Thrombin Disintegrates Cell Surface Urokinase Focal Adhesion Plaques and Decreases Cell Extension: Implications for Axonal Growth

  • Caroline A. Hébert
  • Joffre B. Baker
Part of the NATO ASI Series book series (NSSA, volume 191)


In addition to its function as the ultimate protease in the coagulation cascade thrombin has several hormone-like effects on cells in vitro. This serine protease is a potent mitogen for fibroblastic cells from several species.1,2 In addition, like other growth factors,3 thrombin can modulate cellular differentiation. Thrombin or a closely related protease present on neuroblastoma cells antagonizes formation of neurite extensions.4 Furthermore thrombin has recently been identified as the primary serum inhibitor of neurite extension in neuroblastoma cell cultures.5 In the present paper we show that thrombin, but not other growth factors tested, causes the disappearance on human fibroblasts of linear strands of cell surface urokinase-type plasminogen activator, strands which have recently been shown to colocalize with cell-to-substratum focal adhesion plaques (FAPs)6,7 Disappearance or gross alteration of FAPs in fact accompanies the thrombin-mediated dissolution of the cell surfaced urokinase strands.


Platelet Derive Growth Factor Phorbol Myristate Acetate Phorbol Myristate Acetate Focal Contact Neurite Extension 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    D.H. Carney, K.C. Glenn, and D.D. Cunningham, Conditions which affect initiation of animal cell division by trypsin and thrombin. J. Cell. PhysioL, 95: 13–22 (1978).Google Scholar
  2. 2.
    L.B. Chen, and J.M. Buchanan, Mitogenic activity of blood components. I. Thrombin and prothrombin. Proc. NatL Acad. Sci USA 72: 131–135 (1975).Google Scholar
  3. 3.
    M.P. Sporn, and A.B. Roberts, Peptide growth factors are multifunctional. Nature 332: 217–219 (1988).CrossRefPubMedGoogle Scholar
  4. 4.
    D. Monard, E. Niday, A. Limat, and F. Solomon, Inhibition of protease activity can lead to neurite extension in neuroblastoma cells. Frog. Brain Res. 58: 359–364 (1983).Google Scholar
  5. 5.
    D. Gurwitz, and D D Cunningham, Thrombin modulates and reverses neuroblastoma neurite outgrowth. Proc. NatL Acad. Sci. USA 85: 3440–3444 (1988).CrossRefGoogle Scholar
  6. 6.
    CA. Hébert, and J.B. Baker, Linkage of extracellular plasminogen activator to the fibroblast cytoskeleton: colocalization of cell surface urokinase with vinculin. J. Cell BioL 106: 1241–1247 (1988).Google Scholar
  7. 7.
    J. Pöllänen, K. Hedman, L.S. Nielsen, K. Dan¢, and A. Vaheri, Ultrastructural localization of plasma membrane-associated urokinase-type plasminogen activator at focal contacts. J. Cell BioL 106: 87–95 (1988).Google Scholar
  8. 8.
    V.J. Hearing, L.W. Law, A. Corti, E. Appella, and F. Blasi, Modulation of metastatic potential by cell surface urokinase of murine melanoma cells. Cancer Res. 48: 1270–1278 (1988).PubMedGoogle Scholar
  9. 9.
    J.M. Vasiliev, Spreading of non-transformed and transformed cells. Biochim. Biophys. Acta 780: 21–65 (1985).Google Scholar
  10. 10.
    DA. Low, and D.D. Cunningham, A novel method for measuring cell surface-bound thrombin. J. BioL Chem. 257: 850–858 (1982).Google Scholar
  11. 11.
    D.L. Eaton, R.W. Scott, and J.B. Baker, Purification of human fibroblast urokinase proenzyme and analysis of its regulation by proteases and protease nexin. J. BioL Chem. 259: 6241–6247 (1984).Google Scholar
  12. 12.
    R.W. Scott, D.L. Eaton, N. Duran, and J.B. Baker, Regulation of extracellular plasminogen activator in human fibroblast cultures. The role of protease nexin. J. BioL Chem. 258: 4397–4403 (1983).Google Scholar
  13. 13.
    C.S. Izzard, and L.R. Lochner, Cell-to-substrate contacts in living fibroblasts: an interference reflection study with an evaluation of the technique. J. Cell Sci. 21: 129–159 (1976).Google Scholar
  14. 14.
    M.C. Willingham, and I. Pastan, An atlas of immunofluorescence in cultured cells. Academic Press, Inc., Orlando (1985).Google Scholar
  15. 15.
    F. Blasi, J.-D. Vassali, and K. DanO, Urokinase-type plasminogen activator: proenzyme, receptor, and inhibitors. J. Cell BioL 104: 801–804 (1987).Google Scholar
  16. 16.
    K. Dan¢, PA. Andreasen, J. Gr¢ndahl-Hansen, B. Kristensen, L.S. Nielsen, and L. Skriver, Plasminogen activators, tissue degradation and cancer. Adv. Cancer Res. 32: 146–239 (1985).Google Scholar
  17. 17.
