Related Content
Search Google Scholar for:
|
|
Science 6 May 2005:
Vol. 308. no. 5723, pp. 857 - 860
DOI: 10.1126/science.1107387
|
|
Reports
Computational Thermostabilization of an Enzyme
Aaron Korkegian,1,2
Margaret E. Black,4
David Baker,3
Barry L. Stoddard1*
Thermostabilizing an enzyme while maintaining its activity for industrial or biomedical applications can be difficult with traditional selection methods. We describe a rapid computational approach that identified three mutations within a model enzyme that produced a 10°C increase in apparent melting temperature Tm and a 30-fold increase in half-life at 50°C, with no reduction in catalytic efficiency. The effects of the mutations were synergistic, giving an increase in excess of the sum of their individual effects. The redesigned enzyme induced an increased, temperature-dependent bacterial growth rate under conditions that required its activity, thereby coupling molecular and metabolic engineering.
1 Division of Basic Sciences, Fred Hutchinson Cancer Research Center (FHCRC), 1100 Fairview Avenue North, Seattle, WA 98109, USA.
2 Graduate Program in Molecular and Cellular Biology, University of Washington, Seattle, WA 98195, USA.
3 Department of Biochemistry and Howard Hughes Medical Institute, University of Washington, Seattle, WA 98195, USA.
4 Department of Pharmaceutical Sciences, College of Pharmacy, Washington State University, Pullman, WA 99164–6534, USA.
* To whom correspondence should be addressed. E-mail: bstoddar{at}fhcrc.org
Read the Full Text
THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
- Optimal design of thermally stable proteins.
- R. M. Bannen, V. Suresh, G. N. Phillips Jr, S. J. Wright, and J. C. Mitchell (2008)
Bioinformatics
24, 2339-2343
| Abstract »
| Full Text »
| PDF »
- Thermostable variants constructed via the structure-guided consensus method also show increased stability in salts solutions and homogeneous aqueous-organic media.
- E. Vazquez-Figueroa, V. Yeh, J.M. Broering, J.F. Chaparro-Riggers, and A.S. Bommarius (2008)
Protein Eng. Des. Sel.
| Abstract »
| Full Text »
| PDF »
- Lysine acetylation can generate highly charged enzymes with increased resistance toward irreversible inactivation.
- B. F. Shaw, G. F. Schneider, B. Bilgicer, G. K. Kaufman, J. M. Neveu, W. S. Lane, J. P. Whitelegge, and G. M. Whitesides (2008)
Protein Sci.
17, 1446-1455
| Abstract »
| Full Text »
| PDF »
- Bioinformatic method for protein thermal stabilization by structural entropy optimization.
- E. Bae, R. M. Bannen, and G. N. Phillips Jr. (2008)
PNAS
105, 9594-9597
| Abstract »
| Full Text »
| PDF »
- Algorithm for backrub motions in protein design.
- I. Georgiev, D. Keedy, J. S. Richardson, D. C. Richardson, and B. R. Donald (2008)
Bioinformatics
24, i196-i204
| Abstract »
| Full Text »
| PDF »
- Generation and Analysis of Mesophilic Variants of the Thermostable Archaeal I-DmoI Homing Endonuclease.
- J. Prieto, J.-C. Epinat, P. Redondo, E. Ramos, D. Padro, F. Cedrone, G. Montoya, F. Paques, and F. J. Blanco (2008)
J. Biol. Chem.
283, 4364-4374
| Abstract »
| Full Text »
| PDF »
- Computational design of the Fyn SH3 domain with increased stability through optimization of surface charge charge interactions.
- K. L. Schweiker, A. Zarrine-Afsar, A. R. Davidson, and G. I. Makhatadze (2007)
Protein Sci.
16, 2694-2702
| Abstract »
| Full Text »
| PDF »
- Four-Body Scoring Function for Mutagenesis.
- C. Deutsch and B. Krishnamoorthy (2007)
Bioinformatics
23, 3009-3015
| Abstract »
| Full Text »
| PDF »
- Optimal protein library design using recombination or point mutations based on sequence-based scoring functions.
- R. J. Pantazes, M. C. Saraf, and C. D. Maranas (2007)
Protein Eng. Des. Sel.
| Abstract »
| Full Text »
| PDF »
- Stabilization of Hyperactive Dihydrofolate Reductase by Cyanocysteine-mediated Backbone Cyclization.
- H. Takahashi, M. Arai, T. Takenawa, H. Sota, Q. H. Xie, and M. Iwakura (2007)
J. Biol. Chem.
282, 9420-9429
| Abstract »
| Full Text »
| PDF »
- Extending Iterative Protein Redesign and Optimization (IPRO) in Protein Library Design for Ligand Specificity.
- H. Fazelinia, P. C. Cirino, and C. D. Maranas (2007)
Biophys. J.
92, 2120-2130
| Abstract »
| Full Text »
| PDF »
- Rapid detection of similarity in protein structure and function through contact metric distances.
- A. M. Lisewski and O. Lichtarge (2006)
Nucleic Acids Res.
34, e152
| Abstract »
| Full Text »
| PDF »
- Bioinformatics-driven, rational engineering of protein thermostability.
- M. K. DiTursi, S.-J. Kwon, P. J. Reeder, and J. S. Dordick (2006)
Protein Eng. Des. Sel.
19, 517-524
| Abstract »
| Full Text »
| PDF »
- Lessons in stability from thermophilic proteins..
- A. Razvi and J. M. Scholtz (2006)
Protein Sci.
15, 1569-1578
| Abstract »
| Full Text »
| PDF »
- RosettaDesign server for protein design..
- Y. Liu and B. Kuhlman (2006)
Nucleic Acids Res.
34, W235-W238
| Abstract »
| Full Text »
| PDF »
- IPRO: An Iterative Computational Protein Library Redesign and Optimization Procedure.
- M. C. Saraf, G. L. Moore, N. M. Goodey, V. Y. Cao, S. J. Benkovic, and C. D. Maranas (2006)
Biophys. J.
90, 4167-4180
| Abstract »
| Full Text »
| PDF »
- Comparison of crystal structure interactions and thermodynamics for stabilizing mutations in the Tetrahymena ribozyme.
- F. GUO, A. R. GOODING, and T. R. CECH (2006)
RNA
12, 387-395
| Abstract »
| Full Text »
| PDF »
- A Stability Pattern of Protein Hydrophobic Mutations that Reflects Evolutionary Structural Optimization.
- R. Godoy-Ruiz, R. Perez-Jimenez, B. Ibarra-Molero, and J. M. Sanchez-Ruiz (2005)
Biophys. J.
89, 3320-3331
| Abstract »
| Full Text »
| PDF »
|
|
