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The bioinformatic analysis is facilitated by Mustguseal\u2014a companion web-server to automatically collect and superimpose a large representative set of functionally diverse homologs with high structure similarity but low sequence identity to the selected query protein. The results are automatically prioritized and provided at four information levels to facilitate the knowledge-driven expert selection of the most promising positions on-line: as a sequence similarity network; interfaces to sequence-based and 3D-structure-based analysis of conservation and variability; and accompanied by the detailed annotation of proteins accumulated from the integrated databases with links to the external resources. The integration of Zebra2 and Mustguseal web-tools provides the first of its kind out-of-the-box open-access solution to conduct a systematic analysis of evolutionarily related proteins implementing different functions within a shared 3D-structure of the superfamily, determine common and specific patterns of function-associated local structural elements, assist to select hot-spots for rational design and to prepare focused libraries for directed evolution. The web-servers are free and open to all users at https:\/\/biokinet.belozersky.msu.ru\/zebra2, no login required.<\/jats:p>","DOI":"10.1093\/nar\/gkaa276","type":"journal-article","created":{"date-parts":[[2020,4,8]],"date-time":"2020-04-08T19:09:55Z","timestamp":1586372995000},"page":"W65-W71","source":"Crossref","is-referenced-by-count":20,"title":["Zebra2: advanced and easy-to-use web-server for bioinformatic analysis of subfamily-specific and conserved positions in diverse protein superfamilies"],"prefix":"10.1093","volume":"48","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7054-3487","authenticated-orcid":false,"given":"Dmitry","family":"Suplatov","sequence":"first","affiliation":[{"name":"Lomonosov Moscow State University, Belozersky Institute of Physicochemical Biology and Faculty of Bioengineering and Bioinformatics, Lenin Hills\u00a01-73, Moscow 119234, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yana","family":"Sharapova","sequence":"first","affiliation":[{"name":"Lomonosov Moscow State University, Belozersky Institute of Physicochemical Biology and Faculty of Bioengineering and Bioinformatics, Lenin Hills\u00a01-73, Moscow 119234, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Elizaveta","family":"Geraseva","sequence":"first","affiliation":[{"name":"Lomonosov Moscow State University, Belozersky Institute of Physicochemical Biology and Faculty of Bioengineering and Bioinformatics, Lenin Hills\u00a01-73, Moscow 119234, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vytas","family":"\u0160vedas","sequence":"first","affiliation":[{"name":"Lomonosov Moscow State University, Belozersky Institute of Physicochemical Biology and Faculty of Bioengineering and Bioinformatics, Lenin Hills\u00a01-73, Moscow 119234, Russia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2020,4,20]]},"reference":[{"key":"2020062614050878600_B1","doi-asserted-by":"crossref","first-page":"689","DOI":"10.1093\/protein\/gzs068","article-title":"Bioinformatic analysis of alpha\/beta-hydrolase fold enzymes reveals subfamily-specific positions responsible for discrimination of amidase and lipase activities","volume":"25","author":"Suplatov","year":"2012","journal-title":"Protein Eng. 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