Ala(1772) bond of aggrecan, respectively. In contrast, the ADAMTS-9 catalytic domain alone was neither localized to the cell surface nor able to confer these proteolytic activities on cells, demonstrating that the ancillary domains of ADAMTS-9, including the TSRs, are required both for specific extracellular localization and for its versicanase and aggrecanase activities.
authors.affiliation: Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic Foundation, Cleveland, Ohio 44195, USA.
authors.forename: Robert P T
authors.initials: RP
authors.lastname: Somerville
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Longpre
authors.forename: Katherine A
authors.initials: KA
authors.lastname: Jungers
authors.forename: J Michael
authors.initials: JM
authors.lastname: Engle
authors.forename: Monique
authors.initials: M
authors.lastname: Ross
authors.forename: Stephen
authors.initials: S
authors.lastname: Evanko
authors.forename: Thomas N
authors.initials: TN
authors.lastname: Wight
authors.forename: Richard
authors.initials: R
authors.lastname: Leduc
authors.forename: Suneel S
authors.initials: SS
authors.lastname: Apte
mesh_terms: D051722:ADAM Proteins; D000071096:ADAMTS Proteins; D000071119:ADAMTS9 Protein; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001120:Arginine / Q000737:chemistry; D015152:Blotting, Northern; D015153:Blotting, Western; D019556:COS Cells; D017173:Caenorhabditis elegans; D029742:Caenorhabditis elegans Proteins; D020134:Catalytic Domain; D002417:Cattle; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D010450:Endopeptidases / Q000096:biosynthesis / Q000737:chemistry / Q000145:classification*; D005973:Glutamine / Q000737:chemistry; D006801:Humans; D017403:In Situ Hybridization; D008666:Metalloendopeptidases / Q000096:biosynthesis / Q000737:chemistry* / Q000145:classification*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D010649:Phenylalanine / Q000737:chemistry; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013997:Time Factors; D014018:Tissue Distribution; D014162:Transfection
chemical_list: D029742:Caenorhabditis elegans Proteins; D018076:DNA, Complementary; D005973:Glutamine; D010649:Phenylalanine; D001120:Arginine; D010450:Endopeptidases; D051722:ADAM Proteins; D000071096:ADAMTS Proteins; C473773:ADAMTS20 protein, human; D000071119:ADAMTS9 Protein; C412869:ADAMTS9 protein, human; C119674:GON-1 protein, C elegans; D008666:Metalloendopeptidases; C083641:aggrecanase