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. 1993 May;91(5):1877–1883. doi: 10.1172/JCI116404

Importance of factor Xa in determining the procoagulant activity of whole-blood clots.

P R Eisenberg 1, J E Siegel 1, D R Abendschein 1, J P Miletich 1
PMCID: PMC288180  PMID: 8486759

Abstract

The binding of thrombin to fibrin is thought to be an important mechanism by which thrombi exhibit procoagulant activity; however, the extent to which other procoagulants are associated with thrombi has not been previously defined. This study was designed to determine whether clotting factors other than thrombin are bound to whole-blood clots and can thereby contribute to significant procoagulant activity. Clots formed in vitro from human blood exhibited minimal thrombin activity when incubated in plasma depleted of vitamin K-dependent factors by barium-citrate adsorption, as indicated by increases in the concentration of fibrinopeptide A (FPA), a marker of fibrin formation, to 72 nM after 30 min. Incubation of clots in barium-absorbed plasma repleted with 0.9 microM human prothrombin under the same conditions resulted in marked increases in the concentration of FPA (> 1,000 nM) and clotting by 30 min. The increases in FPA were attributable to activation of the added prothrombin by clot-associated Factor Xa, judging from concomitant increases in the concentration of prothrombin fragment 1.2. Similar results were obtained with thrombi induced in the axillary arteries of dogs by vascular injury and incubated with plasma in vitro. Activation of prothrombin was inhibited in a dose-dependent manner by tick anticoagulant peptide, a direct inhibitor of Factor Xa, at concentrations of 0.5-5.0 microM. Clot-associated Factor Xa activity was resistant to inhibition by anti-thrombin III, judging from the lack of inhibition of prothrombin activation during incubation of clots in plasma containing heparin pentasaccharide, an anti-thrombin III-mediated inhibitor of Factor Xa. Thus, the activity of Factor Xa appears to be an important determinant of the procoagulant activity of whole-blood clots and arterial thrombi, and is resistant to inhibition by anti-thrombin III-dependent inhibitors.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. CHANDLER A. B. In vitro thrombotic coagulation of the blood; a method for producing a thrombus. Lab Invest. 1958 Mar-Apr;7(2):110–114. [PubMed] [Google Scholar]
  2. Choay J., Lormeau J. C., Petitou M., Sinaÿ P., Fareed J. Structural studies on a biologically active hexasaccharide obtained from heparin. Ann N Y Acad Sci. 1981;370:644–649. doi: 10.1111/j.1749-6632.1981.tb29770.x. [DOI] [PubMed] [Google Scholar]
  3. Eisenberg P. R., Sherman L. A., Schectman K., Perez J., Sobel B. E., Jaffe A. S. Fibrinopeptide A: a marker of acute coronary thrombosis. Circulation. 1985 May;71(5):912–918. doi: 10.1161/01.cir.71.5.912. [DOI] [PubMed] [Google Scholar]
  4. Eisenberg P. R., Sobel B. E., Jaffe A. S. Activation of prothrombin accompanying thrombolysis with recombinant tissue-type plasminogen activator. J Am Coll Cardiol. 1992 Apr;19(5):1065–1069. doi: 10.1016/0735-1097(92)90296-y. [DOI] [PubMed] [Google Scholar]
  5. Francis C. W., Markham R. E., Jr, Barlow G. H., Florack T. M., Dobrzynski D. M., Marder V. J. Thrombin activity of fibrin thrombi and soluble plasmic derivatives. J Lab Clin Med. 1983 Aug;102(2):220–230. [PubMed] [Google Scholar]
