Publication details

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

Authors

BENDL J. ŠTOURAČ Jan ŠEBESTOVÁ Eva VÁVRA Ondřej MUSIL Miloš BREZOVSKÝ Jan DAMBORSKÝ Jiří

Year of publication 2016
Type Article in Periodical
Magazine / Source Nucleic Acids Research
MU Faculty or unit

Faculty of Science

Citation
Web https://loschmidt.chemi.muni.cz/peg/wp-content/uploads/2016/05/Bendl_2016NAR.pdf
Doi http://dx.doi.org/10.1093/nar/gkw416
Field Biochemistry
Keywords MULTIPLE SEQUENCE ALIGNMENTS; FUNCTIONALLY IMPORTANT RESIDUES; FOCUSED DIRECTED EVOLUTION; SATURATION MUTAGENESIS; CORRELATED MUTATIONS; ENZYME PROPERTIES; THERMOSTABILITY; IDENTIFICATION; CONSERVATION; BINDING
Description HotSpot Wizard 2.0 is a web server for automated identification of hot spots and design of smart libraries for engineering proteins’ stability, catalytic activity, substrate specificity and enantioselectivity. The server integrates sequence, structural and evolutionary information obtained from 3 databases and 20 computational tools. Users are guided through the processes of selecting hot spots using four different protein engineering strategies and optimizing the resulting library’s size by narrowing down a set of substitutions at individual randomized positions. The only required input is a query protein structure. The results of the calculations are mapped onto the protein’s structure and visualized with a JSmol applet. HotSpot Wizard lists annotated residues suitable for mutagenesis and can automatically design appropriate codons for each implemented strategy. Overall, HotSpot Wizard provides comprehensive annotations of protein structures and assists protein engineers with the rational design of site-specific mutations and focused libraries. It is freely available at http://loschmidt.chemi.muni.cz/hotspotwizard.
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