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Nucleic Acids Research Advance Access originally published online on May 21, 2009
Nucleic Acids Research 2009 37(Web Server issue):W376-W383; doi:10.1093/nar/gkp410
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Nucleic Acids Research, 2009, Vol. 37, No. suppl_2 W376-W383
© 2009 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Articles

HotSpot Wizard: a web server for identification of hot spots in protein engineering

Antonin Pavelka, Eva Chovancova and Jiri Damborsky*

Loschmidt Laboratories, Institute of Experimental Biology and National Centre for Biomolecular Research, Faculty of Science, Masaryk University, Kamenice 5/A4, 625 00 Brno, Czech Republic

*To whom correspondence should be addressed. Tel: +420 5 49493467; Fax: +420 5 49496302; Email: jiri{at}chemi.muni.cz

Received February 18, 2009. Revised April 18, 2009. Accepted May 2, 2009.

HotSpot Wizard is a web server for automatic identification of ‘hot spots’ for engineering of substrate specificity, activity or enantioselectivity of enzymes and for annotation of protein structures. The web server implements the protein engineering protocol, which targets evolutionarily variable amino acid positions located in the active site or lining the access tunnels. The ‘hot spots’ for mutagenesis are selected through the integration of structural, functional and evolutionary information obtained from: (i) the databases RCSB PDB, UniProt, PDBSWS, Catalytic Site Atlas and nr NCBI and (ii) the tools CASTp, CAVER, BLAST, CD-HIT, MUSCLE and Rate4Site. The protein structure and e-mail address are the only obligatory inputs for the calculation. In the output, HotSpot Wizard lists annotated residues ordered by estimated mutability. The results of the analysis are mapped on the enzyme structure and visualized in the web browser using Jmol. The HotSpot Wizard server should be useful for protein engineers interested in exploring the structure of their favourite protein and for the design of mutations in site-directed mutagenesis and focused directed evolution experiments. HotSpot Wizard is available at http://loschmidt.chemi.muni.cz/hotspotwizard/.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors.


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