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@article{1322952,
author = {Bednář, David and Beerens, Koen and Šebestová, Eva and Bendl, Jaroslav and Khare, S. and Chaloupková, Radka and Prokop, Zbyněk and Brezovský, Jan and Baker, D. and Damborský, Jiří},
article_number = {11},
doi = {http://dx.doi.org/10.1371/journal.pcbi.1004556},
keywords = {FireProt;haloalkane dehalogenase},
language = {eng},
issn = {1553-734X},
journal = {PLOS COMPUTATIONAL BIOLOGY},
title = {FireProt: Energy- and Evolution-Based Computational Design of Thermostable Multiple-Point Mutants},
url = {http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/11/Bednar_2015ploscb1.pdf},
volume = {11},
year = {2015}
}
TY - JOUR
ID - 1322952
AU - Bednář, David - Beerens, Koen - Šebestová, Eva - Bendl, Jaroslav - Khare, S. - Chaloupková, Radka - Prokop, Zbyněk - Brezovský, Jan - Baker, D. - Damborský, Jiří
PY - 2015
TI - FireProt: Energy- and Evolution-Based Computational Design of Thermostable Multiple-Point Mutants
JF - PLOS COMPUTATIONAL BIOLOGY
VL - 11
IS - 11
SP - "nestrankovano"
EP - "nestrankovano"
SN - 1553734X
KW - FireProt;haloalkane dehalogenase
UR - http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/11/Bednar_2015ploscb1.pdf
L2 - http://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2015/11/Bednar_2015ploscb1.pdf
N2 - There is great interest in increasing proteins’ stability to enhance their utility as biocatalysts, therapeutics, diagnostics and nanomaterials. 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 and then recombined in multiple-point mutants. Thus, substantial screening is still required. Here we present FireProt, a robust computational strategy for predicting highly stable multiple-point mutants that combines energy- and evolution-based approaches with smart filtering to identify additive stabilizing mutations. FireProt’s reliability and applicability was demonstrated by validating its predictions against 656 mutations from the ProTherm database. We demonstrate that thermostability of the model enzymes haloalkane dehalogenase DhaA and gama-hexachlorocyclohexane dehydrochlorinase LinA can be substantially increased (Tm = 24°C and 21°C) by constructing and characterizing only a handful of multiple-point mutants. FireProt can be applied to any protein for which a tertiary structure and homologous sequences are available, and will facilitate the rapid development of robust proteins for biomedical and biotechnological applications.
ER -
BEDNÁŘ, David, Koen BEERENS, Eva ŠEBESTOVÁ, Jaroslav BENDL, S. KHARE, Radka CHALOUPKOVÁ, Zbyněk PROKOP, Jan BREZOVSKÝ, D. BAKER and Jiří DAMBORSKÝ. FireProt: Energy- and Evolution-Based Computational Design of Thermostable Multiple-Point Mutants. \textit{PLOS COMPUTATIONAL BIOLOGY}. 2015, vol.~11, No~11, p.~''nestrankovano'', 20 pp. ISSN~1553-734X. Available from: https://dx.doi.org/10.1371/journal.pcbi.1004556.
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