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. 1982 Aug;70(2):412–423. doi: 10.1172/JCI110631

Catabolic Pathways for Streptokinase, Plasmin, and Streptokinase Activator Complex in Mice

IN VIVO REACTION OF PLASMINOGEN ACTIVATOR WITH α2-MACROGLOBULIN

Steven L Gonias 1,2,3, Monica Einarsson 1,2,3, Salvatore V Pizzo 1,2,3
PMCID: PMC371250  PMID: 6178757

Abstract

The catabolic pathways of streptokinase, plasmin, and activator complex prepared with human plasminogen were studied in mice. 125I-streptokinase clearance occurred in the liver and was 50% complete in 15 min. Incubation with mouse plasma had no effect on the streptokinase clearance rate. Complexes of plasmin and α2-plasmin inhibitor were eliminated from the plasma by a specific and saturable pathway. Competition experiments demonstrated that this pathway is responsible for the clearance of injected plasmin. Streptokinase-plasminogen activator complex formed with either 125I-plasminogen or 125I-streptokinase cleared in the liver at a significantly faster rate than either of the uncomplexed proteins (50% clearance in <3 min). Streptokinase incubated with human plasma also demonstrated this accelerated clearance. p-Nitrophenyl-p′-guanidinobenzoate-HCl or pancreatic trypsin inhibitor-treated complex cleared slowly compared with untreated complex independent of which protein was radiolabeled. Significant competition for clearance was demonstrated between α2-macroglobulin-trypsin and activator complex only when the plasmin(ogen) was the radiolabeled moiety. Large molar excesses of α2-plasmin inhibitor-plasmin failed to retard the clearance of activator complex. Hepatic binding of streptokinase-plasmin, in liver perfusion experiments, was dependent upon prior incubation with plasma (8-10% uptake compared to a background of ∼ 2.5%). Substitution of human α2-macroglobulin for plasma also resulted in binding when the incubation was performed for 10 min at 37°C (7.5%). Electrophoresis experiments confirmed the transfer of 0.8 mol plasmin/mol α2-macroglobulin when activator complex was incubated at 37°C with α2-macroglobulin for 40 min. Streptokinase transfer from activator complex to α2-macroglobulin was negligible. The in vivo clearance of activator complex is proposed to involve active attack of the complex on the α2-macroglobulin “bait region,” resulting in facilitated plasmin transfer. Dissociated streptokinase is rapidly bound and cleared by sites in the liver.