  6. Hogg P. J., Jackson C. M. Fibrin monomer protects thrombin from inactivation by heparin-antithrombin III: implications for heparin efficacy. Proc Natl Acad Sci U S A. 1989 May;86(10):3619–3623. doi: 10.1073/pnas.86.10.3619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Jordan S. P., Waxman L., Smith D. E., Vlasuk G. P. Tick anticoagulant peptide: kinetic analysis of the recombinant inhibitor with blood coagulation factor Xa. Biochemistry. 1990 Dec 18;29(50):11095–11100. doi: 10.1021/bi00502a012. [DOI] [PubMed] [Google Scholar]
  8. Kettner C., Shaw E. D-Phe-Pro-ArgCH2C1-A selective affinity label for thrombin. Thromb Res. 1979;14(6):969–973. doi: 10.1016/0049-3848(79)90014-8. [DOI] [PubMed] [Google Scholar]
  9. Kockum C., Frebelius S. Rapid radioimmunoassay of human fibrinopeptide A - removal of cross-reacting fibrinogen with bentonite. 1980 Aug 15-Sep 1Thromb Res. 19(4-5):589–598. doi: 10.1016/0049-3848(80)90031-6. [DOI] [PubMed] [Google Scholar]
  10. Mann K. G., Nesheim M. E., Church W. R., Haley P., Krishnaswamy S. Surface-dependent reactions of the vitamin K-dependent enzyme complexes. Blood. 1990 Jul 1;76(1):1–16. [PubMed] [Google Scholar]
  11. Miletich J. P., Broze G. J., Jr, Majerus P. W. The synthesis of sulfated dextran beads for isolation of human plasma coagulation factors II, IX, and X. Anal Biochem. 1980 Jul 1;105(2):304–310. doi: 10.1016/0003-2697(80)90462-5. [DOI] [PubMed] [Google Scholar]
  12. Miletich J. P., Jackson C. M., Majerus P. W. Properties of the factor Xa binding site on human platelets. J Biol Chem. 1978 Oct 10;253(19):6908–6916. [PubMed] [Google Scholar]
  13. Neeper M. P., Waxman L., Smith D. E., Schulman C. A., Sardana M., Ellis R. W., Schaffer L. W., Siegl P. K., Vlasuk G. P. Characterization of recombinant tick anticoagulant peptide. A highly selective inhibitor of blood coagulation factor Xa. J Biol Chem. 1990 Oct 15;265(29):17746–17752. [PubMed] [Google Scholar]
  14. Nossel H. L., Yudelman I., Canfield R. E., Butler V. P., Jr, Spanondis K., Wilner G. D., Qureshi G. D. Measurement of fibrinopeptide A in human blood. J Clin Invest. 1974 Jul;54(1):43–53. doi: 10.1172/JCI107749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Schaffer L. W., Davidson J. T., Vlasuk G. P., Siegl P. K. Antithrombotic efficacy of recombinant tick anticoagulant peptide. A potent inhibitor of coagulation factor Xa in a primate model of arterial thrombosis. Circulation. 1991 Oct;84(4):1741–1748. doi: 10.1161/01.cir.84.4.1741. [DOI] [PubMed] [Google Scholar]
  16. Sitko G. R., Ramjit D. R., Stabilito I. I., Lehman D., Lynch J. J., Vlasuk G. P. Conjunctive enhancement of enzymatic thrombolysis and prevention of thrombotic reocclusion with the selective factor Xa inhibitor, tick anticoagulant peptide. Comparison to hirudin and heparin in a canine model of acute coronary artery thrombosis. Circulation. 1992 Feb;85(2):805–815. doi: 10.1161/01.cir.85.2.805. [DOI] [PubMed] [Google Scholar]
  17. Waxman L., Smith D. E., Arcuri K. E., Vlasuk G. P. Tick anticoagulant peptide (TAP) is a novel inhibitor of blood coagulation factor Xa. Science. 1990 May 4;248(4955):593–596. doi: 10.1126/science.2333510. [DOI] [PubMed] [Google Scholar]
  18. Weitz J. I., Hudoba M., Massel D., Maraganore J., Hirsh J. Clot-bound thrombin is protected from inhibition by heparin-antithrombin III but is susceptible to inactivation by antithrombin III-independent inhibitors. J Clin Invest. 1990 Aug;86(2):385–391. doi: 10.1172/JCI114723. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wilner G. D., Danitz M. P., Mudd M. S., Hsieh K. H., Fenton J. W., 2nd Selective immobilization of alpha-thrombin by surface-bound fibrin. J Lab Clin Med. 1981 Mar;97(3):403–411. [PubMed] [Google Scholar]

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