Biosynthetic and functional properties of an Arg-Gly-Asp-directed receptor involved in human melanoma cell attachment to vitronectin, fibrinogen, and von Willebrand factor.
M21 human melanoma cells express an Arg-Gly-Asp-directed adhesion receptor composed of noncovalently associated alpha and beta chains. To establish the structural and functional properties of this receptor on M21 human melanoma cells, stable variant cell lines were selected that express altered alpha chain levels. One of these variants, M21-L, fails to synthesize alpha chain protein or its mRNA, yet does produce normal levels of the beta chain. In these cells the beta chain does not reach the cell surface but rather accumulates within the cell. M21-L cells lacking the alpha chain are incapable of attaching to vitronectin, von Willebrand factor, fibrinogen, or an Arg-Gly-Asp-containing heptapeptide yet attach normally to fibronectin, whereas the unselected M21 cells attach to all of these adhesive proteins. In addition, a monoclonal antibody, LM609 generated to a functional site on the intact receptor, is capable of preventing M21 cell attachment to vitronectin, von Willebrand factor, fibrinogen, and the Arg-Gly-Asp peptide but not to fibronectin. Following a 2-min biosynthetic pulse-label, the newly synthesized alpha chain remains in free form for 5 min and then associates with previously synthesized beta chain present in an intracellular pool. Once oligomerization takes place, the receptor gains the capacity to recognize Arg-Gly-Asp, and at this time the epitope recognized by monoclonal antibody LM609 is formed.
D002448:Cell Adhesion; D002460:Cell Line; D002846:Chromatography, Affinity; D005340:Fibrinogen / Q000378:metabolism*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D008545:Melanoma / Q000276:immunology*; D009842:Oligopeptides / Q000378:metabolism*; D011971:Receptors, Immunologic / Q000096:biosynthesis*; D018000:Receptors, Peptide; D014407:Tumor Cells, Cultured / Q000378:metabolism; D019096:Vitronectin; D014841:von Willebrand Factor / Q000378:metabolism*
D006023:Glycoproteins; D009842:Oligopeptides; D011971:Receptors, Immunologic; D018000:Receptors, Peptide; D019096:Vitronectin; C051382:arginyl-glycyl-aspartic acid directed cell adhesion receptor; D014841:von Willebrand Factor; C047981:arginyl-glycyl-aspartic acid; D005340:Fibrinogen