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Selected References

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  1. Aoki N., Moroi M., Tachiya K. Effects of alpha2-plasmin inhibitor on fibrin clot lysis. Its comparison with alpha2-macroglobulin. Thromb Haemost. 1978 Feb 28;39(1):22–31. [PubMed] [Google Scholar]
  2. BACK N., AMBRUS J. L., MINK I. B. Distribution and fate of I-131-labeled components of the fibrinolysin system. Circ Res. 1961 Nov;9:1208–1216. doi: 10.1161/01.res.9.6.1208. [DOI] [PubMed] [Google Scholar]
  3. Bajaj A. P., Castellino F. J. Activation of human plasminogen by equimolar levels of streptokinase. J Biol Chem. 1977 Jan 25;252(2):492–498. [PubMed] [Google Scholar]
  4. Barrett A. J., Brown M. A., Sayers C. A. The electrophoretically 'slow' and 'fast' forms of the alpha 2-macroglobulin molecule. Biochem J. 1979 Aug 1;181(2):401–418. doi: 10.1042/bj1810401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barrett A. J., Starkey P. M. The interaction of alpha 2-macroglobulin with proteinases. Characteristics and specificity of the reaction, and a hypothesis concerning its molecular mechanism. Biochem J. 1973 Aug;133(4):709–724. doi: 10.1042/bj1330709. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brockway W. J., Castellino F. J. Measurement of the binding of antifibrinolytic amino acids to various plasminogens. Arch Biochem Biophys. 1972 Jul;151(1):194–199. doi: 10.1016/0003-9861(72)90488-2. [DOI] [PubMed] [Google Scholar]
  7. Cederholm-Williams S. A., De Cock F., Lijnen H. R., Collen D. Kinetics of the reactions between streptokinase, plasmin and alpha 2-antiplasmin. Eur J Biochem. 1979 Oct;100(1):125–132. doi: 10.1111/j.1432-1033.1979.tb02040.x. [DOI] [PubMed] [Google Scholar]
  8. Chase T., Jr, Shaw E. p-Nitrophenyl-p'-guanidinobenzoate HCl: a new active site titrant for trypsin. Biochem Biophys Res Commun. 1967 Nov 30;29(4):508–514. doi: 10.1016/0006-291x(67)90513-x. [DOI] [PubMed] [Google Scholar]
  9. Coates G., DeNardo S. J., DeNardo G. L., Troy F. A. Pharmacokinetics of radioiodinated streptokinase. J Nucl Med. 1975 Feb;16(2):136–142. [PubMed] [Google Scholar]
  10. David G. S., Reisfeld R. A. Protein iodination with solid state lactoperoxidase. Biochemistry. 1974 Feb 26;13(5):1014–1021. doi: 10.1021/bi00702a028. [DOI] [PubMed] [Google Scholar]
  11. De Renzo E. C., Siiteri P. K., Hutchings B. L., Bell P. H. Preparation and certain properties of highly purified streptokinase. J Biol Chem. 1967 Feb 10;242(3):533–542. [PubMed] [Google Scholar]
  12. Deutsch D. G., Mertz E. T. Plasminogen: purification from human plasma by affinity chromatography. Science. 1970 Dec 4;170(3962):1095–1096. doi: 10.1126/science.170.3962.1095. [DOI] [PubMed] [Google Scholar]
  13. Einarsson M., Skoog B., Forsberg B., Einarsson R. Characterization of highly purified native streptokinase and altered streptokinase after alkaline treatment. Biochim Biophys Acta. 1979 May 10;568(1):19–29. doi: 10.1016/0005-2744(79)90269-9. [DOI] [PubMed] [Google Scholar]
  14. FLETCHER A. P., ALKJAERSIG N., SMYRNIOTIS F. E., SHERRY S. The treatment of patients suffering from early myocardial infarction with massive and prolonged streptokinase therapy. Trans Assoc Am Physicians. 1958;71:287–296. [PubMed] [Google Scholar]
  15. Fuchs H. E., Shifman M. A., Pizzo S. V. In vivo catabolism of alpha 1-proteinase inhibitor-trypsin, antithrombin III-thrombin and alpha 2-macroglobulin-methylamine. Biochim Biophys Acta. 1982 May 27;716(2):151–157. doi: 10.1016/0304-4165(82)90263-x. [DOI] [PubMed] [Google Scholar]
  16. Gonias S. L., Pizzo S. V. Altered clearance of human alpha 2-macroglobulin complexes following reaction with cis-dichlorodiamineplatinum(II). Biochim Biophys Acta. 1981 Dec 4;678(2):268–274. doi: 10.1016/0304-4165(81)90216-6. [DOI] [PubMed] [Google Scholar]
