TY - JOUR
T1 - Restoration of serine protease-inhibitor interaction by protein engineering
AU - Madison, Edwin L.
AU - Goldsmith, Elizabeth J.
AU - Gething, Mary Jane H
AU - Sambrook, Joseph F.
AU - Gerard, Robert D.
N1 - Copyright:
Copyright 2007 Elsevier B.V., All rights reserved.
PY - 1990/12/15
Y1 - 1990/12/15
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=0025597970&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0025597970&partnerID=8YFLogxK
M3 - Article
C2 - 2123870
AN - SCOPUS:0025597970
SN - 0021-9258
VL - 265
SP - 21423
EP - 21426
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 35
ER -