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Interaction of Fibronectin With Cultured Human Endothelial Cells: Characterization of the Specific Receptor

In this study we provide a characterization of the fibronectin (FN) binding to endothelial cells (EC), and we identify the FN binding site on these cells. 125I-FN binding to EC in suspension was time dependent and reached a plateau at 4 h. Cold FN inhibited this interaction in a concentration-depend...

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Published in:Blood 1989-05, Vol.73 (6), p.1576-1585
Main Authors: Conforti, Grazia, Zanetti, Adriana, Colella, Silvia, Abbadini, Marzia, Marchisio, Pier Carlo, Pytela, Robert, Giancotti, Filippo, Tarone, Guido, Languino, Lucia Raffaella, Dejana, Elisabetta
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container_end_page 1585
container_issue 6
container_start_page 1576
container_title Blood
container_volume 73
creator Conforti, Grazia
Zanetti, Adriana
Colella, Silvia
Abbadini, Marzia
Marchisio, Pier Carlo
Pytela, Robert
Giancotti, Filippo
Tarone, Guido
Languino, Lucia Raffaella
Dejana, Elisabetta
description In this study we provide a characterization of the fibronectin (FN) binding to endothelial cells (EC), and we identify the FN binding site on these cells. 125I-FN binding to EC in suspension was time dependent and reached a plateau at 4 h. Cold FN inhibited this interaction in a concentration-dependent way, but vitronectin, fibrinogen, and IgG were poorly effective. About 80% of the total FN associated to EC at the equilibrium was specifically bound; of this, 60% was reversibly bound, while 20% appeared to be internalized. The FN binding was saturable and an apparent dissociation constant of about 0.23 � 10-6 mol/L and a maximal number of binding sites of about 9.8 � 105 was estimated from binding isotherms. Autoradiography data showed that EC-associated 125I-FN was all in high mol wt form that did not enter the gel. We then characterize the FN receptor (FNR) in EC. An antiserum to the FNR isolated from human placenta inhibited FN binding to EC by 89%, and using the immunoblotting technique, it recognized two bands in the EC detergent extract of mol wt 125/160 Kd. This antiserum also recognized the EC membrane protein complex eluted from the FN affinity column by an arg-gly-asp (RGD) peptide. When this complex was included into liposomes, it poorly bound to FN. However, the binding was strikingly increased by addition of Mn in the buffer and was specific for FN in respect to other substrata. These data define the FN binding site in EC and indicate that it is functionally and structurally related to that isolated from human placenta.
doi_str_mv 10.1182/blood.V73.6.1576.1576
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identifier ISSN: 0006-4971
ispartof Blood, 1989-05, Vol.73 (6), p.1576-1585
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1528-0020
language eng
recordid cdi_proquest_miscellaneous_78970491
source ScienceDirect®
subjects Analytical, structural and metabolic biochemistry
Antigens, Surface
Biological and medical sciences
Cell Adhesion Molecules
Cells, Cultured
Endothelium, Vascular - physiology
Extracellular Matrix - metabolism
Fibrinogen - metabolism
Fibronectins - physiology
Fundamental and applied biological sciences. Psychology
Glycoproteins
Glycoproteins - metabolism
Humans
In Vitro Techniques
Kinetics
Liposomes
Molecular Weight
Proteins
Receptors, Fibronectin
Receptors, Immunologic - physiology
Vitronectin
title Interaction of Fibronectin With Cultured Human Endothelial Cells: Characterization of the Specific Receptor
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