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. 1974 Dec;54(6):1444–1454. doi: 10.1172/JCI107892

Immunochemical Studies of Plasma Kallikrein

Andranik Bagdasarian 1,2, Biswajit Lahiri 1,2, Richard C Talamo 1,2, Pat Wong 1,2, Robert W Colman 1,2
PMCID: PMC301700  PMID: 4140197

Abstract

A monospecific antibody against human plasma kallikrein has been prepared in rabbits with kallikrein further purified to remove gamma globulins. The antisera produced contained antikallikrein and also anti-IgG, in spite of only 8% contamination of kallikrein preparation with IgG. The latter antibody was removed by adsorption of antisera with either Fletcher factor-deficient plasma or with purified IgG. Both kallikrein and prekallikrein (in plasma) cross-react with the antibody with no apparent difference between the precipitation arcs developed during immunoelectrophoresis and no significant difference in reactivity when quantified by radial immunodiffusion.

Kallikrein antibody partially inhibits the esterolytic and fully inhibits the proteolytic activity of kallikrein. In addition, the antibody inhibits the activation of prekallikrein, as measured by esterase or kinin release. The magnitude of the inhibition is related to the molecular weight of the activator used. Thus, for the four activators tested, the greatest inhibition is observed with kaolin and factor XIIA, while large activator and the low molecular weight prekallikrein activators are less inhibited.

With the kallikrein antibody, the incubation of kallikrein with either plasma or partially purified C1 esterase inactivator results in a new precipitin arc, as detected by immunoelectrophoresis. This finding provides physical evidence for the interaction of the enzyme and inhibitor. No new arc could be demonstrated between kallikrein and α2-macroglobulin, or α1-antitrypsin, although the concentration of free kallikrein antigen decreases after interaction with the former inhibitor.

By radial immunodiffusion, plasma from healthy individuals contained 103±13 μg/ml prekallikrein antigen. Although in mild liver disease, functional and immunologic kallikrein are proportionally depressed, the levels of prekallikrein antigen in plasma samples from patients with severe liver disease remains 40% of normal, while the functional kallikrein activity was about 8%. These observations suggest that the livers of these patients have synthesized a proenzyme that cannot be converted to active kallikrein.

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Selected References

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

  1. Bagdasarian A., Lahiri B., Colman R. W. Origin of the high molecular weight activator of prekallikrein. J Biol Chem. 1973 Nov 25;248(22):7742–7747. [PubMed] [Google Scholar]
  2. Bagdasarian A., Talamo R. C., Colman R. W. Isolation of high molecular weight activators of human plasma prekallikrein. J Biol Chem. 1973 May 25;248(10):3456–3463. [PubMed] [Google Scholar]
  3. 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]
  4. COTRAN R. S., MAJNO G. A LIGHT AND ELECTRON MICROSCOPIC ANALYSIS OF VASCULAR INJURY. Ann N Y Acad Sci. 1964 Aug 27;116:750–764. doi: 10.1111/j.1749-6632.1964.tb52543.x. [DOI] [PubMed] [Google Scholar]
  5. Cochrane C. G., Wuepper K. D. The first component of the kinin-forming system in human and rabbit plasma. Its relationship to clotting factor XII (Hageman Factor). J Exp Med. 1971 Oct 1;134(4):986–1004. doi: 10.1084/jem.134.4.986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Colman R. W., Mason J. W., Sherry S. The kallikreinogen-kallikrein enzyme system of human plasma. Assay of components and observations in disease states. Ann Intern Med. 1969 Oct;71(4):763–773. doi: 10.7326/0003-4819-71-4-763. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Colman R. W., Mitchell B. Specificity of inhibitors lacking in alpha antitrypsin deficiency and hereditary angioedema toward human proteolytic enzymes. Clin Chim Acta. 1972 Jun;39(1):5–14. doi: 10.1016/0009-8981(72)90293-8. [DOI] [PubMed] [Google Scholar]
  10. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  11. 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]
  12. Groves W. E., Davis F. C., Jr, Sells B. H. Spectrophotometric determination of microgram quantities of protein without nucleic acid interference. Anal Biochem. 1968 Feb;22(2):195–210. doi: 10.1016/0003-2697(68)90307-2. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Harpel P. C. Separation of plasma thromboplastin antecedent from kallikrein by the plasma 2 -macroglobulin, kallikrein inhibitor. J Clin Invest. 1971 Oct;50(10):2084–2090. doi: 10.1172/JCI106702. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Kaplan A. P., Austen K. F. A pre-albumin activator of prekallikrein. J Immunol. 1970 Oct;105(4):802–811. [PubMed] [Google Scholar]
  17. Kaplan A. P., Kay A. B., Austen K. F. A prealbumin activator of prekallikrein. 3. Appearance of chemotactic activity for human neutrophils by the conversion of human prekallikrein to kallikrein. J Exp Med. 1972 Jan;135(1):81–97. doi: 10.1084/jem.135.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. 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]
  20. MEHL J. W., O'CONNELL W., DEGROOT J. MACROGLOBULIN FROM HUMAN PLASMA WHICH FORMS AN ENZYMATICALLY ACTIVE COMPOUND WITH TRYPSIN. Science. 1964 Aug 21;145(3634):821–822. doi: 10.1126/science.145.3634.821. [DOI] [PubMed] [Google Scholar]
  21. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  22. McConnell D. J. Inhibitors of kallikrein in human plasma. J Clin Invest. 1972 Jul;51(7):1611–1623. doi: 10.1172/JCI106962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. OUCHTERLONY O. Antigen-antibody reactions in gels. IV. Types of reactions in coordinated systems of diffusion. Acta Pathol Microbiol Scand. 1953;32(2):230–240. [PubMed] [Google Scholar]
  24. Ohlsson K. Interactions between bovine -chymotrypsin and the protease inhibitors of human and dog serum in vitro. Scand J Clin Lab Invest. 1971 Sep;28(1):5–11. doi: 10.3109/00365517109090656. [DOI] [PubMed] [Google Scholar]
  25. SCHEIDEGGER J. J. Une micro-méthode de l'immuno-electrophorèse. Int Arch Allergy Appl Immunol. 1955;7(2):103–110. [PubMed] [Google Scholar]
  26. SIEGELMAN A. M., CARLSON A. S., ROBERTSON T. Investigation of serum trypsin and related substances. I. The quantitative demonstration of trypsinlike activity in human blood serum by a micromethod. Arch Biochem Biophys. 1962 Apr;97:159–163. doi: 10.1016/0003-9861(62)90058-9. [DOI] [PubMed] [Google Scholar]
  27. Schreiber A. D., Kaplan A. P., Austen K. F. Plasma inhibitors of the components of the fibrinolytic pathway in man. J Clin Invest. 1973 Jun;52(6):1394–1401. doi: 10.1172/JCI107312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Talamo R. C., Haber E., Austen K. F. A radioimmunoassay for bradykinin in plasma and synovial fluid. J Lab Clin Med. 1969 Nov;74(5):816–827. [PubMed] [Google Scholar]
  29. Webster M. E., Gilmore J. P. The estimation of the kallidins in blood and urine. Biochem Pharmacol. 1965 Jul;14(7):1161–1163. doi: 10.1016/0006-2952(65)90046-8. [DOI] [PubMed] [Google Scholar]
  30. Wong P., Colman R. W., Talamo R. C., Babior B. M. Kallikrein-bradykinin system in chronic alcoholic liver disease. Ann Intern Med. 1972 Aug;77(2):205–209. doi: 10.7326/0003-4819-77-2-205. [DOI] [PubMed] [Google Scholar]

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