Detail publikace

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

BEDNÁŘ, D. BEERENS, K. ŠEBESTOVÁ, E. BENDL, J. KHARE, S. CHALOUPKOVÁ, R. PROKOP, Z. BREZOVSKÝ, J. BAKER, D. DAMBORSKÝ, J.

Originální název

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

Typ

článek v časopise ve Web of Science, Jimp

Jazyk

angličtina

Originální abstrakt

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.

Klíčová slova

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

Autoři

BEDNÁŘ, D.; BEERENS, K.; ŠEBESTOVÁ, E.; BENDL, J.; KHARE, S.; CHALOUPKOVÁ, R.; PROKOP, Z.; BREZOVSKÝ, J.; BAKER, D.; DAMBORSKÝ, J.

Rok RIV

2015

Vydáno

3. 11. 2015

ISSN

1553-7358

Periodikum

PLoS Computational Biology

Ročník

11

Číslo

11

Stát

Spojené státy americké

Strany od

1

Strany do

20

Strany počet

20

URL

http://www.ploscompbiol.org/article/fetchObject.action?uri=info:doi/10.1371/journal.pcbi.1004556&representation=PDF

BibTex

@article{BUT133486,
  author="David {Bednář} and Koen {Beerens} and Eva {Chovancová} and Jaroslav {Bendl} and Sagar {Khare} and Radka {Chaloupková} and Zbyněk {Prokop} and Jan {Brezovský} and David {Baker} and Jiří {Damborský}",
  title="FireProt:Energy- and evolution-based computational design of thermostable multiple-point mutants",
  journal="PLoS Computational Biology",
  year="2015",
  volume="11",
  number="11",
  pages="1--20",
  doi="10.1371/journal.pcbi.1004556",
  issn="1553-7358",
  url="http://www.ploscompbiol.org/article/fetchObject.action?uri=info:doi/10.1371/journal.pcbi.1004556&representation=PDF"
}