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. 1988 Nov;82(5):1700–1707. doi: 10.1172/JCI113783

Human tissue-type plasminogen activator releases fibrinopeptides A and B from fibrinogen.

J I Weitz 1, M K Cruickshank 1, B Thong 1, B Leslie 1, M N Levine 1, J Ginsberg 1, T Eckhardt 1
PMCID: PMC442740  PMID: 3141481

Abstract

In five patients with venous thromboembolic disease treated with recombinant tissue-type plasminogen activator (rt-PA), there was a marked increase in the mean concentrations of fibrinopeptide A (from 0.6 to 5.9 nM; P less than 0.0001) and desarginine fibrinopeptide B (from 5.6 nM to 24.1 nM; P less than 0.01) 30 min after a bolus of rt-PA (0.6 mg/kg). Thrombin was unlikely to be responsible because the levels of desarginine fibrinopeptide B exceeded those of fibrinopeptide A and the changes occurred despite concomitant heparin therapy. The purpose of this study therefore, was to determine whether rt-PA directly releases the fibrinopeptides from fibrinogen. Incubation of rt-PA with heparinized plasma or purified fibrinogen resulted in time and dose-dependent release of both fibrinopeptide A and B. Contaminating thrombin was not responsible for this activity by the following criteria: the rate of rt-PA mediated fibrinopeptide B release was considerably faster than that of fibrinopeptide A, and fibrinopeptide release was unaffected by heparin, hirudin, or a monospecific antithrombin IgG. Aprotinin also had no effect on fibrinopeptide release, indicating that this activity was not plasmin mediated. Fibrinopeptide release was shown to be due to rt-PA because this activity was completely blocked by a monoclonal antibody against the enzyme. Further, the specificity of rt-PA for the thrombin cleavage sites on fibrinogen was confirmed by the demonstration that rt-PA released fibrinopeptide A or fibrinopeptide B from fibrinopeptide A or B-containing substrates, respectively. These studies thus demonstrate that (a) rt-PA releases fibrinopeptides A and B from fibrinogen thereby indicating that this enzyme is not specific for plasminogen, and (b) plasma fibrinopeptide A and desarginine fibrinopeptide B levels are not specific markers of thrombin action on fibrinogen in patients receiving rt-PA.

