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Research efforts to identify antigenic components for a melioidosis vaccine have led to the identification of several proteins, including subunits forming the flagella that mediate bacterial motility, host colonization, and virulence. This study focuses on the <jats:italic>B.\u00a0pseudomallei<\/jats:italic> flagellar hook\u2010associated protein (Flg<jats:styled-content style=\"fixed-case\">K<jats:sub>B<\/jats:sub><\/jats:styled-content><jats:sub>p<\/jats:sub>), and provides the first insights into the 3D structure of FlgK proteins as targets for structure\u2010based antigen engineering. The Flg<jats:styled-content style=\"fixed-case\">K<jats:sub>B<\/jats:sub><\/jats:styled-content><jats:sub>p<\/jats:sub> crystal structure (presented here at 1.8\u2010\u00c5 resolution) reveals a multidomain fold, comprising two small \u03b2\u2010domains protruding from a large elongated \u03b1\u2010helical bundle core. The evident structural similarity to flagellin, the flagellar filament subunit protein, suggests that, depending on the bacterial species, flagellar hook\u2010associated proteins are likely to show a conserved, elongated \u03b1\u2010helical bundle scaffold coupled to a variable number of smaller domains. Furthermore, we present immune serum recognition data confirming, in agreement with previous findings, that recovered melioidosis patients produce elevated levels of antibodies against Flg<jats:styled-content style=\"fixed-case\">K<jats:sub>B<\/jats:sub><\/jats:styled-content><jats:sub>p<\/jats:sub>, in comparison with seronegative and seropositive healthy subjects. Moreover, we show that Flg<jats:styled-content style=\"fixed-case\">K<jats:sub>B<\/jats:sub><\/jats:styled-content><jats:sub>p<\/jats:sub> has cytotoxic effects on cultured murine macrophages, suggesting an important role in bacterial pathogenesis. Finally, computational epitope prediction methods applied to the Flg<jats:styled-content style=\"fixed-case\">K<jats:sub>B<\/jats:sub><\/jats:styled-content><jats:sub>p<\/jats:sub> crystal structure, coupled with <jats:italic>in\u00a0vitro<\/jats:italic> mapping, allowed us to predict three antigenic regions that locate to discrete protein domains. Taken together, our results point to Flg<jats:styled-content style=\"fixed-case\">K<jats:sub>B<\/jats:sub><\/jats:styled-content><jats:sub>p<\/jats:sub> as a candidate for the design and production of epitope\u2010containing subunits\/domains as potential vaccine components.<\/jats:p>","DOI":"10.1111\/febs.13223","type":"journal-article","created":{"date-parts":[[2015,2,3]],"date-time":"2015-02-03T06:00:17Z","timestamp":1422943217000},"page":"1319-1333","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":40,"title":["From crystal structure to <i>in\u00a0silico<\/i> epitope discovery in the <i>Burkholderia\u00a0pseudomallei<\/i> flagellar hook\u2010associated protein FlgK"],"prefix":"10.1111","volume":"282","author":[{"given":"Louise J.","family":"Gourlay","sequence":"first","affiliation":[{"name":"Department of Biosciences University of Milan Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachael J.","family":"Thomas","sequence":"additional","affiliation":[{"name":"College of Life and Environmental Sciences University of Exeter UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Claudio","family":"Peri","sequence":"additional","affiliation":[{"name":"College of Life and Environmental Sciences University of Exeter UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oscar","family":"Conchillo\u2010Sol\u00e9","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology and Biomedicine Universitat Aut\u00f2noma de Barcelona Bellaterra Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mario","family":"Ferrer\u2010Navarro","sequence":"additional","affiliation":[{"name":"IsGlobal Barcelona Ctr. 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