Direct interaction of the kringle domain of urokinase-type plasminogen activator (uPA) and integrin αvβ3 induces signal transduction and enhances plasminogen activation

 

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Summary

It has been questioned whether there are receptors for urokinase-type plasminogen activator (uPA) that facilitate plasminogen activation other than the high affinity uPA receptor (uPAR/CD87) since studies of uPAR knockout mice did not support a major role of uPAR in plasminogen activation. uPA also promotes cell adhesion, chemotaxis, and proliferation besides plasminogen activation. These uPA-induced signaling events are not mediated by uPAR,but mediated by unidentified,lower-affinity receptors for the uPA kringle.We found that uPA binds specifically to integrin αvβ3 on CHO cells depleted of uPAR. The binding of uPA to αvβ3 required the uPA kringle domain. The isolated uPA kringle domain binds specifically to purified,recombinant soluble, and cell surface αvβ3, and other integrins (α4β1 and α9β1), and induced migration of CHO cells in an αvβ3-dependent manner. The binding of the uPA kringle to αvβ3 and uPA kringle-induced αvβ3-dependent cell migration were blocked by homologous plasminogen kringles 1-3 or 1-4 (angiostatin),a known integrin antagonist. We studied whether the binding of uPA to integrin αvβ3 through the kringle domain plays a role in plasminogen activation. On CHO cell depleted of uPAR, uPA enhanced plasminogen activation in a kringle and αvβ3-dependent manner.Endothelial cells bound to and migrated on uPA and uPA kringle in an αvβ3-dependent manner. These results suggest that uPA binding to integrins through the kringle domain plays an important role in both plasminogen activation and uPA-induced intracellular signaling. The uPA kringle-integrin interaction may represent a novel therapeutic target for cancer, inflammation, and vascular remodeling.

