title: Interactions of a laminin-binding peptide from a 33-kDa protein related to the 67-kDa laminin receptor with laminin and melanoma cells are heparin-dependent.
abstract: A laminin-binding peptide (peptide G), predicted from the cDNA sequence for a 33-kDa protein related to the 67-kDa laminin receptor, specifically inhibits binding of laminin to heparin and sulfatide. Since the peptide binds directly to heparin and inhibits interaction of another heparin-binding protein with the same sulfated ligands, this inhibition is due to direct competition for binding to sulfated glycoconjugates rather than an indirect effect of interaction with the binding site on laminin for the 67-kDa receptor. Direct binding of laminin to the peptide is also inhibited by heparin. This interaction may result from contamination of the laminin with heparan sulfate, as binding is enhanced by the addition of substoichiometric amounts of heparin but inhibited by excess heparin and two heparin-binding proteins. Furthermore, laminin binds more avidly to a heparin-binding peptide derived from thrombospondin than to the putative receptor peptide. Adhesion of A2058 melanoma cells on immobilized peptide G is also heparin-dependent, whereas adhesion of the cells on laminin is not. Antibodies to the beta 1-integrin chain or laminin block adhesion of the melanoma cells to laminin but not to peptide G. Thus, the reported inhibition of melanoma cell adhesion to endothelial cells by peptide G may result from inhibition of binding of laminin or other proteins to sulfated glycoconjugate receptors rather than from specific inhibition of laminin binding to the 67-kDa receptor.
authors.affiliation: Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892.
authors.forename: N H
authors.initials: NH
authors.lastname: Guo
authors.forename: H C
authors.initials: HC
authors.lastname: Krutzsch
authors.forename: T
authors.initials: T
authors.lastname: Vogel
authors.forename: D D
authors.initials: DD
authors.lastname: Roberts
mesh_terms: D000595:Amino Acid Sequence; D001667:Binding, Competitive; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D006493:Heparin / Q000378:metabolism* / Q000494:pharmacology; D006801:Humans; D007797:Laminin / Q000378:metabolism*; D008545:Melanoma / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011498:Protein Precursors; D011971:Receptors, Immunologic / Q000378:metabolism*; D017474:Receptors, Laminin; D013433:Sulfoglycosphingolipids / Q000494:pharmacology; D019699:Thrombospondins; D014407:Tumor Cells, Cultured
chemical_list: D007797:Laminin; D010455:Peptides; D010980:Platelet Membrane Glycoproteins; D011498:Protein Precursors; D011971:Receptors, Immunologic; D017474:Receptors, Laminin; D013433:Sulfoglycosphingolipids; D019699:Thrombospondins; D006493:Heparin