Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1986 Oct 15;239(2):477–479. doi: 10.1042/bj2390477

An active-site titrant for human tissue-type plasminogen activator.

R A Smith
PMCID: PMC1147305  PMID: 3101668

Abstract

The reaction of recombinant tissue-type plasminogen activator with the inverse substrate 4-amidino-2-nitrophenyl 4'-anisate results in the rapid release of the chromogen 4-amidino-2-nitrophenol and the accumulation of the relatively stable 4-anisoyl-enzyme. Spectrophotometric monitoring of the reaction enables the operational molarity of the enzyme to be determined.

Full text

PDF
477

Selected References

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

  1. Bender M. L., Kézdy F. J., Wedler F. C. Alpha-chymotrypsin: enzyme concentration and kinetics. J Chem Educ. 1967 Feb;44(2):84–88. doi: 10.1021/ed044p84. [DOI] [PubMed] [Google Scholar]
  2. Browne M. J., Dodd I., Carey J. E., Chapman C. G., Robinson J. H. Increased yield of human tissue-type plasminogen activator obtained by means of recombinant DNA technology. Thromb Haemost. 1985 Aug 30;54(2):422–424. [PubMed] [Google Scholar]
  3. Chase T., Jr, Shaw E. Comparison of the esterase activities of trypsin, plasmin, and thrombin on guanidinobenzoate esters. Titration of the enzymes. Biochemistry. 1969 May;8(5):2212–2224. doi: 10.1021/bi00833a063. [DOI] [PubMed] [Google Scholar]
  4. Chase T., Jr, Shaw E. p-Nitrophenyl-p'-guanidinobenzoate HCl: a new active site titrant for trypsin. Biochem Biophys Res Commun. 1967 Nov 30;29(4):508–514. doi: 10.1016/0006-291x(67)90513-x. [DOI] [PubMed] [Google Scholar]
  5. Collen D., Stassen J. M., Verstraete M. Thrombolysis with human extrinsic (tissue-type) plasminogen activator in rabbits with experimental jugular vein thrombosis. Effect of molecular form and dose of activator, age of the thrombus, and route of administration. J Clin Invest. 1983 Feb;71(2):368–376. doi: 10.1172/JCI110778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cook R. R., Powers J. C. Benzyl p-guanidinothiobenzoate hydrochloride, a new active-site titrant for trypsin and trypsin-like enzymes. Biochem J. 1983 Nov 1;215(2):287–294. doi: 10.1042/bj2150287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dupe R. J., English P. D., Smith R. A., Green J. Acyl-enzymes as thrombolytic agents in dog models of venous thrombosis and pulmonary embolism. Thromb Haemost. 1984 Apr 30;51(2):248–253. [PubMed] [Google Scholar]
  8. Dupe R. J., Green J., Smith R. A. Acylated derivatives of streptokinase-plasminogen activator complex as thrombolytic agents in a dog model of aged venous thrombosis. Thromb Haemost. 1985 Feb 18;53(1):56–59. [PubMed] [Google Scholar]
  9. Gaffney P. J., Curtis A. D. A collaborative study of a proposed international standard for tissue plasminogen activator (t-PA). Thromb Haemost. 1985 Feb 18;53(1):134–136. [PubMed] [Google Scholar]
  10. Granelli-Piperno A., Reich E. A study of proteases and protease-inhibitor complexes in biological fluids. J Exp Med. 1978 Jul 1;148(1):223–234. doi: 10.1084/jem.148.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hoylaerts M., Rijken D. C., Lijnen H. R., Collen D. Kinetics of the activation of plasminogen by human tissue plasminogen activator. Role of fibrin. J Biol Chem. 1982 Mar 25;257(6):2912–2919. [PubMed] [Google Scholar]
  12. Korninger C., Matsuo O., Suy R., Stassen J. M., Collen D. Thrombolysis with human extrinsic (tissue-type) plasminogen activator in dogs with femoral vein thrombosis. J Clin Invest. 1982 Mar;69(3):573–580. doi: 10.1172/JCI110483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Korninger C., Stassen J. M., Collen D. Turnover of human extrinsic (tissue-type) plasminogen activator in rabbits. Thromb Haemost. 1981 Oct;46(3):658–661. [PubMed] [Google Scholar]
  14. Matsuo O., Rijken D. C., Collen D. Thrombolysis by human tissue plasminogen activator and urokinase in rabbits with experimental pulmonary embolus. Nature. 1981 Jun 18;291(5816):590–591. doi: 10.1038/291590a0. [DOI] [PubMed] [Google Scholar]
  15. Pennica D., Holmes W. E., Kohr W. J., Harkins R. N., Vehar G. A., Ward C. A., Bennett W. F., Yelverton E., Seeburg P. H., Heyneker H. L. Cloning and expression of human tissue-type plasminogen activator cDNA in E. coli. Nature. 1983 Jan 20;301(5897):214–221. doi: 10.1038/301214a0. [DOI] [PubMed] [Google Scholar]
  16. Smith R. A., Dupe R. J., English P. D., Green J. Acyl-enzymes as thrombolytic agents in a rabbit model of venous thrombosis. Thromb Haemost. 1982 Jun 28;47(3):269–274. [PubMed] [Google Scholar]
  17. Smith R. A., Dupe R. J., English P. D., Green J. Fibrinolysis with acyl-enzymes: a new approach to thrombolytic therapy. Nature. 1981 Apr 9;290(5806):505–508. doi: 10.1038/290505a0. [DOI] [PubMed] [Google Scholar]
  18. Van de Werf F., Ludbrook P. A., Bergmann S. R., Tiefenbrunn A. J., Fox K. A., de Geest H., Verstraete M., Collen D., Sobel B. E. Coronary thrombolysis with tissue-type plasminogen activator in patients with evolving myocardial infarction. N Engl J Med. 1984 Mar 8;310(10):609–613. doi: 10.1056/NEJM198403083101001. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES