title: Characterization of human aggrecanase 2 (ADAM-TS5): substrate specificity studies and comparison with aggrecanase 1 (ADAM-TS4).
abstract: ADAM-TS5 (aggrecanase 2), one of two cartilage aggrecanases is a member of the ADAM protein family. Like ADAM-TS4 (aggrecanase 1) the enzyme cleaves cartilage aggrecan at the Glu(373)-Ala(374) bond, a marker of aggrecanase activity. In this study we have characterized the substrate specificity of ADAM-TS5 and compared it with that of ADAM-TS4. The recombinant human ADAM-TS5, like ADAM-TS4 cleaves aggrecan at Glu(1480)-Gly(1481), Glu(1667)-Gly(1668), Glu(1771)-Ala(1772) and Glu(1871)-Leu(1872) bonds more readily than at the Glu(373)-Ala(374) bond. In addition, ADAM-TS5 exhibited an additional site of cleavage in the region spanning residues Gly(1481) and Glu(1667), representing a unique cleavage of ADAM-TS5. ADAM-TS5 cleaved aggrecan approximately 2-fold slower than ADAM-TS4. Neither ADAM-TS5 nor ADAM-TS4 was able to cleave the extracellular matrix proteins fibronectin, thrombospondin, type I collagen, type II collagen, gelatin or general protein substrates such as casein and transferrin. Finally, the zymogen of stromelysin (MMP-3) was not activated by either ADAM-TS4 or ADAM-TS5.
authors.affiliation: Department of Inflammatory Diseases Research, DuPont Pharmaceuticals Company, Wilmington, DE 19880-0400, USA. micky.d.tortorella@pharmacia.com
authors.forename: Micky D
authors.initials: MD
authors.lastname: Tortorella
authors.forename: Rui-Qin
authors.initials: RQ
authors.lastname: Liu
authors.forename: Timothy
authors.initials: T
authors.lastname: Burn
authors.forename: Robert C
authors.initials: RC
authors.lastname: Newton
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Arner
mesh_terms: D051722:ADAM Proteins; D000071121:ADAMTS4 Protein; D000071116:ADAMTS5 Protein; D053674:Aggrecans; D000818:Animals; D002460:Cell Line; D004330:Drosophila / Q000166:cytology; D016326:Extracellular Matrix Proteins / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D037181:Lectins, C-Type; D008666:Metalloendopeptidases / Q000378:metabolism* / Q000494:pharmacology; D011348:Procollagen N-Endopeptidase; D011509:Proteoglycans / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism / Q000494:pharmacology; D013379:Substrate Specificity; D013997:Time Factors
chemical_list: D053674:Aggrecans; D016326:Extracellular Matrix Proteins; D037181:Lectins, C-Type; D011509:Proteoglycans; D011994:Recombinant Proteins; D051722:ADAM Proteins; D000071116:ADAMTS5 Protein; C121532:ADAMTS5 protein, human; D008666:Metalloendopeptidases; D011348:Procollagen N-Endopeptidase; D000071121:ADAMTS4 Protein