Publication Details
HotSpot Wizard 2.0: Web Server for Automated Design of Mutations and Smart Libraries
Štourač Jan (LL)
Šebestová Eva, Mgr., Ph.D. (LL)
Vávra Ondřej, Bc. (LL)
Musil Miloš, Ing., Ph.D. (DIFS FIT BUT)
Brezovský Jan, Mgr., Ph.D. (LL)
Damborský Jiří, prof. Mgr., Dr. (LL)
protein engineering, smart library design, computational mutagenesis, hotspot prediction, semi-rational enzyme design
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.
@ARTICLE{FITPUB11083, author = "Jaroslav Bendl and Jan \v{S}toura\v{c} and Eva \v{S}ebestov\'{a} and Ond\v{r}ej V\'{a}vra and Milo\v{s} Musil and Jan Brezovsk\'{y} and Ji\v{r}\'{i} Damborsk\'{y}", title = "HotSpot Wizard 2.0: Web Server for Automated Design of Mutations and Smart Libraries", pages = "479--487", journal = "Nucleic Acids Research", volume = 44, number = 1, year = 2016, ISSN = "1362-4962", doi = "10.1093/nar/gkp410", language = "english", url = "https://www.fit.vut.cz/research/publication/11083" }