Identification of a urokinase receptor-integrin interaction site. Promiscuous regulator of integrin function.
Adhesion and signaling by integrins require their dynamic association with nonintegrin membrane proteins. One such protein, the glycolipid-anchored urokinase receptor (uPAR), associates with and modifies the function of the beta(2)-integrin Mac-1 (CD11b/CD18). In this study, a critical non-I-domain binding site for uPAR on CD11b (M25; residues 424-440) is identified by homology with a phage display peptide known to bind uPAR. Recombinant soluble uPAR and cells expressing uPAR bound to immobilized M25, binding being promoted by urokinase and blocked by soluble M25, but not a scrambled control or homologous peptides from other beta(2)-associated alpha-chains. Mac-1, but not a mutated Mac-1 in which M25 was replaced with the homologous sequence of CD11c, co-precipitated with uPAR. In the beta-propeller model of alpha-chain folding, M25 spans an exposed loop on the ligand-binding, upper surface of alphaM, identifying uPAR as an atypical alphaM ligand. Although not blocking ligand binding to Mac-1, M25 (25-100 microM) inhibited leukocyte adhesion to fibrinogen, vitronectin, and cytokine-stimulated endothelial cells. M25 also blocked the association of uPAR with beta(1)-integrins and impaired beta(1)-integrin-dependent spreading and migration of human vascular smooth muscle cells on fibronectin and collagen. These observations indicate that uPAR associates with integrins directly and that disruption of this association broadly impairs integrin function, suggesting a novel strategy for regulation of integrins in the settings of inflammation and tumor progression.
Simon
D I
DI
Cardiovascular, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Wei
Y
Y
Zhang
L
L
Rao
N K
NK
Xu
H
H
Chen
Z
Z
Liu
Q
Q
Rosenberg
S
S
Chapman
H A
HA
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000737:chemistry / Q000378:metabolism; D001665:Binding Sites; D018821:CD18 Antigens / Q000737:chemistry* / Q000378:metabolism*; D002460:Cell Line; D002633:Chemotaxis; D006801:Humans; D046911:Macromolecular Substances; D016177:Macrophage-1 Antigen / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology*; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D010958:Plasminogen / Q000378:metabolism; D010960:Plasminogen Activators / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000378:metabolism*; D055293:Receptors, Urokinase Plasminogen Activator; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D012501:Saphenous Vein / Q000166:cytology / Q000502:physiology; D014162:Transfection
D015703:Antigens, CD; D018821:CD18 Antigens; D046911:Macromolecular Substances; D016177:Macrophage-1 Antigen; C524898:PLAUR protein, human; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011993:Recombinant Fusion Proteins; D010958:Plasminogen; D010960:Plasminogen Activators