title: Kinetic analysis of the interaction between vitronectin and the urokinase receptor.
abstract: Although the urokinase receptor (uPAR) binds to vitronectin (VN) and promotes the adhesion of cells to this matrix protein, the biochemical details of this interaction remain unclear. VN variants were employed in BIAcore experiments to examine the uPAR-VN interaction in detail and to compare it to the interaction of VN with other ligands. Heparin and plasminogen bound to VN fragments containing the heparin-binding domain, indicating that this domain was functionally active in the recombinant peptides. However, no significant binding was detected when uPAR was incubated with this domain, and neither heparin nor plasminogen competed with it for binding to VN. In fact, uPAR only bound to fragments containing the somatomedin B (SMB) domain, and monoclonal antibodies (mAbs) that bind to this domain competed with uPAR for binding to VN. Monoclonal antibody 8E6 also inhibited uPAR binding to VN, and this mAb was shown to recognize sulfated tyrosine residues 56 and 59 in the region adjacent to the SMB domain. Destruction of this site by acid treatment eliminated mAb 8E6 binding but had no effect on uPAR binding. Thus, there appears to be a single binding site for uPAR in VN, and it is located in the SMB domain and is distinct from the epitope recognized by mAb 8E6. Inhibition of uPAR binding to VN by mAb 8E6 probably results from steric hindrance.
authors.affiliation: Department of Cell Biology, Division of Vascular Biology, Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Yuushi
authors.initials: Y
authors.lastname: Okumura
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Kamikubo
authors.forename: Scott A
authors.initials: SA
authors.lastname: Curriden
authors.forename: Jieyi
authors.initials: J
authors.lastname: Wang
authors.forename: Tatsuto
authors.initials: T
authors.lastname: Kiwada
authors.forename: Shiroh
authors.initials: S
authors.lastname: Futaki
authors.forename: Kouki
authors.initials: K
authors.lastname: Kitagawa
authors.forename: David J
authors.initials: DJ
authors.lastname: Loskutoff
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D002448:Cell Adhesion; D000939:Epitopes; D006801:Humans; D007700:Kinetics; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D011489:Protein Denaturation; D011956:Receptors, Cell Surface / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator; D013002:Somatomedins / Q000378:metabolism; D020298:U937 Cells; D019096:Vitronectin / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; C524898:PLAUR protein, human; D017395:Plasminogen Activator Inhibitor 1; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D013002:Somatomedins; D019096:Vitronectin; C016756:somatomedin B