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Science 10 February 1989:
Vol. 243. no. 4892, pp. 792 - 794
DOI: 10.1126/science.2916125
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Articles

Science, Vol 243, Issue 4892, 792-794
Copyright © 1989 by American Association for the Advancement of Science

articles

Control of enzyme activity by an engineered disulfide bond

M Matsumura and BW Matthews

Institute of Molecular Biology, University of Oregon, Eugene 97403.

A novel approach to the control of enzyme catalysis is presented in which a disulfide bond engineered into the active-site cleft of bacteriophage T4 lysozyme is capable of switching the activity on and off. Two cysteines (Thr21----Cys and Thr142----Cys) were introduced by oligonucleotide-directed mutagenesis into the active-site cleft. These cysteines spontaneously formed a disulfide bond under oxidative conditions in vitro, and the catalytic activity of the oxidized (cross-linked) T4 lysozyme was completely lost. On exposure to reducing agent, however, the disulfide bond was rapidly broken, and the reduced (non-cross-linked) lysozyme was restored to full activity. Thus an enzyme has been engineered such that redox potential can be used to control catalytic activity.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Use of experimental crystallographic phases to examine the hydration of polar and nonpolar cavities in T4 lysozyme.
L. Liu, M. L. Quillin, and B. W. Matthews (2008)
PNAS 105, 14406-14411
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Engineering the Refolding Pathway and the Quaternary Structure of Seminal Ribonuclease by Newly Introduced Disulfide Bridges.
A. Russo, A. Antignani, C. Giancola, and G. D'Alessio (2002)
J. Biol. Chem. 277, 48643-48649
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The Structural Bases of Antibiotic Resistance in the Clinically Derived Mutant beta -Lactamases TEM-30, TEM-32, and TEM-34.
X. Wang, G. Minasov, and B. K. Shoichet (2002)
J. Biol. Chem. 277, 32149-32156
   Abstract »    Full Text »    PDF »
Adjacent cysteine residues as a redox switch.
C. Park and R. T. Raines (2001)
Protein Eng. Des. Sel. 14, 939-942
   Abstract »    Full Text »    PDF »
Are the parameters of various stabilization factors estimated from mutant human lysozymes compatible with other proteins?.
J. Funahashi, K. Takano, and K. Yutani (2001)
Protein Eng. Des. Sel. 14, 127-134
   Abstract »    Full Text »    PDF »
Use of a non-rigid region in T4 lysozyme to design an adaptable metal-binding site.
J. W. Wray, W. A. Baase, G. J. Ostheimer, X.-j. Zhang, and B. W. Matthews (2000)
Protein Eng. Des. Sel. 13, 313-321
   Abstract »    Full Text »    PDF »
Structural characterization of an engineered tandem repeat contrasts the importance of context and sequence in protein folding.
M. Sagermann, W. A. Baase, and B. W. Matthews (1999)
PNAS 96, 6078-6083
   Abstract »    Full Text »    PDF »
Disulfide Bond Engineering to Monitor Conformational Opening of Apolipophorin III During Lipid Binding.
V. Narayanaswami, J. Wang, C. M. Kay, D. G. Scraba, and R. O. Ryan (1996)
J. Biol. Chem. 271, 26855-26862
   Abstract »    Full Text »    PDF »
GroEL Binds to and Unfolds Rhodanese Posttranslationally.
B. G. Reid and G. C. Flynn (1996)
J. Biol. Chem. 271, 7212-7217
   Abstract »    Full Text »    PDF »
The role of backbone flexibility in the accommodation of variants that repack the core of T4 lysozyme.
E. Baldwin, O Hajiseyedjavadi, W. Baase, and B. Matthews (1993)
Science 262, 1715-1718
   Abstract »    PDF »
ClpA mediates directional translocation of substrate proteins into the ClpP protease.
B. G. Reid, W. A. Fenton, A. L. Horwich, and E. U. Weber-Ban (2001)
PNAS 98, 3768-3772
   Abstract »    Full Text »    PDF »


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