  17. Gonzalez-Gronow M., Siefring G. E., Jr, Castellino F. J. Mechanism of activation of human plasminogen by the activator complex, streptokinase-plasmin. J Biol Chem. 1978 Feb 25;253(4):1090–1094. [PubMed] [Google Scholar]
  18. Hall P. K., Roberts R. C. Physical and chemical properties of human plasma alpha2-macroglobulin. Biochem J. 1978 Jul 1;173(1):27–38. doi: 10.1042/bj1730027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Harpel P. C. Alpha2-plasmin inhibitor and alpha2-macroglobulin-plasmin complexes in plasma. Quantitation by an enzyme-linked differential antibody immunosorbent assay. J Clin Invest. 1981 Jul;68(1):46–55. doi: 10.1172/JCI110253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Harpel P. C., Hayes M. B., Hugli T. E. Heat-induced fragmentation of human alpha 2-macroglobulin. J Biol Chem. 1979 Sep 10;254(17):8669–8678. [PubMed] [Google Scholar]
  21. Harpel P. C. Plasmin inhibitor interactions. The effectiveness of alpha2-plasmin inhibitor in the presence of alpha2-macroglobulin. J Exp Med. 1977 Oct 1;146(4):1033–1040. doi: 10.1084/jem.146.4.1033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Harpel P. C. Studies on human plasma alpha 2-macroglobulin-enzyme interactions. Evidence for proteolytic modification of the subunit chain structure. J Exp Med. 1973 Sep 1;138(3):508–521. doi: 10.1084/jem.138.3.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. IIO M., WAGNER H. N., Jr, SCHEFFEL U., JABBOUR B. Studies of the reticuloendothelial system (RES). I. Measurement of the phagocytic capacity of the RES in man and dog. J Clin Invest. 1963 Mar;42:417–426. doi: 10.1172/JCI104729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Imber M. J., Pizzo S. V. Clearance and binding of two electrophoretic "fast" forms of human alpha 2-macroglobulin. J Biol Chem. 1981 Aug 10;256(15):8134–8139. [PubMed] [Google Scholar]
  25. JOHNSON A. J., McCARTY W. R. The lysis of artificially induced intravascular clots in man by intravenous infusions of streptokinase. J Clin Invest. 1959 Sep;38:1627–1643. doi: 10.1172/JCI103941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kaplan J., Nielsen M. L. Analysis of macrophage surface receptors. I. Binding of alpha-macroglobulin . protease complexes to rabbit alveolar macrophages. J Biol Chem. 1979 Aug 10;254(15):7323–7328. [PubMed] [Google Scholar]
  27. Lollar P., Owen W. G. Clearance of thrombin from circulation in rabbits by high-affinity binding sites on endothelium. Possible role in the inactivation of thrombin by antithrombin III. J Clin Invest. 1980 Dec;66(6):1222–1230. doi: 10.1172/JCI109973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McClintock D. K., Bell P. H. The mechanism of activation of human plasminogen by streptokinase. Biochem Biophys Res Commun. 1971 May 7;43(3):694–702. doi: 10.1016/0006-291x(71)90670-x. [DOI] [PubMed] [Google Scholar]
  29. Moroi M., Aoki N. Isolation and characterization of alpha2-plasmin inhibitor from human plasma. A novel proteinase inhibitor which inhibits activator-induced clot lysis. J Biol Chem. 1976 Oct 10;251(19):5956–5965. [PubMed] [Google Scholar]
  30. Müllertz S., Clemmensen I. The primary inhibitor of plasmin in human plasma. Biochem J. 1976 Dec 1;159(3):545–553. doi: 10.1042/bj1590545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nelles L. P., Hall P. K., Roberts R. C. Human alpha-2-macroglobulin. Studies on the electrophoretic heterogeneity. Biochim Biophys Acta. 1980 May 29;623(1):46–56. doi: 10.1016/0005-2795(80)90006-9. [DOI] [PubMed] [Google Scholar]
