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. 1971 Oct;50(10):2084–2090. doi: 10.1172/JCI106702

Separation of Plasma Thromboplastin Antecedent from Kallikrein by the Plasma α2-Macroglobulin, Kallikrein Inhibitor

Peter C Harpel 1
PMCID: PMC292142  PMID: 5165598

Abstract

Plasma thromboplastin antecedent (PTA, factor XI) is an important intermediate in the intrinsic coagulation system, and plasma kallikrein has been implicated as a mediator of the inflammatory process. Whereas their biologic activities are functionally distinct, their identity as separate entities in plasma has not been fully established, and the nature of their plasma inhibitors has not been completely characterized. A partially purified preparation containing the clotting, tosyl arginine methyl ester (TAMe) esterase and kinin-producing activities of these substances has been prepared by DEAE-cellulose chromatography of a Celite eluate obtained from acid-treated human plasma. These activities were not separable by acrylamide gel electrophoresis nor by isoelectric focusing, their pI being approximately 8.7. Human plasma α2-macroglobulin has been shown to inhibit the proteolytic activity of kallikrein and to inhibit partially its TAMe esterase activity. An α2-macroglobulin, PTA, kallikrein incubation mixture was separated by gel filtration chromatography. The α2-macroglobulin formed a high molecular weight complex with kallikrein and appeared in early chromatographic fractions. The PTA-clotting activity was not inhibited by the α2-macroglobulin; 64% of the initial PTA activity was isolated in later fractions free of kallikrein-induced kinin-like activity. In contrast, clotting, TAMe esterase, and kinin-forming activities were inhibited after gel filtration chromatography of an incubation mixture of these activities and partially purified C1̄ inactivator (C1 esterase inhibitor). Electrofocusing of an incubation mixture of an activated PTA, kallikrein preparation, and α2-macroglobulin resulted in the isolation of a PTA fraction free of kallikrein proteolytic activity, and with 4% of the original TAMe esterase activity. In this manner, activated PTA and plasma kallikrein have been shown to be distinct substances, and methods have been introduced for the further purification of active coagulation factor XI.

