title: The cysteine-rich domain of human ADAM 12 supports cell adhesion through syndecans and triggers signaling events that lead to beta1 integrin-dependent cell spreading.
abstract: The ADAMs (a disintegrin and metalloprotease) family of proteins is involved in a variety of cellular interactions, including cell adhesion and ecto- domain shedding. Here we show that ADAM 12 binds to cell surface syndecans. Three forms of recombinant ADAM 12 were used in these experiments: the cys-teine-rich domain made in Escherichia coli (rADAM 12-cys), the disintegrin-like and cysteine-rich domain made in insect cells (rADAM 12-DC), and full-length human ADAM 12-S tagged with green fluorescent protein made in mammalian cells (rADAM 12-GFP). Mesenchymal cells specifically and in a dose-dependent manner attach to ADAM 12 via members of the syndecan family. After binding to syndecans, mesenchymal cells spread and form focal adhesions and actin stress fibers. Integrin beta1 was responsible for cell spreading because function-blocking monoclonal antibodies completely inhibited cell spreading, and chondroblasts lacking beta1 integrin attached but did not spread. These data suggest that mesenchymal cells use syndecans as the initial receptor for the ADAM 12 cysteine-rich domain-mediated cell adhesion, and then the beta1 integrin to induce cell spreading. Interestingly, carcinoma cells attached but did not spread on ADAM 12. However, spreading could be efficiently induced by the addition of either 1 mM Mn(2+) or the beta1 integrin-activating monoclonal antibody 12G10, suggesting that in these carcinoma cells, the ADAM 12-syndecan complex fails to modulate the function of beta1 integrin.
authors.affiliation: The Institute of Molecular Pathology, University of Copenhagen, 2100 Copenhagen, Denmark.
authors.forename: K
authors.initials: K
authors.lastname: Iba
authors.forename: R
authors.initials: R
authors.lastname: Albrechtsen
authors.forename: B
authors.initials: B
authors.lastname: Gilpin
authors.forename: C
authors.initials: C
authors.lastname: Fröhlich
authors.forename: F
authors.initials: F
authors.lastname: Loechel
authors.forename: A
authors.initials: A
authors.lastname: Zolkiewska
authors.forename: K
authors.initials: K
authors.lastname: Ishiguro
authors.forename: T
authors.initials: T
authors.lastname: Kojima
authors.forename: W
authors.initials: W
authors.lastname: Liu
authors.forename: J K
authors.initials: JK
authors.lastname: Langford
authors.forename: R D
authors.initials: RD
authors.lastname: Sanderson
authors.forename: C
authors.initials: C
authors.lastname: Brakebusch
authors.forename: R
authors.initials: R
authors.lastname: Fässler
authors.forename: U M
authors.initials: UM
authors.lastname: Wewer
mesh_terms: D051722:ADAM Proteins; D000072199:ADAM12 Protein; D000199:Actins / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001943:Breast Neoplasms; D002448:Cell Adhesion / Q000502:physiology; D048429:Cell Size / Q000187:drug effects / Q000502:physiology; D002642:Chick Embryo; D019902:Chondrocytes / Q000166:cytology / Q000378:metabolism; D003110:Colonic Neoplasms; D003545:Cysteine; D003599:Cytoskeleton / Q000502:physiology; D006801:Humans; D019012:Integrin beta1 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology; D008666:Metalloendopeptidases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008815:Mice, Inbred Strains; D018482:Muscle, Skeletal / Q000166:cytology; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D012516:Osteosarcoma; D017434:Protein Structure, Tertiary; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism*; D020239:Receptor Cross-Talk / Q000502:physiology; D012208:Rhabdomyosarcoma; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D013314:Stress, Mechanical; D053667:Syndecans; D014407:Tumor Cells, Cultured / Q000166:cytology / Q000378:metabolism
chemical_list: D000199:Actins; D000911:Antibodies, Monoclonal; D019012:Integrin beta1; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D011509:Proteoglycans; D053667:Syndecans; D051722:ADAM Proteins; D000072199:ADAM12 Protein; C000606480:ADAM12 protein, human; D008666:Metalloendopeptidases; D008274:Magnesium; D003545:Cysteine