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Thromb Haemost 1995; 74(02): 698-703
DOI: 10.1055/s-0038-1649800
Original Article
Fibrinolysis
Schattauer GmbH Stuttgart

Assembly and Activation of the Intrinsic Fibrinolytic Pathway on the Surface of Human Endothelial Cells in Culture

Catherine Lenich
The Department of Medicine, Vascular Research Laboratory, Institute for the Prevention of Cardiovascular Disease, Deaconess Hospital, Harvard Medical School, Boston, USA
,
Ralph Pannell
The Department of Medicine, Vascular Research Laboratory, Institute for the Prevention of Cardiovascular Disease, Deaconess Hospital, Harvard Medical School, Boston, USA
,
Victor Gurewich
The Department of Medicine, Vascular Research Laboratory, Institute for the Prevention of Cardiovascular Disease, Deaconess Hospital, Harvard Medical School, Boston, USA
› Author Affiliations
Further Information

Publication History

Received 05 December 1994

Accepted after resubmission 04 April 1995

Publication Date:
06 July 2018 (online)

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Summary

Factor XII has long been implicated in the intrinsic pathway of fibrinolysis, but the mechanism by which it triggers plasminogen activation and targets fibrinolysis has not been established. In the present study, the assembly and function of activated Factor XII (F.XIIa), prourokinase (pro-u-PA), high molecular weight kininogen (H-kininogen), and prekallikrein on human umbilical vein endothelial cells (HUVEC) was investigated. 125I-prekallikrein was shown to bind to HUVEC via receptor-bound H-kininogen in the presence of 50 μM ZnCl2. After the addition of F.XIIa, 78% of the 125I-prekallikrein initially bound to HUVEC was converted to 125I-kallikrein. However, only 6% of the HUVEC-bound 125I-pro-u-PA was thereby activated. This discrepancy was shown to be related to rapid dissociation (>50% within 15 min) of prekallikrein/kallikrein, but not pro-u-PA, from HUVEC. Increasing the level of cell-bound kallikrein increased the portion of cell-bound pro-u-PA activated, indicating that their co-localization was important for this pathway. Finally, F.XIIa was shown to trigger plasminogen activation on HUVEC via this pathway. This assembly of reactants on the endothelium suggests a mechanism whereby local fibrinolysis may be triggered by blood coagulation.

 