• References

• 1 Chapman HA. Plasminogen activators, integrins, and the coordinated regulation of cell adhesion and migration. Curr Opin Cell Biol 1997; 09: 714-24.
  CrossrefPubMedGoogle Scholar
• 2 Andreasen PA, Kjoller L, Christensen L. et al. The urokinase-type plasminogen activator system in cancer metastasis: a review. Int J Cancer 1997; 72: 1-22.
  CrossrefPubMedGoogle Scholar
• 3 Dano K, Romer J, Nielsen BS. et al. Cancer invasion and tissue remodeling--cooperation of protease systems and cell types. Apmis 1999; 107: 120-7.
  CrossrefPubMedGoogle Scholar
• 4 Mondino A, Blasi F. uPA and uPAR in fibrinolysis, immunity and pathology. Trends Immunol 2004; 25: 450-5.
  CrossrefPubMedGoogle Scholar
• 5 Appella E, Blasi F. The growth factor module of urokinase is the binding sequence for its receptor. Ann NY Acad Sci 1987; 511: 192-5.
  CrossrefPubMedGoogle Scholar
• 6 Bdeir K, Kuo A, Sachais BS. et al. The kringle stabilizes urokinase binding to the urokinase receptor. Blood 2003; 102: 3600-8.
  CrossrefPubMedGoogle Scholar
• 7 Waltz DA, Fujita RM, Yang X. et al. Nonproteolytic role for the urokinase receptor in cellular migration in vivo . AmJ Respir Cell Mol Biol 2000; 22: 316-22.
  PubMedGoogle Scholar
• 8 Ellis V, Pyke C, Eriksen J. et al. The urokinase receptor: involvement in cell surface proteolysis and cancer invasion. Ann NY Acad Sci 1992; 667: 13-31.
  CrossrefPubMedGoogle Scholar
• 9 Carmeliet P, Schoonjans L, Kieckens L. et al. Physiological consequences of loss of plasminogen activator gene function in mice. 1994; 368: 419-24.
  PubMedGoogle Scholar
• 10 Bugge TH, Flick MJ, Danton MJ. et al. Urokinasetype plasminogen activator is effective in fibrin clearance in the absence of its receptor or tissue-type plasminogen activator. Proc Natl Acad Sci USA 1996; 93: 5899-904.
  CrossrefPubMedGoogle Scholar
• 11 Waltz DA, Sailor LZ, Chapman HA. Cytokines induce urokinase-dependent adhesion of human myeloid cells A regulatory role for plasminogen activator inhibitors. J Clin Invest 1993; 91: 1541-52.
  CrossrefPubMedGoogle Scholar
• 12 Gyetko MR, Todd 3rd RF, Wilkinson CC. et al. The urokinase receptor is required for human monocyte chemotaxis in vitro . J Clin Invest 1994; 93: 1380-7.
  CrossrefPubMedGoogle Scholar
• 13 Busso N, Masur SK, Lazega D. et al. Induction of cell migration by pro-urokinase binding to its receptor: possible mechanism for signal transduction in human epithelial cells. J Cell Biol 1994; 126: 259-70.
  CrossrefPubMedGoogle Scholar
• 14 Odekon LE, Sato Y, Rifkin DB. Urokinase-type plasminogen activator mediates basic fibroblast growth factor-induced bovine endothelial cell migration independent of its proteolytic activity. J Cell Physiol 1992; 150: 258-63.
  CrossrefPubMedGoogle Scholar
• 15 Rabbani SA, Mazar AP, Bernier SM. et al. Structural requirements for the growth factor activity of the amino-terminal domain of urokinase. J Biol Chem 1992; 267: 14151-6.
  PubMedGoogle Scholar
• 16 Aguirre Ghiso JA, Kovalski K, Ossowski L. Tumor dormancy induced by downregulation of urokinase receptor in human carcinoma involves integrin and MAPK signaling. J Cell Biol 1999; 147: 89-104.
  CrossrefPubMedGoogle Scholar
• 17 Fischer K, Lutz V, Wilhelm O. et al. Urokinase induces proliferation of human ovarian cancer cells: characterization of structural elements required for growth factor function. FEBS Lett 1998; 438: 101-5.
  CrossrefPubMedGoogle Scholar
• 18 Kanse SM, Benzakour O, Kanthou C. et al. Induction of vascular SMC proliferation by urokinase indicates a novel mechanism of action in vasoproliferative disorders. Arterioscler Thromb Vasc Biol 1997; 17: 2848-54.
  CrossrefPubMedGoogle Scholar
• 19 Koopman JL, Slomp J, de Bart AC. et al. Mitogenic effects of urokinase on melanoma cells are independent of high affinity binding to the urokinase receptor. J Biol Chem 1998; 273: 33267-72.
  CrossrefPubMedGoogle Scholar
• 20 Mukhina S, Stepanova V, Traktouev D. et al. The chemotactic action of urokinase on smooth muscle cells is dependent on its kringle domain; Characterization of interactions and contribution to chemotaxis. J Biol Chem 2000; 275: 16450-8.
  CrossrefPubMedGoogle Scholar
• 21 Haj-Yehia A, Nassar T, Sachais BS. et al. Urokinase-derived peptides regulate vascular smooth muscle contraction in vitro and in vivo . Faseb J 2000; 14: 1411-22.
  CrossrefPubMedGoogle Scholar
• 22 Nassar T, Haj-Yehia A, Akkawi S. et al. Binding of urokinase to low density lipoprotein-related receptor (LRP) regulates vascular smooth muscle cell contraction. J Biol Chem 2002; 277: 40499-504.
  CrossrefPubMedGoogle Scholar
• 23 Miyake K, Hasunuma Y, Yagita H. et al. Requirement for VLA-4 and VLA-5 integrins in lymphoma cells binding to and migration beneath stromal cells in culture. J Cell Biol 1992; 119: 653-62.
  CrossrefPubMedGoogle Scholar
• 24 Wang A, Yokosaki Y, Ferrando R. et al. Differential regulation of airway epithelial integrins by growth factors. Am J Respir Cell Mol Biol 1996; 15: 664-72.
  CrossrefPubMedGoogle Scholar
• 25 Coller BS. A new murine monoclonal antibody reports an activation-dependent change in the conformation and/or microenvironment of the platelet glycoprotein IIb/IIIa complex. J Clin Invest 1985; 76: 101-8.
  CrossrefPubMedGoogle Scholar