  32. Ohlsson K. Elimination of 125-I-trypsin alpha-macroglobulin complexes from blood by reticuloendothelial cells in dogs. Acta Physiol Scand. 1971 Feb;81(2):269–272. doi: 10.1111/j.1748-1716.1971.tb04900.x. [DOI] [PubMed] [Google Scholar]
  33. Reddy K. N., Markus G. Mechanism of activation of human plasminogen by streptokinase. Presence of active center in streptokinase-plasminogen complex. J Biol Chem. 1972 Mar 25;247(6):1683–1691. [PubMed] [Google Scholar]
  34. Renston R. H., Jones A. L., Christiansen W. D., Hradek G. T., Underdown B. J. Evidence for a vesicular transport mechanism in hepatocytes for biliary secretion of immunoglobulin A. Science. 1980 Jun 13;208(4449):1276–1278. doi: 10.1126/science.7375938. [DOI] [PubMed] [Google Scholar]
  35. Salvesen G. S., Barrett A. J. Covalent binding of proteinases in their reaction with alpha 2-macroglobulin. Biochem J. 1980 Jun 1;187(3):695–701. doi: 10.1042/bj1870695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Shifman M. A., Pizzo S. V. The in vivo metabolism of antithrombin III and antithrombin III complexes. J Biol Chem. 1982 Mar 25;257(6):3243–3248. [PubMed] [Google Scholar]
  37. Sjöholm I. Studies on the conformational changes of plasminogen induced during activation to plasmin and by 6-aminohexanoic acid. Eur J Biochem. 1973 Nov 15;39(2):471–479. doi: 10.1111/j.1432-1033.1973.tb03146.x. [DOI] [PubMed] [Google Scholar]
  38. Som P., Rhodes B. A., Bell W. R. Radiolabeled streptokinase and urokinase and their comparative biodistribution. Thromb Res. 1975 Mar;6(3):247–253. doi: 10.1016/0049-3848(75)90072-9. [DOI] [PubMed] [Google Scholar]
  39. Sottrup-Jensen L., Petersen T. E., Magnusson S. Mechanism of proteinase complex formation with alpha 2-macroglobulin. Three modes of trypsin binding. FEBS Lett. 1981 Jun 1;128(1):127–132. doi: 10.1016/0014-5793(81)81097-6. [DOI] [PubMed] [Google Scholar]
  40. Summaria L., Arzadon L., Bernabe P., Robbins K. C. The interaction of streptokinase with human, cat, dog, and rabbit plasminogens. The fragmentation of streptokinase in the equimolar plasminogen-streptokinase complexes. J Biol Chem. 1974 Aug 10;249(15):4760–4769. [PubMed] [Google Scholar]
  41. Tillett W. S., Sherry S. THE EFFECT IN PATIENTS OF STREPTOCOCCAL FIBRINOLYSIN (STREPTOKINASE) AND STREPTOCOCCAL DESOXYRIBONUCLEASE ON FIBRINOUS, PURULENT, AND SANGUINOUS PLEURAL EXUDATIONS. J Clin Invest. 1949 Jan;28(1):173–190. doi: 10.1172/JCI102046. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Violand B. N., Castellino F. J. Mechanism of the urokinase-catalyzed activation of human plasminogen. J Biol Chem. 1976 Jul 10;251(13):3906–3912. [PubMed] [Google Scholar]
  43. Wiman B., Collen D. On the kinetics of the reaction between human antiplasmin and plasmin. Eur J Biochem. 1978 Mar 15;84(2):573–578. doi: 10.1111/j.1432-1033.1978.tb12200.x. [DOI] [PubMed] [Google Scholar]
  44. Wiman B., Collen D. On the mechanism of the reaction between human alpha 2-antiplasmin and plasmin. J Biol Chem. 1979 Sep 25;254(18):9291–9297. [PubMed] [Google Scholar]
  45. Wiman B. On the reaction of plasmin or plasmin-streptokinase complex with aprotinin or alpha 2-antiplasmin. Thromb Res. 1980 Jan 1;17(1-2):143–152. doi: 10.1016/0049-3848(80)90302-3. [DOI] [PubMed] [Google Scholar]
  46. Wulf R. J., Mertz E. T. Studies on plasminogen. 8. Species specificity of streptokinase. Can J Biochem. 1969 Oct;47(10):927–931. doi: 10.1139/o69-145. [DOI] [PubMed] [Google Scholar]
  47. Wyckoff M., Rodbard D., Chrambach A. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate-containing buffers using multiphasic buffer systems: properties of the stack, valid Rf- measurement, and optimized procedure. Anal Biochem. 1977 Apr;78(2):459–482. doi: 10.1016/0003-2697(77)90107-5. [DOI] [PubMed] [Google Scholar]

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