Kinins IV pp 299-308 | Cite as

New Synthetic Substrate for Kallikrein and its Application

  • Setsuro Fujii
  • Yuji Hitomi
  • Kenjiro Kimura
  • Masao Ishii
  • Masateru Kurumi
  • Takuo Aoyama
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 198A)

Summary

We developed a new synthetic substrate, Pro-Phe-Arg-α-naphthyl ester, for kallikrein. We found that this substrate had higher specificity and sensitivity for kallikrein and was applied for the preparation of zymogram and for the histochemical demonstration.

With Pro-Phe-Arg-α-NE as substrate, the minimum detectable concentration of human urinary kallikrein was about 0.001 KU and then kallikrein could be determined with 25 ul of human urine.

We also found the possibility that occurring of abnormalities during pregnancy were predicted by the determination of urinary kallikrein of pregnants.

Zymograms were prepared for various kinds of kallikrein using this substrate.

The localization of kallikrein-like enzyme in rat kidneys was defined by the application this substrate for histochemistry.

Moreover, cytochemical demonstrations of leucocytes in human blood were done using Ts-Lys-α-NE and Ac-Tyr-α-NE.

Keywords

Formalin Hydrolysis Glycerine Acetone Electrophoresis 

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References

  1. 1.
    R. M. Houseby and R. E. Smith, Synthetic oligopeptide substrates: Their diagnostic application in blood coagulation, fibrinolysis, and other pathologic state, Semi. Thrombo. Haemo., 6,175–314,(1980)CrossRefGoogle Scholar
  2. 2.
    Y. Hitomi, M. Niinobe, and S. Fujii, A sensitive colorimetric assay for human urinary kallikrein, Clin. Chim. Acta, 100, 275–283, (1980)PubMedCrossRefGoogle Scholar
  3. 3.
    M. Niinobe, Y. Hitomi, and S. Fujii, A sensitive colorimetric assay for various proteases using naphthyl ester derivatives as substrates, J. Biochem., 87, 779–783, (1980).PubMedGoogle Scholar
  4. 4.
    M. Saito, S. Munakata, I. Sada, Y. Nishimura, Y. Nakamura, Y. Nomura, and N. Shinagawa, Changes of urinary kallikrein levels during pregnancy, Sanfujinka no Sekai (in Japanese), 36, 173–176, (1983).Google Scholar
  5. 5.
    K. Kimura, M. Takagi, T. Igari, M. Ishii, T. Ikeda, T. Takeda, and S. Murano, Histochemical localization of kallikrein-like Pro-Phe- Arg-naphthyl ester esterase activity in the rat kidney, Histochem., 75, 91–98, (1982).CrossRefGoogle Scholar
  6. 6.
    T. Honda, Y. Hitomi, M. Niinobe, and S. Fujii, Cytochemical demon strations of protease in human peripheral blood cells by use of new a-naphthyl ester derivatives, Histochem., 77, 299–302, (1983).CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1986

Authors and Affiliations

  • Setsuro Fujii
    • 1
  • Yuji Hitomi
    • 1
  • Kenjiro Kimura
    • 2
  • Masao Ishii
    • 2
  • Masateru Kurumi
    • 3
  • Takuo Aoyama
    • 3
  1. 1.Division of Reguration of Macromolecular Function, Inst. for Protein Res.Osaka Univ.Suita, OsakaJapan
  2. 2.Second Dep. of Internal Med., Faculty of MedicineTokyo UniversityTokyoJapan
  3. 3.Research Labs.Torii & Co., Ltd.Ichikawa, ChibaJapan

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