title: Localization of the ganglioside-binding site of fibronectin.
abstract: It has been demonstrated via biological assays that fibronectin possesses a receptor for gangliosides that is involved in cell adhesion and restoration of the normal morphology of transformed cells. In this study, fluorescence polarization has been employed to monitor the binding of ganglioside oligosaccharide to fibronectin. Parameters involved in ganglioside oligosaccharide binding to fibronectin are described and compared to the interaction of heparin with fibronectin. A Kd of 1.4 X 10(-8) mol/liter has been calculated, and it is demonstrated that labeled ganglioside oligosaccharides can be eluted from fibronectin with either unlabeled ganglioside oligosaccharides or 4 M urea. Using the fluorescence polarization assay developed in this study for measurement of ganglioside binding to fibronectin, it is demonstrated that gangliosides bind to the 31,000-dalton amino terminal heparin-binding domain of fibronectin. A ganglioside-Sepharose affinity column has been constructed which specifically binds the 31,000-dalton amino terminal fragment of fibronectin. The localization of the ganglioside receptor to the amino terminal domain of fibronectin indicates that the ganglioside receptor is distinct from the putative fibronectin cell surface receptor which is located near the center of the fibronectin molecule.
mesh_terms: D001665:Binding Sites; D005353:Fibronectins / Q000378:metabolism*; D005452:Fluoresceins; D005454:Fluorescence Polarization; D005456:Fluorescent Dyes; D005732:Gangliosides / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009844:Oligosaccharides / Q000378:metabolism; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000378:metabolism; D013696:Temperature
chemical_list: D005353:Fibronectins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005732:Gangliosides; D009844:Oligosaccharides; D010446:Peptide Fragments; C027101:fluorescein thiocarbamylethylenediamine; D006493:Heparin