Publication detail

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.

Original Title

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

Type

journal article in Web of Science

Language

English

Original 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.

Keywords

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

Authors

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

RIV year

2015

Released

3. 11. 2015

ISBN

1553-7358

Periodical

PLoS Computational Biology

Year of study

11

Number

11

State

United States of America

Pages from

1

Pages to

20

Pages count

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"
}