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Selected References

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  1. ALKJAERSIG N., FLETCHER A. P., SHERRY S. xi-Aminocaproic acid: an inhibitor of plasminogen activation. J Biol Chem. 1959 Apr;234(4):832–837. [PubMed] [Google Scholar]
  2. BACHMANN F., DUCKERT F., KOLLER F. The Stuart-Prower factor assay and its clinical significance. Thromb Diath Haemorrh. 1958 May 1;2(1-2):24–38. [PubMed] [Google Scholar]
  3. Colman R. W., Mattler L., Sherry S. Studies on the prekallikrein (kallikreinogen)--kallikrein enzyme system of human plasma. I. Isolation and purification of plasma kallikreins. J Clin Invest. 1969 Jan;48(1):11–22. doi: 10.1172/JCI105959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Colman R. W., Mattler L., Sherry S. Studies on the prekallikrein (kallikreinogen)--kallikrein enzyme system of human plasma. II. Evidence relating the kaolin-activated arginine esterase to plasma kallikrein. J Clin Invest. 1969 Jan;48(1):23–32. doi: 10.1172/JCI105971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Forbes C. D., Pensky J., Ratnoff O. D. Inhibition of activated Hageman factor and activated plasma thromboplastin antecedent by purified serum C1 inactivator. J Lab Clin Med. 1970 Nov;76(5):809–815. [PubMed] [Google Scholar]
  6. Ganrot P. O. Inhibition of plasmin activity by alpha-2-macroglobulin. Clin Chim Acta. 1967 May;16(2):328–329. doi: 10.1016/0009-8981(67)90201-x. [DOI] [PubMed] [Google Scholar]
  7. Gigli I., Mason J. W., Colman R. W., Austen K. F. Interaction of plasma kallikrein with the C1 inhibitor. J Immunol. 1970 Mar;104(3):574–581. [PubMed] [Google Scholar]
  8. Harpel P. C. A sensitive colorimetric method for the measurement of serum C1 inactivator using the substrate N-alpha-acetyl-L-lysine methyl ester. J Immunol. 1970 Apr;104(4):1024–1030. [PubMed] [Google Scholar]
  9. Harpel P. C. C1 inactivator inhibition by plasmin. J Clin Invest. 1970 Mar;49(3):568–575. doi: 10.1172/JCI106267. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Harpel P. C. Human plasma alpha 2-macroglobulin. An inhibitor of plasma kallikrein. J Exp Med. 1970 Aug 1;132(2):329–352. doi: 10.1084/jem.132.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. KAGEN L. J., LEDDY J. P., BECKER E. L. THE PRESENCE OF TWO PERMEABILITY GLOBULINS IN HUMAN SERUM. J Clin Invest. 1963 Aug;42:1353–1361. doi: 10.1172/JCI104819. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. KAGEN L. J. SOME BIOCHEMICAL AND PHYSICAL PROPERTIES OF THE HUMAN PERMEABILITY GLOBULINS. Br J Exp Pathol. 1964 Dec;45:604–611. [PMC free article] [PubMed] [Google Scholar]
  13. KINGDON H. S., DAVIE E. W., RATNOFF O. D. THE REACTION BETWEEN ACTIVATED PLASMA THROMBOPLASTIN ANTECEDENT AND DIISOPROPYLPHOSPHOFLUORIDATE. Biochemistry. 1964 Feb;3:166–173. doi: 10.1021/bi00890a005. [DOI] [PubMed] [Google Scholar]
  14. Kaplan A. P., Austen K. F. A pre-albumin activator of prekallikrein. J Immunol. 1970 Oct;105(4):802–811. [PubMed] [Google Scholar]
  15. Kellermeyer R. W., Graham R. C., Jr Kinins--possible physiologic and pathologic roles in man. N Engl J Med. 1968 Oct 3;279(14):754–759. doi: 10.1056/NEJM196810032791406. [DOI] [PubMed] [Google Scholar]
  16. LANDERMAN N. S., WEBSTER M. E., BECKER E. L., RATCLIFFE H. E. Hereditary angioneurotic edema. II. Deficiency of inhibitor for serum globulin permeability factor and/or plasma kallikrein. J Allergy. 1962 Jul-Aug;33:330–341. doi: 10.1016/0021-8707(62)90032-1. [DOI] [PubMed] [Google Scholar]
  17. LEVY L. R., LEPOW I. H. Assay and properties of serum inhibitor of C'l-esterase. Proc Soc Exp Biol Med. 1959 Aug-Sep;101:608–611. doi: 10.3181/00379727-101-25034. [DOI] [PubMed] [Google Scholar]
  18. LEWIS J. H. SEPARATION AND MOLECULAR WEIGHT ESTIMATION OF COAGULATION AND FIBRINOLYTIC PROTEINS BY SEPHADEX GEL FILTRATION. Proc Soc Exp Biol Med. 1964 May;116:120–122. doi: 10.3181/00379727-116-29177. [DOI] [PubMed] [Google Scholar]
  19. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  20. Lanchantin G. F., Plesset M. L., Friedmann J. A., Hart D. W. Dissociation of esterolytic and clotting activities of thrombin by trypsin-binding macroglobulin. Proc Soc Exp Biol Med. 1966 Feb;121(2):444–449. doi: 10.3181/00379727-121-30800. [DOI] [PubMed] [Google Scholar]
  21. MARGOLIS J. Glass surface and blood coagulation. Nature. 1956 Oct 13;178(4537):805–806. doi: 10.1038/178805b0. [DOI] [PubMed] [Google Scholar]
  22. MARGOLIS J. The mode of action of Hageman factor in the release of plasma kinin. J Physiol. 1960 May;151:238–252. doi: 10.1113/jphysiol.1960.sp006434. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mackay M., Silk E., Mason B., Combridge B. S. Studies on protein fractions isolated from human plasma. Kininogenase activity in human plasma fractions isolated with ether and ethanol. Br J Haematol. 1970 Jan;18(1):29–44. doi: 10.1111/j.1365-2141.1970.tb01416.x. [DOI] [PubMed] [Google Scholar]
