Publication detail

HotSpot Wizard 2.0: Automated Design of Site-specific Mutations and Smart Libraries in Protein Engineering

BENDL, J. ŠTOURAČ, J. ŠEBESTOVÁ, E. VÁVRA, O. MUSIL, M. BREZOVSKÝ, J. DAMBORSKÝ, J.

Original Title

HotSpot Wizard 2.0: Automated Design of Site-specific Mutations and Smart Libraries in Protein Engineering

Type

journal article in Web of Science

Language

English

Original Abstract

HotSpot Wizard 2.0 is a web server for an automatic identification of hot spots and design of mutations for engineering of enzyme activity, substrate specificity, and enantioselectivity or protein stability. The server integrates sequence, structural, and evolutionary information obtained from three databases and nineteen computational tools to guide the users through the process of hot spot selection by four protein engineering strategies as well as the optimization of library size by narrowing down a set of substitutions at individual randomized positions. The protein structure represents the only obligatory input for the calculation. The results of calculations are mapped on the protein structure and visualized in JSMol applet. For each implemented strategy, HotSpot Wizard lists annotated residues suitable for mutagenesis and enables to design appropriate codons. Altogether, HotSpot Wizard provides a comprehensive annotation of protein structures and assists protein engineers with the design of mutations in site-directed mutagenesis and focused directed evolution experiments. HotSpot Wizard is freely available at http://loschmidt.chemi.muni.cz/hotspotwizard2.

Keywords

protein engineering, smart library design, computational mutagenesis, hotspot prediction, semi-rational enzyme design

Authors

BENDL, J.; ŠTOURAČ, J.; ŠEBESTOVÁ, E.; VÁVRA, O.; MUSIL, M.; BREZOVSKÝ, J.; DAMBORSKÝ, J.

Released

13. 5. 2016

ISBN

1362-4962

Periodical

Nucleic Acids Research

Year of study

44

Number

1

State

United Kingdom of Great Britain and Northern Ireland

Pages from

479

Pages to

487

Pages count

9

URL

http://nar.oxfordjournals.org/content/early/2016/05/12/nar.gkw416.full

BibTex

@article{BUT133492,
  author="Jaroslav {Bendl} and Jan {Štourač} and Eva {Chovancová} and Ondřej {Vávra} and Miloš {Musil} and Jan {Brezovský} and Jiří {Damborský}",
  title="HotSpot Wizard 2.0: Automated Design of Site-specific Mutations and Smart Libraries in Protein Engineering",
  journal="Nucleic Acids Research",
  year="2016",
  volume="44",
  number="1",
  pages="479--487",
  doi="10.1093/nar/gkp410",
  issn="1362-4962",
  url="http://nar.oxfordjournals.org/content/early/2016/05/12/nar.gkw416.full"
}