• 26 Deckmyn H, Stanssens P, Hoet B. et al. An echistatin-like Arg-Gly-Asp (RGD)-containing sequence in the heavy chain CDR3 of a murine monoclonal antibody that inhibits human platelet glycoprotein IIb/IIIa function. Br J Haematol 1994; 87: 562-71.
  CrossrefPubMedGoogle Scholar
• 27 Cheresh DA, Smith JW, Cooper HM. et al. A novel vitronectin receptor integrin (avbx) is responsible for distinct adhesive properties of carcinoma cells. Cell 1989; 57: 59-69.
  CrossrefPubMedGoogle Scholar
• 28 Min HY, Semnani R, Mizukami IF. et al. cDNA for Mo3, a monocyte activation antigen, encodes the human receptor for urokinase plasminogen activator. J Immunol 1992; 148: 3636-42.
  PubMedGoogle Scholar
• 29 Higazi AAR, Mazar A, Wang J. et al. Soluble human urokinase receptor is composed of two active units. J Biol Chem 1997; 272: 5348-53.
  CrossrefPubMedGoogle Scholar
• 30 Takagi J, DeBottis DP, Erickson HP. et al. The role of the specificity-determining loop of the integrin beta subunit I-like domain in autonomous expression, association with the alpha subunit, and ligand binding. Biochemistry 2002; 41: 4339-47.
  CrossrefPubMedGoogle Scholar
• 31 Zhang XP, Kamata T, Yokoyama K. et al. Specific interaction of the recombinant disintegrin-like domain of MDC- 15 (metargidin, ADAM-15) with integrin αvβ3. J Biol Chem 1998; 273: 7345-50.
  CrossrefPubMedGoogle Scholar
• 32 Tarui T, Mazar AP, Cines DB. et al. Urokinase-type plasminogen activator receptor (CD87) is a ligand for integrins and mediates cell-cell interaction. J Biol Chem 2001; 276: 3983-90.
  CrossrefPubMedGoogle Scholar
• 33 Schreiner CL, Bauer JS, Danilov YN. et al. Isolation and characterization of chinese hamster ovary cell variants deficient in the expression of fibronectin receptor. J Cell Biol 1989; 109: 3157-67.
  CrossrefPubMedGoogle Scholar
• 34 Prater CA, Plotkin J, Jaye D. et al. The properdinlike type I repeats of human thrombospondin containa cell attachment site. J Cell Biol 1991; 112: 1031-40.
  CrossrefPubMedGoogle Scholar
• 35 Takagi J, Erickson HP, Springer TA. C-terminal opening mimics ‘inside-out’ activation of integrin α5β1. Nat Struct Biol 2001; 08: 412-6.
  CrossrefPubMedGoogle Scholar
• 36 Takada Y, Puzon W. Identification of a regulatory region of integrin b1 subunit using activating and inhibiting antibodies. J Biol Chem 1993; 268: 17597-601.
  PubMedGoogle Scholar
• 37 Tarui T, Andronicos N, Czekay RP. et al. Critical role of integrin α5β1 in urokinase (uPA)/urokinase receptor (uPAR, CD87) signaling. J Biol Chem 2003; 278: 29863-72.
  CrossrefPubMedGoogle Scholar
• 38 Tarui T, Miles LA, Takada Y. Specific interaction of angiostatin with integrin αvβ3 in endothelial cells. J Biol Chem 2001; 276: 39562-8.
  CrossrefPubMedGoogle Scholar
• 39 Tarui T, Majumdar M, Miles LA. et al. Plasmin-induced migration of endothelial cells:A potential target for the anti-angiogenic action of angiostatin. J Biol Chem 2002; 277: 33564-70.
  CrossrefPubMedGoogle Scholar
• 40 Ugarova T, Budzynski A, Shattil S. et al. Conformational changes in fibrinogen elicited by its interaction with platelet membrane glycoprotein GPIIb-IIIa. J Biol Chem 1993; 268: 21080-7.
  PubMedGoogle Scholar
• 41 Seiffert D, Smith JW. The cell adhesion domain in plasma vitronectin is cryptic. J Biol Chem 1997; 272: 13705-10.
  CrossrefPubMedGoogle Scholar
• 42 Eliceiri BP, Cheresh DA. The role of alphav integrins during angiogenesis: insights into potential mechanisms of action and clinical development. J Clin Invest 1999; 103: 1227-30.
  CrossrefPubMedGoogle Scholar
• 43 Eliceiri BP. Integrin and growth factor receptor crosstalk. Circ Res 2001; 89: 1104-10.
  CrossrefPubMedGoogle Scholar
• 44 Kwak S-H, Mitra S, Bdeir K. et al. The kringle domain of urokinase-type plasminogen activator potentiates LPS-induced neutrophil activation through interaction with αvβ3 integrins. J Leukoc Biol 2005; 78: 937-45.
  CrossrefPubMedGoogle Scholar
• 45 Ugarova TP, Zamarron C, Veklich Y. et al. Conformational transitions in the cell binding domain offibronectin. Biochemistry 1995; 34: 4457-66.
  CrossrefPubMedGoogle Scholar
• 46 Pluskota E, Soloviev DA, Plow EF. Convergence of the adhesive and fibrinolytic systems: recognition of urokinase by integrin αMβ2 as well as by the urokinase receptor regulates cell adhesion and migration. Blood 2003; 101: 1582-90.
  CrossrefPubMedGoogle Scholar
• 47 Pluskota E, Soloviev DA, Bdeir K. et al. Integrin aMß2 orchestrates and accelerates plasminogen activation and fibrinolysis by neutrophils. J Biol Chem 2004; 279: 18063-72.
  CrossrefPubMedGoogle Scholar
• 48 Celikel R. Crystallization of the OP-G2 Fab fragment: a fibrinogen mimic with specificity for the platelet glycoprotein IIb/IIIa. Acta Crystallogr D Biol Crystallogr 1993; 49: 421-2.
  CrossrefPubMedGoogle Scholar
• 49 Miao H, Li S, Hu YL. et al. Differential regulation of Rho GTPases by beta1 and beta3 integrins: the role of an extracellular domain of integrin in intracellular signaling. J Cell Sci 2002; 11: 2199-206.
  PubMedGoogle Scholar
• 50 Ossowski L, Aguirre-Ghiso JA. Urokinase receptor and integrin partnership: coordination of signaling for cell adhesion, migration and growth. Curr Opin Cell Biol 2000; 12: 613-20.
  CrossrefPubMedGoogle Scholar