  24. McConnell D. J., Mason B. The isolation of human plasma prekallikrein. Br J Pharmacol. 1970 May;38(3):490–502. doi: 10.1111/j.1476-5381.1970.tb10591.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Nagasawa S., Takahashi H., Koida M., Suzuki T., Schoenmakers J. G. Partial purification of bovine plasma kallikreinogen, its activation by the Hageman factor. Biochem Biophys Res Commun. 1968 Aug 21;32(4):644–649. doi: 10.1016/0006-291x(68)90287-8. [DOI] [PubMed] [Google Scholar]
  26. OWREN P. A., AAS K. The control of dicumarol therapy and the quantitative determination of prothrombin and proconvertin. Scand J Clin Lab Invest. 1951;3(3):201–208. doi: 10.3109/00365515109060600. [DOI] [PubMed] [Google Scholar]
  27. Ogston D., Ogston C. M., Ratnoff O. D., Forbes C. D. Studies on a complex mechanism for the activation of plasminogen by kaolin and by chloroform: the participation of Hageman factor and additional cofactors. J Clin Invest. 1969 Oct;48(10):1786–1801. doi: 10.1172/JCI106145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. PROCTOR R. R., RAPAPORT S. I. The partial thromboplastin time with kaolin. A simple screening test for first stage plasma clotting factor deficiencies. Am J Clin Pathol. 1961 Sep;36:212–219. doi: 10.1093/ajcp/36.3.212. [DOI] [PubMed] [Google Scholar]
  29. RAPAPORT S. I., SCHIFFMAN S., PATCH M. J., WARE A. G. A simple, specific one-stage assay for plasma thromboplastin antecedent activity. J Lab Clin Med. 1961 May;57:771–780. [PubMed] [Google Scholar]
  30. RATNOFF O. D., DAVIE E. W., MALLETT D. L. Studies on the action of Hageman factor: evidence that activated Hageman factor in turn activates plasma thromboplastin antecedent. J Clin Invest. 1961 May;40:803–819. doi: 10.1172/JCI104314. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. RATNOFF O. D., DAVIE E. W. The purification of activated Hageman factor (activated factor XII). Biochemistry. 1962 Nov;1:967–975. doi: 10.1021/bi00912a005. [DOI] [PubMed] [Google Scholar]
  32. RATNOFF O. D., LEPOW I. H. Some properties of an esterase derived from preparations of the first component of complement. J Exp Med. 1957 Aug 1;106(2):327–343. doi: 10.1084/jem.106.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. RATNOFF O. D., MILES A. A. THE INDUCTION OF PERMEABILITY-INCREASING ACTIVITY IN HUMAN PLASMA BY ACTIVATED HAGEMAN FACTOR. Br J Exp Pathol. 1964 Jun;45:328–345. [PMC free article] [PubMed] [Google Scholar]
  34. REISFELD R. A., LEWIS U. J., WILLIAMS D. E. Disk electrophoresis of basic proteins and peptides on polyacrylamide gels. Nature. 1962 Jul 21;195:281–283. doi: 10.1038/195281a0. [DOI] [PubMed] [Google Scholar]
  35. Ratnoff O. D., Naff G. B. The conversion of C'IS to C'1 esterase by plasmin and trypsin. J Exp Med. 1967 Feb 1;125(2):337–358. doi: 10.1084/jem.125.2.337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Ratnoff O. D., Pensky J., Ogston D., Naff G. B. The inhibition of plasmin, plasma kallikrein, plasma permeability factor, and the C'1r subcomponent of the first component of complement by serum C'1 esterase inhibitor. J Exp Med. 1969 Feb 1;129(2):315–331. doi: 10.1084/jem.129.2.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Ratnoff O. D. The biology and pathology of the initial stages of blood coagulation. Prog Hematol. 1966;5:204–245. [PubMed] [Google Scholar]
  38. SCHEIDEGGER J. J. Une micro-méthode de l'immuno-electrophorèse. Int Arch Allergy Appl Immunol. 1955;7(2):103–110. [PubMed] [Google Scholar]
  39. SCHIFFMAN S., RAPAPORT S. I., WARE A. G., MEHL J. W. Separation of plasma thromboplastin antecedent (PTA) and Hageman factor (HF) from human plasma. Proc Soc Exp Biol Med. 1960 Dec;105:453–455. doi: 10.3181/00379727-105-26138. [DOI] [PubMed] [Google Scholar]
  40. Vesterberg O., Svensson H. Isoelectric fractionation, analysis, and characterization of ampholytes in natural pH gradients. IV. Further studies on the resolving power in connection with separation of myoglobins. Acta Chem Scand. 1966;20(3):820–834. doi: 10.3891/acta.chem.scand.20-0820. [DOI] [PubMed] [Google Scholar]
  41. WEBSTER M. E., PIERCE J. V. Action of the kallikreins on synthetic ester substrates. Proc Soc Exp Biol Med. 1961 May;107:186–191. doi: 10.3181/00379727-107-26575. [DOI] [PubMed] [Google Scholar]
  42. Webster M. E. Human plasma kallikrein, its activation and pathological role. Fed Proc. 1968 Jan-Feb;27(1):84–89. [PubMed] [Google Scholar]

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