Human melanoma cells express a novel integrin receptor for laminin.
This study sought to determine whether human melanoma cells express integrin-related receptors that mediate their adhesion to laminin. We found that antibodies against the integrin beta 1 chain blocked cell attachment to laminin-coated surfaces. Furthermore, immunofluorescence staining demonstrated beta 1 complexes in vinculin-positive focal adhesion plaques on the basal surface of cells attached to laminin substrates. Chromatography of detergent extracts of 125I-surface-labeled cells on laminin-Sepharose columns recovered two major laminin-binding proteins (100 and 130 kDa, reduced) that bound with high affinity to the columns and were eluted with EDTA. Both proteins were specifically immunoprecipitated from column fractions with monoclonal and polyclonal antibodies to the integrin beta 1 subunit, indicating that they form a noncovalent heterodimer complex. The alpha-like subunit is composed of a 30-kDa light chain that is joined by a disulfide bond to the 100-kDa heavy chain. This complex was not recovered from columns of fibronectin- or collagen type I- or IV-Sepharose. Laminin-binding by the alpha beta 1 complex was independent of Arg-Gly-Asp or Tyr-Ile-Gly-Ser-Arg-like sequences, but required the presence of divalent cations. The 100-kDa alpha-like subunit was electrophoretically and immunochemically distinct from the other known alpha subunits, alpha 1-alpha 6. The results indicate that human melanoma cells express a novel laminin-specific integrin beta 1 complex which may mediate the cells' interactions with this ligand.
Kramer
R H
RH
Department of Stomatology, University of California, San Francisco 94143.
McDonald
K A
KA
Vu
M P
MP
D000911:Antibodies, Monoclonal; D002448:Cell Adhesion; D002460:Cell Line; D002846:Chromatography, Affinity; D005455:Fluorescent Antibody Technique; D006801:Humans; D016023:Integrins; D007700:Kinetics; D007797:Laminin; D046911:Macromolecular Substances; D008545:Melanoma / Q000503:physiopathology*; D008562:Membrane Glycoproteins / Q000032:analysis / Q000276:immunology; D008970:Molecular Weight; D011971:Receptors, Immunologic / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D017474:Receptors, Laminin
D000911:Antibodies, Monoclonal; D016023:Integrins; D007797:Laminin; D046911:Macromolecular Substances; D008562:Membrane Glycoproteins; D011971:Receptors, Immunologic; D017474:Receptors, Laminin