Publication Details

FireProt:Energy- and evolution-based computational design of thermostable multiple-point mutants

BEDNÁŘ David, BEERENS Koen, ŠEBESTOVÁ Eva, BENDL Jaroslav, KHARE Sagar, CHALOUPKOVÁ Radka, PROKOP Zbyněk, BREZOVSKÝ Jan, BAKER David and DAMBORSKÝ Jiří. FireProt:Energy- and evolution-based computational design of thermostable multiple-point mutants. PLoS Computational Biology, vol. 11, no. 11, 2015, pp. 1-20. ISSN 1553-7358. Available from: http://www.ploscompbiol.org/article/fetchObject.action?uri=info:doi/10.1371/journal.pcbi.1004556&representation=PDF
Czech title
FireProt: Výpočetní design více-bodového termostabilního proteinu
Type
journal article
Language
english
Authors
Bednář David, Mgr. (LL)
Beerens Koen, Ph.D. (LL)
Šebestová Eva, Mgr., Ph.D. (LL)
Bendl Jaroslav, Ing. (DIFS FIT BUT)
Khare Sagar, Ph.D. (RUTGERS)
Chaloupková Radka, Mgr., Ph.D. (LL)
Prokop Zbyněk, doc. RnDr., Ph.D. (LL)
Brezovský Jan, Mgr., Ph.D. (LL)
Baker David, prof., Ph.D. (UWASH)
Damborský Jiří, prof. Mgr., Dr. (LL)
URL
Keywords

protein stability
protein thermostability
improvement of enzymatic properties
multi-point mutants

Abstract

There is great interest in increasing proteins' stability to enhance their utility as biocatalysts, therapeutics, diagnostics and nano-materials. Directed evolution is a powerful, but experimentally strenuous approach. Computational methods offer attractive alternatives. However, due to the limited reliability of predictions and potentially antagonistic effects of substitutions, only single-point mutations are usually predicted in silico, experimentally verified, then recombined in multiple-point mutants. Thus, substantial screening is still required. Here we present a robust computational strategy for predicting highly stable multiple-point mutants, called FireProt, combining energy- and evolution-based approaches with smart filtering to identify additive stabilizing mutations. We show that thermostability of the model enzyme haloalkane dehalogenase DhaA can be substantially increased (dTm , 24 o C) by constructing and characterizing as few as six multiple-point mutants. The method is generally applicable to all proteins with known tertiary structure and homologous sequences, and should facilitate rapid development of robust proteins for biomedical and biotechnological applications.

Published
2015
Pages
1-20
Journal
PLoS Computational Biology, vol. 11, no. 11, ISSN 1553-7358
Publisher
Public Library of Science
DOI
10.1371/journal.pcbi.1004556
UT WoS
000365801600026
EID Scopus
2-s2.0-84949257169
BibTeX 
@ARTICLE{FITPUB10784,
   author = "David Bedn\'{a}\v{r} and Koen Beerens and Eva \v{S}ebestov\'{a} and Jaroslav Bendl and Sagar Khare and Radka Chaloupkov\'{a} and Zbyn\v{e}k Prokop and Jan Brezovsk\'{y} and David Baker and Ji\v{r}\'{i} Damborsk\'{y}",
   title = "FireProt:Energy- and evolution-based computational design of thermostable multiple-point mutants",
   pages = "1--20",
   journal = "PLoS Computational Biology",
   volume = 11,
   number = 11,
   year = 2015,
   ISSN = "1553-7358",
   doi = "10.1371/journal.pcbi.1004556",
   language = "english",
   url = "https://www.fit.vut.cz/research/publication/10784"
}
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