Restoration of serine protease-inhibitor interaction by protein engineering

Edwin L. Madison, Elizabeth J. Goldsmith, Mary Jane H Gething, Joseph F. Sambrook, Robert D. Gerard

Research output: Contribution to journalArticlepeer-review

79 Scopus citations


Tissue-type plasminogen activator (t-PA) catalyzes the rate-limiting step in the fibrinolytic cascade: conversion of plasminogen to plasmin. Plasma contains several inhibitors of t-PA that limit its activity and prevent systemic activation of plasminogen. The most important of these is endothelial cell plasminogen activator inhibitor (PAI-1), a member of the serine protease inhibitor (serpin) gene family. We have previously demonstrated that mutation of arginine 304 of t-PA to a glutamic acid residue drastically reduces the rate of interaction between the enzyme and its suicide substrate, PAI-1, without affecting the reactivity of the enzyme toward its normal substrate, plasminogen (Madison, E.L., Goldsmith, E.J., Gerard, R.D., Gething, M.J., and Sambrook, J.F. (1989) Nature 339, 721-724). We report here the use of protein modeling to design a compensatory mutation in PAI-1 (glutamic acid 350 to arginine) and create a molecule that rapidly inhibits this 'serpin-resistant', variant of t-PA.

Original languageEnglish (US)
Pages (from-to)21423-21426
Number of pages4
JournalJournal of Biological Chemistry
Issue number35
StatePublished - 1990

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology


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