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. 1979 Dec 1;183(3):555–559. doi: 10.1042/bj1830555

A new fluorogenic substrate for plasmin.

P A Pierzchala, C P Dorn, M Zimmerman
PMCID: PMC1161636  PMID: 161507

Abstract

A new fluorogenic peptide substrate for plasmin, 7-(N-succinoylalanylphenylalanyl-lysylamido)-4-methylcoumarin trifluoroacetate salt, was prepared that can be used in a simple and direct assay. The results obtained by the assay method are linear over a wide range of enzyme concentrations and sensitive enough to detect as little as 10(-5) CTA units of plasmin. By making use of the inhibitor Trasylol and the differences in kinetic constants, plasmin can be specifically assayed even in the presence of the plasminogen activator thrombin, as well as in culture fluids from HeLa cells.

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Selected References

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

  1. ASTRUP T., MULLERTZ S. The fibrin plate method for estimating fibrinolytic activity. Arch Biochem Biophys. 1952 Oct;40(2):346–351. doi: 10.1016/0003-9861(52)90121-5. [DOI] [PubMed] [Google Scholar]
  2. Bell P. H., Dziobkowski C. T., Englert M. E. A sensitive fluorometric assay for plasminogen, plasmin, and streptokinase. Anal Biochem. 1974 Sep;61(1):200–208. doi: 10.1016/0003-2697(74)90346-7. [DOI] [PubMed] [Google Scholar]
  3. Brenner B. M. Functional and structural determinants of glomerular filtration. A brief historical perspective. Fed Proc. 1977 Nov;36(12):2599–2601. [PubMed] [Google Scholar]
  4. Clavin S. A., Bobbitt J. L., Shuman R. T., Smithwick E. L., Jr Use of peptidyl-4-methoxy-2-naphthylamides to assay plasmin. Anal Biochem. 1977 Jun;80(2):355–365. doi: 10.1016/0003-2697(77)90656-x. [DOI] [PubMed] [Google Scholar]
  5. Feinstein G., Kupfer A., Sokolovsky M. N-acetyl-(L-Ala) 3 -p-nitroanilide as a new chromogenic substrate for elastase. Biochem Biophys Res Commun. 1973 Feb 20;50(4):1020–1026. doi: 10.1016/0006-291x(73)91508-8. [DOI] [PubMed] [Google Scholar]
  6. Johnson A. J., Kline D. L., Alkjaersig N. Assay methods and standard preparations for plasmin, plasminogen and urokinase in purified systems, 1967-1968. Thromb Diath Haemorrh. 1969 Apr 30;21(2):259–272. [PubMed] [Google Scholar]
  7. Mattler L. E., Bang N. U. Serine protease specificity for peptide chromogenic substrates. Thromb Haemost. 1977 Dec 15;38(4):776–792. [PubMed] [Google Scholar]
  8. Nieuwenhuizen W., Wijngaards G., Groeneveld E. Flourogenic peptide amide substrates for the estimation of plasminogen activators and plasmin. Anal Biochem. 1977 Nov;83(1):143–148. doi: 10.1016/0003-2697(77)90519-x. [DOI] [PubMed] [Google Scholar]
  9. Robbins K. C., Summaria L. Plasminogen and plasmin. Methods Enzymol. 1976;45:257–273. doi: 10.1016/s0076-6879(76)45025-5. [DOI] [PubMed] [Google Scholar]
  10. Sherry S., Alkjaersig N., Fletcher A. P. [Activity of plasmin and streptokinase-activator on substituted arginine and lysine esters]. Thromb Diath Haemorrh. 1966 Jul 31;16(1):18–31. [PubMed] [Google Scholar]
  11. Stroud R. M. A family of protein-cutting proteins. Sci Am. 1974 Jul;231(1):74–88. doi: 10.1038/scientificamerican0774-74. [DOI] [PubMed] [Google Scholar]
  12. Zimmerman M., Quigley J. P., Ashe B., Dorn C., Goldfarb R., Troll W. Direct fluorescent assay of urokinase and plasminogen activators of normal and malignant cells: kinetics and inhibitor profiles. Proc Natl Acad Sci U S A. 1978 Feb;75(2):750–753. doi: 10.1073/pnas.75.2.750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Zimmerman M., Yurewicz E., Patel G. A new fluorogenic substrate for chymotrypsin. Anal Biochem. 1976 Jan;70(1):258–262. doi: 10.1016/s0003-2697(76)80066-8. [DOI] [PubMed] [Google Scholar]

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