title: Conformational regulation of the fibronectin binding and alpha 3beta 1 integrin-mediated adhesive activities of thrombospondin-1.
abstract: The recognition of extracellular matrix components can be regulated by conformational changes that alter the activity of cell surface integrins. We now demonstrate that conformational regulation of the matrix glycoprotein thrombospondin-1 (TSP1) can also modulate its binding to an integrin receptor. F18 1G8 is a conformation-sensitive TSP1 antibody that binds weakly to soluble TSP1 in the presence of divalent cations. However, binding of the antibody to melanoma cells was strongly stimulated by adding exogenous TSP1 in the presence of calcium, suggesting that TSP1 undergoes a conformational change following its binding to the cell surface. This conformation was not induced by known cell surface TSP1 receptors, whereas binding of F18 was stimulated when TSP1 bound to fibronectin but not to heparin or fibrinogen. Conversely, binding of F18 to TSP1 enhanced TSP1 binding to fibronectin. Exogenous fibronectin also stimulated TSP1-dependent binding of F18 to melanoma cells. Binding of the fibronectin-TSP1 complex to melanoma cells was mediated by alpha4beta1 and alpha5beta1 integrins. Furthermore, binding to F18 or fibronectin strongly enhanced the adhesive activity of immobilized TSP1 for some cell types. This enhancement of adhesion was mediated by alpha3beta1 integrin and required that the alpha3beta1 integrin be in an active state. Fibronectin also enhanced TSP1 binding to purified alpha3beta1 integrin. Therefore, both fibronectin and the F18 antibody induce conformational changes in TSP1 that enhance the ability of TSP1 to be recognized by alpha3beta1 integrin. The conformational and functional regulation of TSP1 activity by fibronectin represents a novel mechanism for extracellular signal transduction.
authors.affiliation: Laboratory of Pathology, NCI, National Institutes of Health and Hematology Service, Clinical Center, National Institutes of Health, Bethesda, Maryland 20892, USA.
authors.forename: R G
authors.initials: RG
authors.lastname: Rodrigues
authors.forename: N
authors.initials: N
authors.lastname: Guo
authors.forename: L
authors.initials: L
authors.lastname: Zhou
authors.forename: J M
authors.initials: JM
authors.lastname: Sipes
authors.forename: S B
authors.initials: SB
authors.lastname: Williams
authors.forename: N S
authors.initials: NS
authors.lastname: Templeton
authors.forename: H R
authors.initials: HR
authors.lastname: Gralnick
authors.forename: D D
authors.initials: DD
authors.lastname: Roberts
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002448:Cell Adhesion; D004305:Dose-Response Relationship, Drug; D000939:Epitopes; D005353:Fibronectins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D039201:Integrin alpha3beta1; D019012:Integrin beta1 / Q000378:metabolism; D016023:Integrins / Q000737:chemistry* / Q000378:metabolism*; D007700:Kinetics; D008024:Ligands; D008545:Melanoma / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008954:Models, Biological; D010455:Peptides / Q000737:chemistry; D011485:Protein Binding; D011487:Protein Conformation; D015398:Signal Transduction; D019699:Thrombospondins / Q000737:chemistry / Q000378:metabolism*; D014407:Tumor Cells, Cultured
chemical_list: D000906:Antibodies; D000939:Epitopes; D005353:Fibronectins; D039201:Integrin alpha3beta1; D019012:Integrin beta1; D016023:Integrins; D008024:Ligands; D010455:Peptides; D019699:Thrombospondins; D002118:Calcium