Partial Purification of Rat Urinary Kininogenase and its Reactions with Active Center Reagents of Trypsin

  • Marcos Mares-Guia
  • Eder Silva
  • Carlos R. Diniz
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 8)


The kininogenase from rat urine has the properties of a trypsin-like enzyme, as determined with the help of synthetic substrates or competitive inhibitors (Mares-Guia and Diniz, 1967). The work done previously was carried out with a partially purified preparation of high specific activity. The purification procedure will now be described, as well as the behavior of the enzyme in the presence of specific reagents for the trypsin active center.


Methylene Blue Active Center Ethyl Ester Amidase Activity Chloromethyl Ketone 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Beraldo, W.T., R. L. Araujo, and M. Mares-Guia: Oxytocic esterase in urine. Am. J. Physiol. 211: 975–980 (1966).PubMedGoogle Scholar
  2. Chase, T. Jr., and E. Shaw: p-Nitrophenyl-p1 -guanidi-nobenzoate HCl: A new active site titrant for trypsin. Biochem. Biophys. Res. Commun. 29: 508–514 (1967).PubMedCrossRefGoogle Scholar
  3. Diniz, C. R. and I. F. Carvalho: A micromethod for determination of bradykininogen under several conditions. Ann. N. Y. Acad. Sci. 104: 77–89 (1963).PubMedCrossRefGoogle Scholar
  4. Diniz, C. R., A.A. Pereira, J. Barroso, and M. Mares-Guia: On the specificity of urinary kallikreins. Biochem. Biophys. Res. Commun. 21: 448–453 (1965).PubMedCrossRefGoogle Scholar
  5. Diniz, C.R., A.A. Pereira, J. Barroso, and M. Mares-Guia: Studies of the specificity of kinin-forming enzymes. The inhibition of the kininogenic activity of trypsin and kallikreins by model compounds, in Hypotensive Peptides, edited by E.G. Erdös, N. Back, and F. Sicuteri, Springer-Verlag, New York Inc., 175–183 (1966).CrossRefGoogle Scholar
  6. Fiedler, F., and E. Werle: Activation, inhibition, and pH-dependence of the hydrolysis of α -N-benzoyl-L-arginine ethyl ester catalyzed by kallikrein from procine pancreas. European J. Biochem. 7: 27–33 (1968).CrossRefGoogle Scholar
  7. Gutfreund, H.: The characterization of the catalytic site of trypsin. Trans. Faraday Soc. 51: 441–446 (1955).CrossRefGoogle Scholar
  8. Habermann, E.: Unterscheidung von Kallikrein und Kallikreinvorstuffe mittels Antiseren und Diisopropylfluorophosphats. Hoppe-Syler’s Z. Physiol. Chem. 328: 24–30 (1962).CrossRefGoogle Scholar
  9. Habermann, E.: Enzymatic kinin release from kininogen and from low-molecular compounds, in Hypotensive Peptides, edited by E. G. Erdbs, N. Back, and F. Sicuteri, Springer-Verlag New York Inc., 116–128 (1966).CrossRefGoogle Scholar
  10. Habermann, E. and G. Blennemann: Über substrate und Reaktion s produkte der kininbildenden Enzyme Trypsin, Serum- und Pankreaskallikrein sowie von crotalusgift. Naunyn-Schiederbergs Arch. Pharmakol. Exp. Pathol. 249: 357–373 (1964).CrossRefGoogle Scholar
  11. Habermann, E., und W. Klett: Reinigung und eigene Eigenschaften eines Kallikreins aus Schweine serum. Biochem. Z. 346: 133–158 (1966).PubMedGoogle Scholar
  12. Inagami, T., and J. M. Sturtevant: Nonspecific catalysis by α -chymotrypsin and trypsin. J. Biol. Chem. 235: 1019–1023, (1960).Google Scholar
  13. Ksohland, D. E. Jr., and K. E. Neet: The catalytic and regulatory properties of enzymes. Ann. Rev. Biochem. 37: 359–410 (1968).CrossRefGoogle Scholar
  14. Mares-Guia, M.: Hydrophobic interactions in the trypsin active center. The sensitivity of the hydrophobic binding site to side chain, modifications in competitive inhibitors of the Amidinium type. Arch. Biochem. Biophys. 127: 317–322 (1968).PubMedCrossRefGoogle Scholar
  15. Mares-Guia, M., and C. R. Diniz: Studies on the mechanism of rat urinary kallikrein catalysis, and its relation to catalysis by trypsin. Arch. Biochem. Biophys. 121: 750–756 (1967).PubMedCrossRefGoogle Scholar
  16. Mares-Guia, M., and E. Shaw: Studies on the active center of trypsin. The binding of amidines and guanidines as models of the substrate side chain. J. Biol. Chem. 240: 1579–1585 (1965).PubMedGoogle Scholar
  17. Mares-Guia, M., and E. Shaw: The specific inactivation of trypsin by ethyl p-guanidinobenzoate: J. Biol. Chem. 242: 5782–5788 (1967).PubMedGoogle Scholar
  18. Oosterbaan, R. A., and J. A. Cohen: The active stie of esterases, in Structure and Activity of Enzymes, edited by T. W. Goodwin, J.I. Harris, and B. S. Hartley, Academic Press, New York, 87–95 (1964).Google Scholar
  19. Pashkina, T. S., V. P. Zykova, T. P. Egorova, V. F. Nartikova, A. V. Karpova, and M. V. Guzeva: Reaction of N- α -tosyl-L-lysylchloromethane with kallikrein from human urine and saliva. Biokhimiya 33: 745–752 (1968).Google Scholar
  20. Pashkina, T. S., V. P. Zykova, T. P. Egorova, V. F. Nartikova, A. V. Karpova, and M. V. Guzeva: Reaction of N- α -tosyl-L-lysylchloromethane with kallikrein from human urine and saliva. See also Chem. Abs. 70: 420G, 1969.Google Scholar
  21. Pierce, J. V.: Structural features of plasma kinins and kininogens. Federation Proc. 27: 52–57 (1968).Google Scholar
  22. Prado, E. S., and J. L. Prado: Partial purification of a plasma kinin-forming enzyme from horse urine. Experientia 17: 1–3 (1961).CrossRefGoogle Scholar
  23. Shaw, E., M. Mares-Guia, and W. Cohen: Evidence for an active center histidine in trypsin through use of a specific reagent, TLCK, the chloromethyl ketone derived from N- α -tosyl-L-lysine. Biochemistry 4: 2219–2221 (1965).CrossRefGoogle Scholar
  24. Schoellman, G., and Shaw: Direct evidence for an active center histidine in the active center of chymotrypsin. Biochemistry 2: 252–255 (1963).CrossRefGoogle Scholar
  25. Werle, E., and W. Appel: Photochemische Inaktivierung von Kallikrein durch Methylenblau. Z. Vit. -, Hormon, Fermentforsch VIII, 24–28 (1956).Google Scholar

Copyright information

© Plenum Press, New York 1970

Authors and Affiliations

  • Marcos Mares-Guia
    • 1
  • Eder Silva
    • 1
  • Carlos R. Diniz
    • 1
  1. 1.Departamento de Bioquimica, Instituto de Ciencias BiologicasUniversidade Federal de Minas GeraisBelo HorizonteBrasil

Personalised recommendations