title: The cysteine-rich domain regulates ADAM protease function in vivo.
abstract: ADAMs are membrane-anchored proteases that regulate cell behavior by proteolytically modifying the cell surface and ECM. Like other membrane-anchored proteases, ADAMs contain candidate "adhesive" domains downstream of their metalloprotease domains. The mechanism by which membrane-anchored cell surface proteases utilize these putative adhesive domains to regulate protease function in vivo is not well understood. We address this important question by analyzing the relative contributions of downstream extracellular domains (disintegrin, cysteine rich, and EGF-like repeat) of the ADAM13 metalloprotease during Xenopus laevis development. When expressed in embryos, ADAM13 induces hyperplasia of the cement gland, whereas ADAM10 does not. Using chimeric constructs, we find that the metalloprotease domain of ADAM10 can substitute for that of ADAM13, but that specificity for cement gland expansion requires a downstream extracellular domain of ADAM13. Analysis of finer resolution chimeras indicates an essential role for the cysteine-rich domain and a supporting role for the disintegrin domain. These and other results reveal that the cysteine-rich domain of ADAM13 cooperates intramolecularly with the ADAM13 metalloprotease domain to regulate its function in vivo. Our findings thus provide the first evidence that a downstream extracellular adhesive domain plays an active role in regulating ADAM protease function in vivo. These findings are likely relevant to other membrane-anchored cell surface proteases.
authors.affiliation: Department of Cell Biology, University of Virginia, Health Sciences Center, Charlottesville, VA 22908, USA.
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Smith
authors.forename: Alban
authors.initials: A
authors.lastname: Gaultier
authors.forename: Helene
authors.initials: H
authors.lastname: Cousin
authors.forename: Dominique
authors.initials: D
authors.lastname: Alfandari
authors.forename: Judith M
authors.initials: JM
authors.lastname: White
authors.forename: Douglas W
authors.initials: DW
authors.lastname: DeSimone
mesh_terms: D051722:ADAM Proteins; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000737:chemistry; D002678:Chimera / Q000235:genetics; D003545:Cysteine / Q000737:chemistry*; D019483:Disintegrins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005088:Exocrine Glands / Q000196:embryology / Q000473:pathology; D005109:Extracellular Matrix / Q000378:metabolism; D000072196:Fertilins; D018507:Gene Expression Regulation, Developmental; D015971:Gene Expression Regulation, Enzymologic; D006965:Hyperplasia; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008954:Models, Biological; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009432:Neural Crest / Q000166:cytology / Q000378:metabolism; D017434:Protein Structure, Tertiary; D012333:RNA, Messenger / Q000378:metabolism; D016415:Sequence Alignment; D029867:Xenopus Proteins; D014982:Xenopus laevis / Q000196:embryology / Q000235:genetics / Q000378:metabolism
chemical_list: D019483:Disintegrins; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D012333:RNA, Messenger; D029867:Xenopus Proteins; D051722:ADAM Proteins; C104937:ADAM33 protein, Xenopus; D000072196:Fertilins; D008666:Metalloendopeptidases; D003545:Cysteine