Transmembrane collagen XVII, an epithelial adhesion protein, is shed from the cell surface by ADAMs.
Collagen XVII, a type II transmembrane protein and epithelial adhesion molecule, can be proteolytically shed from the cell surface to generate a soluble collagen. Here we investigated the release of the ectodomain and identified the enzymes involved. After surface biotinylation of keratinocytes, the ectodomain was detectable in the medium within minutes and remained stable for >48 h. Shedding was enhanced by phorbol esters and inhibited by metalloprotease inhibitors, including hydroxamates and TIMP-3, but not by inhibitors of other protease classes or by TIMP-2. This profile implicated MMPs or ADAMs as candidate sheddases. MMP-2, MMP-9 and MT1-MMP were excluded, but TACE, ADAM-10 and ADAM-9 were shown to be expressed in keratinocytes and to be actively involved. Transfection with cDNAs for the three ADAMs resulted in increased shedding and, vice versa, in TACE-deficient cells shedding was significantly reduced, indicating that transmembrane collagen XVII represents a novel class of substrates for ADAMs. Functionally, release of the ectodomain of collagen XVII from the cell surface was associated with altered keratinocyte motility in vitro.
D000918:Antibody Specificity; D001324:Autoantigens / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D001483:Base Sequence; D019921:Biotinylation; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology*; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry / Q000276:immunology / Q000502:physiology*; D003598:Cytoskeletal Proteins; D017931:DNA Primers; D000071821:Dystonin; D010450:Endopeptidases / Q000378:metabolism; D006801:Humans; D015603:Keratinocytes / Q000502:physiology*; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D011480:Protease Inhibitors / Q000494:pharmacology; D011994:Recombinant Proteins / Q000378:metabolism; D019716:Tissue Inhibitor of Metalloproteinase-2 / Q000494:pharmacology; D019717:Tissue Inhibitor of Metalloproteinase-3 / Q000494:pharmacology; D014162:Transfection; D000097805:Collagen Type XVII
D001324:Autoantigens; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D003094:Collagen; D003598:Cytoskeletal Proteins; D017931:DNA Primers; D000071821:Dystonin; D010450:Endopeptidases; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases; D009419:Nerve Tissue Proteins; D024041:Non-Fibrillar Collagens; D011480:Protease Inhibitors; D011994:Recombinant Proteins; D019716:Tissue Inhibitor of Metalloproteinase-2; D019717:Tissue Inhibitor of Metalloproteinase-3; D000097805:Collagen Type XVII; C486268:DST protein, human