Kinins IV pp 283-289 | Cite as

Individual Reaction Steps in the Release of Kallidin from Kininogen by Tissue Kallikrein

  • Franz Fiedler
  • Heide Hinz
  • Friedrich Lottspeich
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 198A)


At low pH values (around 6), porcine pancreatic β-kallikrein B attacks at first the C-terminal ARg bond of the kinin moiety in bovine HMW kininogen. Arg-cleaved kininogen accumulates as an intermediate in the solution. Kallidin is released by cleavage of the aminoterminal Met-Lys bond in a second step. At pH values between 7.6 and 9, however, Arg-cleaved kininogen does not occur as a free intermediate. The participation as a (free, not only enzyme-bound) intermediate of Arg-cleaved kininogen in a short-lived especially reactive conformation or of Met-cleaved kininogen is also unlikely. Probably, both the Met and the Arg bonds are hydrolyzed in one enzyme-substrate complex which does not dissociate between these two events.

Kinetic constants for the release of kallidin from native single-chain HMW kininogen and from Arg-cleaved kininogen (even if this Arg residue is removed) remarkably have the same values. Evidently, the rate of the reaction is determined by steps leading to the hydrolysis of the Met bond. As the state of the C-terminal Arg residue has no influence, the efficient cleavage of the Met bond by tissue kallirkein is probably not due to some strain in the kininogen molecule in the region of this bond. As modification of Arg residues of kininogen prevents cleavage also of the Met bond, some Arg residue(s) appear(s) to play a crucial role in this process. Kcat/Km (1.4 x 106 M-1 sec-1 at pH 9, 25° C) is very high for a proteolytic reaction, mainly because of the low value of (0.6 μM). Kcat/Km for the hydrolysis of the Met bond in kininogen is 3 500 times higher than in the peptide Ser-Leu-Met-Lys-brady-kinin with a partial kininogen sequence. Important interactions with the enzyme thus occur in regions of the kininogen molecule outside this sequence. Kcat/Km for the hydrolysis of the Arg bond in the kininogen peptide Pro-Phe-Arg-Ser-Val-Gln is also 14 times lower than this constant for the release of kallidin from kininogen, but kcat is 6 times higher. This can explain why cleavage of the Arg bond is not reflected in the kinetic constants of single-chain kininogen.


Kinetic Constant Tissue Kallikrein High Molecular Weight Kininogen Proteolytic Reaction Free Intermediate 
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  1. 1.
    E. Habermann, Strukturaufklarung kininliefernder Peptide aus Rinderserum-Kininogen, Arch. Exp. Path. Pharmak., 253: 474–483 (1966).Google Scholar
  2. 2.
    Y. N. Han, H. Kato, S. Iwanaga, and T. Suzuki, Bovine plasma high molecular weight kininogen: The amino acid sequence of fragment 1 (glycopeptide) released by the action of plasma kallikrein and its location in the precursor protein, FEBS Lett., 63: 197 – 200 (1976).PubMedCrossRefGoogle Scholar
  3. 3.
    F. Fiedler and G. Leysath, Substrate specificity of porcine pancreatic kallikrein, Adv. Exp. Med. Biol., 120A: 261–271 (1979).Google Scholar
  4. 4.
    F. Fiedler, Enzymology of porcine tissue kallikrein, Adv. Exp. Med. Biol., 156A: 263 – 274 (1983).PubMedGoogle Scholar
  5. 5.
    F. Fiedler, E. Fink, H. Tschesche, and H. Fritz, Porcine glandular kallikreins, Methods Enzymol., 80: 493 – 532 (1981).PubMedCrossRefGoogle Scholar
  6. 6.
    C. Kutzbach and G. Schmidt-Kastner, Kallikrein from pig pancreas: Purification, separation of components A and B, and crystallization, Hoppe-Seyler1s Z. Physiol. Chem., 353: 1099 – 1106 (1972).CrossRefGoogle Scholar
  7. 7.
    M. Komiya, H. Kato, and T. Suzuki, Bovine plasma kininogens. I. Further purification of high molecular weight kininogen and its physiocochemical properties, J. Biochem. (Tokyo), 76: 811 – 822 (1974).Google Scholar
  8. 8.
    T. Shimada, T. Sugo, H. Kato, and S. Iwanaga, A method for preparation of a single chain high-molecular-weight (HMW) kininogen from bovine plasma, J. Biochem. (Tokyo), 92: 679 - 688 (1982).Google Scholar
  9. 9.
    K. Weber and M. Osborn, The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis, J. Biol. Chem., 244: 4406–4412 (1969).PubMedGoogle Scholar
  10. 10.
    W. Miiller-Esterl, G. Rauth, H. Fritz, F. Lottspeich, and A. Henschen, Human kininogens, in: “Kininogenases-Kallikrein 6, ” G. L. Haberland, J. W. Rohen, H. Fritz, and P. Huber, eds., F. K. Schattauer Verlag (1983).Google Scholar
  11. 11.
    K. Mori, W. Sakamoto, and S. Nagasawa, Studies on human high molecular eight (HMW) kininogen. III. Cleavage of HMW kininogen by the action of human salivary kallikrein, J. Biochem. (Tokyo), 90: 503–509 (1981).Google Scholar
  12. 12.
    E. S. Prado, C. A. M. Sampaio, M. S. Araujo-Viel, and R. C. R. Stella, Characterization of horse urinary kallikrein, in: “Recent Progress on Kinins (Agents and Actions Supplements 9),” H. Fritz, G. Dietze, F. Fiedler, and G. L. Haberland, eds., Birkhauser Verlag (1982).Google Scholar
  13. 13.
    J. V. Pierce and J. A. Guimaraes, Further characterization of highly purified human plasma kininogens, in: “Chemistry and Biology of the Kallikrein-Kinin System in Health and Disease,” J. J. Pisano and K. F. Austen, eds., U. S. Government Printing Office, without year (1976).Google Scholar
  14. 14.
    M. Maier, K. F. Austen, and J. Spragg, Kinetic analysis of the interaction of human tissue kallikrein with single-chain human high and low molecular weight kininogens, Biochemistry, 80: 3928 – 3932 (1983).Google Scholar

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© Plenum Press, New York 1986

Authors and Affiliations

  • Franz Fiedler
    • 1
  • Heide Hinz
    • 1
  • Friedrich Lottspeich
    • 2
  1. 1.Abteilung für Klinische Chemie und Klinische Biochemie in der Chirurgischen Klinik InnenstadtUniversität MünchenMünchen 2Deutschland
  2. 2.Max-Planck-Institut für BiochemieMartinsriedDeutschland

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