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Selected References

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

  1. Bilezikian S. B., Nossel H. L., Butler V. P., Jr, Canfield R. E. Radioimmunoassay of human fibrinopeptide B and kinetics of fibrinopeptide cleavage by different enzymes. J Clin Invest. 1975 Aug;56(2):438–445. doi: 10.1172/JCI108110. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Canfield R. E., Dean J., Nossel H. L., Butler V. P., Jr, Wilner G. D. Reactivity of fibrinogen and fibrinopeptide A containing fibrinogen fragments with antisera to fibrinopeptide A. Biochemistry. 1976 Mar 23;15(6):1203–1209. doi: 10.1021/bi00651a004. [DOI] [PubMed] [Google Scholar]
  3. Chesebro J. H., Knatterud G., Roberts R., Borer J., Cohen L. S., Dalen J., Dodge H. T., Francis C. K., Hillis D., Ludbrook P. Thrombolysis in Myocardial Infarction (TIMI) Trial, Phase I: A comparison between intravenous tissue plasminogen activator and intravenous streptokinase. Clinical findings through hospital discharge. Circulation. 1987 Jul;76(1):142–154. doi: 10.1161/01.cir.76.1.142. [DOI] [PubMed] [Google Scholar]
  4. Chibber B. A., Deutsch D. G., Mertz E. T. Affinity chromatography of plasminogen. Methods Enzymol. 1974;34:424–432. doi: 10.1016/s0076-6879(74)34051-7. [DOI] [PubMed] [Google Scholar]
  5. Collen D., Topol E. J., Tiefenbrunn A. J., Gold H. K., Weisfeldt M. L., Sobel B. E., Leinbach R. C., Brinker J. A., Ludbrook P. A., Yasuda I. Coronary thrombolysis with recombinant human tissue-type plasminogen activator: a prospective, randomized, placebo-controlled trial. Circulation. 1984 Dec;70(6):1012–1017. doi: 10.1161/01.cir.70.6.1012. [DOI] [PubMed] [Google Scholar]
  6. Eckhardt T., Nossel H. L., Hurlet-Jensen A., La Gamma K. S., Owen J., Auerbach M. Measurement of desarginine fibrinopeptide B in human blood. J Clin Invest. 1981 Mar;67(3):809–816. doi: 10.1172/JCI110098. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Eisenberg P. R., Sherman L., Rich M., Schwartz D., Schechtman K., Geltman E. M., Sobel B. E., Jaffe A. S. Importance of continued activation of thrombin reflected by fibrinopeptide A to the efficacy of thrombolysis. J Am Coll Cardiol. 1986 Jun;7(6):1255–1262. doi: 10.1016/s0735-1097(86)80144-9. [DOI] [PubMed] [Google Scholar]
  8. Gold H. K., Leinbach R. C., Garabedian H. D., Yasuda T., Johns J. A., Grossbard E. B., Palacios I., Collen D. Acute coronary reocclusion after thrombolysis with recombinant human tissue-type plasminogen activator: prevention by a maintenance infusion. Circulation. 1986 Feb;73(2):347–352. doi: 10.1161/01.cir.73.2.347. [DOI] [PubMed] [Google Scholar]
  9. Hajjar K. A., Hamel N. M., Harpel P. C., Nachman R. L. Binding of tissue plasminogen activator to cultured human endothelial cells. J Clin Invest. 1987 Dec;80(6):1712–1719. doi: 10.1172/JCI113262. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harrison D. G., Ferguson D. W., Collins S. M., Skorton D. J., Ericksen E. E., Kioschos J. M., Marcus M. L., White C. W. Rethrombosis after reperfusion with streptokinase: importance of geometry of residual lesions. Circulation. 1984 May;69(5):991–999. doi: 10.1161/01.cir.69.5.991. [DOI] [PubMed] [Google Scholar]
  11. Herzig R. H., Ratnoff O. D., Shainoff J. R. Studies on a procoagulant fraction of southern copperhead snake venom: the preferential release of fibrinopeptide B. J Lab Clin Med. 1970 Sep;76(3):451–465. [PubMed] [Google Scholar]
  12. Hoylaerts M., Rijken D. C., Lijnen H. R., Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin. J Biol Chem. 1982 Mar 25;257(6):2912–2919. [PubMed] [Google Scholar]
  13. 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]
  14. Kluft C. Studies on the fibrinolytic system in human plasma: quantitative determination of plasminogen activators and proactivators. Thromb Haemost. 1979 Apr 23;41(2):365–383. [PubMed] [Google Scholar]
  15. La Gamma K. S., Nossel H. L. The stability of fibrinopeptide B immunoreactivity in blood. Thromb Res. 1978 Mar;12(3):447–454. doi: 10.1016/0049-3848(78)90315-8. [DOI] [PubMed] [Google Scholar]
  16. Levin E. G., Marzec U., Anderson J., Harker L. A. Thrombin stimulates tissue plasminogen activator release from cultured human endothelial cells. J Clin Invest. 1984 Dec;74(6):1988–1995. doi: 10.1172/JCI111620. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Loskutoff D. J. Effect of thrombin on the fibrinolytic activity of cultured bovine endothelial cells. J Clin Invest. 1979 Jul;64(1):329–332. doi: 10.1172/JCI109457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. MacGregor I. R., Micklem L. R., James K., Pepper D. S. Characterisation of epitopes on human tissue plasminogen activator recognised by a group of monoclonal antibodies. Thromb Haemost. 1985 Feb 18;53(1):45–50. [PubMed] [Google Scholar]
  19. Marglin A., Merrifield R. B. Chemical synthesis of peptides and proteins. Annu Rev Biochem. 1970;39:841–866. doi: 10.1146/annurev.bi.39.070170.004205. [DOI] [PubMed] [Google Scholar]
  20. Nilsson T., Wallén P., Mellbring G. In vivo metabolism of human tissue-type plasminogen activator. Scand J Haematol. 1984 Jul;33(1):49–53. doi: 10.1111/j.1600-0609.1984.tb02209.x. [DOI] [PubMed] [Google Scholar]
