Fibronectin and vitronectin regulate the organization of their respective Arg-Gly-Asp adhesion receptors in cultured human endothelial cells.
Human umbilical vein endothelial cells (ECs) adhere in vitro to proteins of the extracellular matrix including fibronectin (fn) and vitronectin (vn). Specific receptors for fn and vn have been previously characterized. These receptors belong to a family of membrane glycoproteins characterized (a) by being a transmembrane complex of two noncovalently linked subunits and (b) by recognizing the tripeptide Arg-Gly-Asp on their respective ligands. In this paper we investigated how vn and fn control the organization of their respective receptors over the surface of ECs. It was found that the clustering of individual receptors and the organization thereafter of focal contacts occurred only when ECs were exposed to the specific ligand and did not occur on the opposite ligand. The shape of receptor clusters was slightly different and a colocalization of the two receptors was found when ECs were cultured on a mixed matrix of fn plus vn. Adhesion was selectively inhibited by vn or fn receptor antibodies on their respective substrates. The clustering of both receptors preceded the association of vinculin with focal contacts and stress fiber formation. Also, the vn receptor, in the absence of associated fn receptor, was capable of inducing the organization of the membrane-microfilament interaction complex. Overall, these results indicate that individual matrix ligands induce only the clustering of their respective membrane receptors. The clustering of only one receptor is capable of supporting the subsequent formation of focal contacts and the local assembly of related cytoskeletal proteins.
Dejana
E
E
Istituto di Richerche Farmacologiche Mario Negri, Milano, Italy.
Colella
S
S
Conforti
G
G
Abbadini
M
M
Gaboli
M
M
Marchisio
P C
PC
D001798:Blood Proteins / Q000502:physiology*; D002448:Cell Adhesion; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000378:metabolism*; D005353:Fibronectins / Q000502:physiology*; D005455:Fluorescent Antibody Technique; D006023:Glycoproteins / Q000502:physiology*; D006801:Humans; D007118:Immunoassay; D008024:Ligands; D009124:Muscle Proteins / Q000032:analysis / Q000378:metabolism; D011233:Precipitin Tests; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic / Q000032:analysis / Q000378:metabolism*; D018000:Receptors, Peptide; D019030:Receptors, Vitronectin; D016596:Vinculin; D019096:Vitronectin
D001798:Blood Proteins; D005353:Fibronectins; D006023:Glycoproteins; D008024:Ligands; D009124:Muscle Proteins; D017469:Receptors, Fibronectin; D011971:Receptors, Immunologic; D018000:Receptors, Peptide; D019030:Receptors, Vitronectin; D019096:Vitronectin; C051382:arginyl-glycyl-aspartic acid directed cell adhesion receptor; D016596:Vinculin