Lack of fibronectin-binding plasma membrane proteins may explain defective pericellular matrix formation in transformed fibroblasts and fibrosarcoma cells.
Affinity of iodinated fibronectin (Fn) and its defined proteolytic fragments to electrophoretically separated polypeptides of normal and malignant cells was studied in an overlay assay. Cellular 125I-Fn and a major 125I-Fn fragment (Mr 120,000-140,000), containing the cell-binding site, revealed in fibroblasts Mr 170,000, Mr 140,000, and Mr 47,000 Fn-binding polypeptides of which the first two could also be found in the plasma membrane preparations. Binding of 125I-Fns to Mr 170,000 and Mr 140,000 polypeptides was inhibited by the synthetic peptide Arg-Gly-Asp-Ser and to all 3 polypeptides by Fns and Mr 120,000-140,000 fibronectin fragment. Both fibrosarcoma cells and SV40-virus-transformed fibroblasts appeared to lack the Mr 140,000 Fn-binding polypeptide. Binding was similar when Fn from normal fibroblasts or fibrosarcoma cells was used in the assay, while plasma 125I-Fn had weaker affinity towards the Mr 140,000 polypeptide. Instead, proteolytic Fn-fragments, lacking the cell binding site, did not bind to any proteins in the assay. Radioactive cell-surface labelling showed differences in the corresponding surface polypeptide profiles of normal and malignant cells. The results suggest that the failure of pericellular matrix deposition in malignant cells could be due to either defective surface exposition or defective binding property of the Fn-receptor-like polypeptides.
D001345:Autoradiography; D001665:Binding Sites; D002471:Cell Transformation, Neoplastic; D004591:Electrophoresis, Polyacrylamide Gel; D005109:Extracellular Matrix / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism*; D005354:Fibrosarcoma / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008970:Molecular Weight; D009842:Oligopeptides / Q000494:pharmacology; D011485:Protein Binding
D005353:Fibronectins; D008565:Membrane Proteins; D009842:Oligopeptides; C046617:arginyl-glycyl-aspartyl-serine