    T. David-Pfeuty, and SJ. Singer, Altered distributions of the cytoskeletal protein vinculin and aactinin in cultured fibroblasts transformed by Rous sarcoma virus. Proc. Nat. Acad. Sd. USA 77: 6687–6691 (1980).Google Scholar
  18. D.L. Eaton, and J.B. Baker, Phorbol ester and mitogens stimulate human fibroblast secretions of plasmin-activatable plasminogen activator and protease nexin, an antiactivator/antiplasmin. J. Cell BioL7:323–328 (1983).Google Scholar
  19. 19.
    B. Herman, and WJ. Pledger, Platelet-derived growth factor-induced alterations in vinculin and actin distribution in BALB/c-3T3 cells. J. Cell BioL 100: 1031–1040 (1985).Google Scholar
  20. 20.
    B. Herman, MA. Harrington, N.E. Olashaw, and WJ. Pledger, Identification of the cellular mechanisms responsible for platelet-derived growth factor-induced alterations in cytoplasmic vinculin distribution. J. CelL PhysioL 126: 115–125 (1986).Google Scholar
  21. 21.
    DA. Low, R.W. Scott, J.B. Baker, and D.D. Cunningham, Cells regulate their mitogenic response to thrombin through secretion of protease nexin. Nature 298: 476–478 (1982).CrossRefPubMedGoogle Scholar
  22. 22.
    K.C. Glenn, D.H. Carney, J.W. Fenton, and D.D. Cunningham, Thrombin active site regions required for fibroblast receptor binding and initiation of cell division. J. BioL Chem. 225: 6609–6616 (1980).Google Scholar
  23. 23.
    A. Ichinose, K. Fujikawa, and T. Suyama, The activation of pro-urokinase by plasma kallikrein and its activation by thrombin. J. BioL Chem. 261: 3486–3489 (1986).Google Scholar
  24. 24.
    BA. Geiger, A 130K protein from chicken gizzard: its localization at the termini of microfilament bundles in cultured chicken cells. Cell 18: 193–205 (1979).CrossRefPubMedGoogle Scholar
  25. 25.
    D.M. Raben, K. Yasuda, and D.D. Cunningham, Thrombin stimulated metabolism of inositol-containing phospholipids. J. Celt Biochem. Suppl 0 (10 part A) p. 252.Google Scholar
  26. 26.
    N.H. Teng, and L.B. Chen, Thrombin-sensitive surface protein of cultured chick embryo cells. Nature 259: 578–580 (1976).CrossRefGoogle Scholar
  27. L.I. Gold, R. Schwimmer, and J.P. Quigley, Human plasma fibronectin is a substrate for the plasminogen activator, human urokinase. J. Cell BioL 105 p.216a (#1221) (1987).Google Scholar
  28. 28.
    M.-P. Stoppelli, C. Tacchetti, M.V. Cubellis, A. Corti, VJ. Hearing, G. Cassani, E. Appella, and F. Blasi, Autocrine saturation of prourokinase receptors on human A431 cells. Cell 45: 675–684 (1986).Google Scholar
  29. 29.
    J. Kolega, M.S. Shure, W.-T. Chen, and N.D. Young, Rapid cellular translocation is related to close contacts formed between various cultured cells and their substrata. J. Cell Sci. 54: 23–34 (1982).Google Scholar
  30. 30.
    K. Burridge, Substrate adhesions in normal and transformed fibroblasts: organization and regulation of cytoskeletal, membrane and extracellular matrix components at focal contacts. Cancer Rev. 4: 18–78 (1986).Google Scholar
  31. 31.
    D. Schubert, S. Humphreys, C. Baroni, and M. Cohn, In vitro differentiation of a mouse neuroblastoma. Proc. Natl. Acad. Sci. USA 64: 316–323 (1969).CrossRefPubMedGoogle Scholar
  32. 32.
    N.W. Seeds, A.G. Gilman, T. Amano, and M.W. Niremberg, Regulation of axon formation by clonal lines of a neuronal tumor. Proc. Natl. Acad. Sci USA 66: 160–167 (1970).CrossRefPubMedGoogle Scholar
  33. 33.
    B. Geiger, Z. Avnur, G. Rinnerthaler, H. Hinssen, and J.V. Small, Microfilament-organizing centers in areas of cell contact: cytoskeletal interactions during cell attachment and locomotion. J. Cell BioL 99: 83–91 (1984).CrossRefGoogle Scholar
  34. 34.
    J.V. Small, and G. Rinnerthaler, Cytostructural dynamics of contact formation during fibroblast locomotion in vitro. Exp. Bio. Med. 10: 54–68 (1985).Google Scholar
  35. 35.
    S. Halegoua, Changes in phosphorylation and distribution of vinculin during nerve growth factor-induced neurite outgrowth. Dev. BioL 121: 97–104 (1987).Google Scholar
  36. 36.
    J.-D. Vassalli, D. Baccino, and D. Belin, A cellular binding site for the Mr. 55,000 form of the human plasminogen activator, urokinase. J. Cell BioL 100: 86–92 (1985).Google Scholar

Copyright information

© Plenum Press, New York 1990

Authors and Affiliations

  • Caroline A. Hébert
    • 1
  • Joffre B. Baker
    • 1
  1. 1.Department of BiochemistryUniversity of KansasLawrenceUSA

Personalised recommendations