  21. Noe D. A., Bell W. R. A kinetic analysis of fibrinogenolysis during plasminogen activator therapy. Clin Pharmacol Ther. 1987 Mar;41(3):297–303. doi: 10.1038/clpt.1987.31. [DOI] [PubMed] [Google Scholar]
  22. Nossel H. L., Butler V. P., Jr, Wilner G. D., Canfield R. E., Harfenist E. J. Specificity of antisera to human fibrinopeptide A used in clinical fibrinopeptide A assays. Thromb Haemost. 1976 Feb 29;35(1):101–109. [PubMed] [Google Scholar]
  23. 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]
  24. Owen J., Friedman K. D., Grossman B. A., Wilkins C., Berke A. D., Powers E. R. Quantitation of fragment X formation during thrombolytic therapy with streptokinase and tissue plasminogen activator. J Clin Invest. 1987 Jun;79(6):1642–1647. doi: 10.1172/JCI113001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Owen J., Friedman K. D., Grossman B. A., Wilkins C., Berke A. D., Powers E. R. Thrombolytic therapy with tissue plasminogen activator or streptokinase induces transient thrombin activity. Blood. 1988 Aug;72(2):616–620. [PubMed] [Google Scholar]
  26. Peuscher F. W., van Aken W. G., Flier O. T., Stoepman-van Dalen E. A., Cremer-Goote T. M., van Mourik J. A. Effect of anticoagulant treatment measured by fibrinopeptide A (fpA) in patients with venous thrombo-embolism. Thromb Res. 1980 Apr 1;18(1-2):33–43. doi: 10.1016/0049-3848(80)90168-1. [DOI] [PubMed] [Google Scholar]
  27. Plow E. F. Leukocyte elastase release during blood coagulation. A potential mechanism for activation of the alternative fibrinolytic pathway. J Clin Invest. 1982 Mar;69(3):564–572. doi: 10.1172/JCI110482. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rijken D. C., Collen D. Purification and characterization of the plasminogen activator secreted by human melanoma cells in culture. J Biol Chem. 1981 Jul 10;256(13):7035–7041. [PubMed] [Google Scholar]
  29. Rijken D. C., Hoylaerts M., Collen D. Fibrinolytic properties of one-chain and two-chain human extrinsic (tissue-type) plasminogen activator. J Biol Chem. 1982 Mar 25;257(6):2920–2925. [PubMed] [Google Scholar]
  30. Rijken D. C., Hoylaerts M., Collen D. Fibrinolytic properties of one-chain and two-chain human extrinsic (tissue-type) plasminogen activator. J Biol Chem. 1982 Mar 25;257(6):2920–2925. [PubMed] [Google Scholar]
  31. Rånby M. Studies on the kinetics of plasminogen activation by tissue plasminogen activator. Biochim Biophys Acta. 1982 Jun 24;704(3):461–469. doi: 10.1016/0167-4838(82)90068-1. [DOI] [PubMed] [Google Scholar]
  32. Sobel B. E., Gross R. W., Robison A. K. Thrombolysis, clot selectivity, and kinetics. Circulation. 1984 Aug;70(2):160–164. doi: 10.1161/01.cir.70.2.160. [DOI] [PubMed] [Google Scholar]
  33. Topol E. J., Bell W. R., Weisfeldt M. L. Coronary thrombolysis with recombinant tissue-type plasminogen activator. A hematologic and pharmacologic study. Ann Intern Med. 1985 Dec;103(6 ):837–843. doi: 10.7326/0003-4819-103-6-837. [DOI] [PubMed] [Google Scholar]
  34. Van de Werf F., Ludbrook P. A., Bergmann S. R., Tiefenbrunn A. J., Fox K. A., de Geest H., Verstraete M., Collen D., Sobel B. E. Coronary thrombolysis with tissue-type plasminogen activator in patients with evolving myocardial infarction. N Engl J Med. 1984 Mar 8;310(10):609–613. doi: 10.1056/NEJM198403083101001. [DOI] [PubMed] [Google Scholar]
  35. Verstraete M., Bernard R., Bory M., Brower R. W., Collen D., de Bono D. P., Erbel R., Huhmann W., Lennane R. J., Lubsen J. Randomised trial of intravenous recombinant tissue-type plasminogen activator versus intravenous streptokinase in acute myocardial infarction. Report from the European Cooperative Study Group for Recombinant Tissue-type Plasminogen Activator. Lancet. 1985 Apr 13;1(8433):842–847. doi: 10.1016/s0140-6736(85)92208-1. [DOI] [PubMed] [Google Scholar]
  36. Weitz J. I., Koehn J. A., Canfield R. E., Landman S. L., Friedman R. Development of a radioimmunoassay for the fibrinogen-derived peptide B beta 1-42. Blood. 1986 Apr;67(4):1014–1022. [PubMed] [Google Scholar]
  37. Weitz J. I., Landman S. L., Crowley K. A., Birken S., Morgan F. J. Development of an assay for in vivo human neutrophil elastase activity. Increased elastase activity in patients with alpha 1-proteinase inhibitor deficiency. J Clin Invest. 1986 Jul;78(1):155–162. doi: 10.1172/JCI112545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Williams D. O., Borer J., Braunwald E., Chesebro J. H., Cohen L. S., Dalen J., Dodge H. T., Francis C. K., Knatterud G., Ludbrook P. Intravenous recombinant tissue-type plasminogen activator in patients with acute myocardial infarction: a report from the NHLBI thrombolysis in myocardial infarction trial. Circulation. 1986 Feb;73(2):338–346. doi: 10.1161/01.cir.73.2.338. [DOI] [PubMed] [Google Scholar]
  39. Wilner G. D., Nossel H. L., Canfield R. E., Butler V. P., Jr Immunochemical studies of human fibrinopeptide A using synthetic peptide homologues. Biochemistry. 1976 Mar 23;15(6):1209–1213. doi: 10.1021/bi00651a005. [DOI] [PubMed] [Google Scholar]
  40. Yudelman I., Greenberg J. Factors affecting fibrinopeptide-A levels in patients with venous thromboembolism during anticoagulant therapy. Blood. 1982 Apr;59(4):787–792. [PubMed] [Google Scholar]

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