  • References

  • 1 Booyse FM, Scheinbuks J, Lin PH, Traylor M, Bruce R. Isolation and interrelationships of the multiple molecular tissue-type and urokinase-type plasminogen activator forms produced by cultured human umbilical vein endothelial cells. J Biol Chem 1988; 263: 15129-15138
    PubMedSearch in Google Scholar
  • 2 Wun TC, Capuano A. Spontaneous fibrinolysis in whole plasma. Identification of tissue activator related protein as the major plasminogen activator causing spontaneous activity in vitro J Biol Chem 1985; 260: 5061-5066
    PubMedSearch in Google Scholar
  • 3 Collen D. On the regulation and control of fibrinolysis. Thromb Haemost 1980; 43: 77-89
    Thieme ConnectPubMedSearch in Google Scholar
  • 4 Kluft C, Dooijewaard G, Emeis JJ. Role of the contact system in fibrinolysis. Sem Thromb Haemost 1987; 13: 50-68
    Thieme ConnectPubMedSearch in Google Scholar
  • 5 Bamathan ES, Kuo A, Rosenfeld L, Kariko K, Leski M, Robbiati F, Nolli ML, Henkin J, Cines DB. Interaction of single-chain urokinase-type plasminogen activator with human endothelial cells. J Biol Chem 1990; 265: 2865-2872
    PubMedSearch in Google Scholar
  • 6 Haddock RC, Spell ML, Baker CD, Grammer JR, Parks JM, Speidel M, Booyse FM. Urokinase binding and receptor identification in cultured endothelial cells. J Biol Chem 1991; 266: 21466-21473
    PubMedSearch in Google Scholar
  • 7 Hajjar KA, Harpel PC, Jaffe EA, Nachman RL. Binding of plasminogen to cultured human endothelial cells. J Biol Chem 1986; 261: 11656-11562
    PubMedSearch in Google Scholar
  • 8 Ichinose A, Fujikawa K, Suyama T. The activation of pro-urokinase by plasma kallikrein and its inactivation by thrombin. J Biol Chem 1986; 261: 3486-3489
    PubMedSearch in Google Scholar
  • 9 Gurewich V, Pannell R. Inactivation of single-chain urokinase (pro-urokinase) by thrombin and thrombin-like enzymes: Relevance of the findings to the interpretation of fibrin-binding experiments. Blood 1987; 69: 769-772
    PubMedSearch in Google Scholar
  • 10 de Munk GA W, Groeneveld E, Rijken DC. Acceleration of the thrombin inactivation of single chain urokinase-type plasminogen activator (prourokinase) by thrombomodulin. J Clin Invest 1991; 88: 1680-1684
    CrossrefPubMedSearch in Google Scholar
  • 11 Loza J-P, Gurewich V, Johnstone M, Pannell R. Platelet-bound prekallikrein promotes pro-urokinase-induced clot lysis: a mechanism for targeting the factor XII dependent intrinsic pathway of fibrinolysis. Thromb Haemost 1994; 71: 347-352
    Thieme ConnectPubMedSearch in Google Scholar
  • 12 Castellino FJ, Powell JR. Human plasminogen. Methods Enzymol 1981; 80: 365-378
    PubMedSearch in Google Scholar
  • 13 Thompson RE, Mandle R, Kaplan AP. Studies of binding of prekallikrein and factor XI to high molecular weight kininogen and its light chain. Proc Natl Acad Sci USA 1979; 76: 4862-4866
    CrossrefPubMedSearch in Google Scholar
  • 14 van Iwaarden F, de Groot PG, Bouma BN. The binding of high molecular weight kininogen to cultured human endothelial cells. J Biol Chem 1988; 263: 4698-4703
    PubMedSearch in Google Scholar
  • 15 Schmaier AH, Kuo A, Lundberg D, Murray S, Cines DB. The expression of high molecular weight kininogen on human umbilical vein endothelial cells. J Biol Chem 1988; 263: 16327-16333
    PubMedSearch in Google Scholar
  • 16 Husain SS. Fibrin affinity of urokinase-type plasminogen activator. Evidence that Zn2+mediates strong and specific interaction of single-chain urokinase with fibrin J Biol Chem 1993; 268: 8574-8579
    PubMedSearch in Google Scholar
  • 17 Kitamura N, Kitagawa H, Fukushima D, Takagaki Y, Miyata T, Nakanishi S. Structural organization of the human kininogen gene and a model for its evolution. J Biol Chem 1985; 260: 8610-8617
    PubMedSearch in Google Scholar
  • 18 Reddigari SR, Shihayama Y, Brunnee T, Kaplan AP. Human llageman factor (factor XII) anti high molecular weight kininogen compelte for the same binding site on human umbilical vein endothelial cells. J Biol Clicm 1993; 268: 11982-11987
    PubMedSearch in Google Scholar
  • 19 Manehantda N, Sehwarlz BS. Single chain urokinase type plasminogen activator (SCU-PA) complexes with and is inhibited by plasminogen activator inhibitor type I (PAM-I). Fibrinolysis 1994; 8: 12 (Abstr.)
    PubMedSearch in Google Scholar
  • 20 Mandle Jr R, Colman RW, Kaplan AP. IdentiIicalion of prekallikrein and high molecular weight kininogen as a complex in human plasma. Proe Natl AeadSei 1976; 73: 4179-4178
    PubMedSearch in Google Scholar
  • 21 Bock PE, Shore JD. Protein-protein interactions in contact activation of blootl coagulation. J Biol Chem 1983; 238: 15079-15086
    PubMedSearch in Google Scholar
  • 22 Ellis V, Scully ME, Kakkar VV. Plasminogen activation initiated by singlechain urokinase-type plasminogen activator. Potentiation by U937 monocytes J Biol Chem 1989; 264: 2185-2188
    PubMedSearch in Google Scholar
  • 23 Heimark RL, Karachi K, Fujikawa K, Davie LW. Surface activation of blood coagulation, fibrinolysis and kinin formation. Nature 1980; 286: 456-460
    CrossrefPubMedSearch in Google Scholar
  • 24 Kaplan AP, Silverberg M. The eoagulalion-kinin pathway of human plasma. Blood 1987; 70: 1-15
    PubMedSearch in Google Scholar
  • 25 Goodnough LT, Saito H, Ratnoff OD. Thrombosis or myocardial infarction in congenital clotting factor abnormalities and chronic thrombocytopenias: A report of 21 patients and a review of 50 previously reported eases. Medicine (Baltimore) 1983 62: 248-255 70:1-15
    PubMed
  • 26 Miles LA, Greengard JS, Griffin JH. A comparison of the abilities ofplasma kallikrcin, factor XIIa, factor XIa and urokinase to activate plasminogen. Thomb Res 1983; 29: 407-417
    CrossrefPubMedSearch in Google Scholar
  • 27 Miles LA, Rothschild X, Griffin JH. Dcxtran sulphate-dependent fibrinolysis in whole human plasma. J Lab Clin Med 1983; 101: 214-225
    PubMedSearch in Google Scholar
  • 28 Huisveld IA, Hospers JE H, van Heeswijk GM, Berniek MJ E, Erich WB M, Bouma BN. Contribution of contact activation factors to urokinase-related fibrinolytic activity in whole human plasma. Thromb llaemost 1985; 54: 102 (Abstr.)
    PubMedSearch in Google Scholar