{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T17:34:10Z","timestamp":1782408850825,"version":"3.54.5"},"reference-count":50,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2014,4,9]],"date-time":"2014-04-09T00:00:00Z","timestamp":1397001600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"ANCyPT"},{"DOI":"10.13039\/501100002923","name":"CONICET","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100002923","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2014,5]]},"abstract":"<jats:sec><jats:label\/><jats:p>Cerulenin is a fungal toxin that inhibits both eukaryotic and prokaryotic ketoacyl\u2010acyl carrier protein synthases or condensing enzymes. It has been used experimentally to treat cancer and obesity, and is a potent inhibitor of bacterial growth. Understanding the molecular mechanisms of resistance to cerulenin and similar compounds is thus highly relevant for human health. We have previously described a <jats:italic><jats:styled-content style=\"fixed-case\">B<\/jats:styled-content>acillus\u00a0subtilis<\/jats:italic> cerulenin\u2010resistant strain, expressing a point\u2010mutated condensing enzyme FabF (FabF[I108F]) (i.e. FabF with isoleucine 108 substituted by phenylalanine). We now report the crystal structures of wild\u2010type FabF from <jats:italic>B.\u00a0subtilis<\/jats:italic>, both alone and in complex with cerulenin, as well as of the FabF[I108F] mutant protein. The three\u2010dimensional structure of FabF[I108F] constitutes the first atomic model of a condensing enzyme that remains active in the presence of the inhibitor. Soaking the mycotoxin into preformed wild\u2010type FabF crystals allowed for noncovalent binding into its specific pocket within the FabF core. Interestingly, only co\u2010crystallization experiments allowed us to trap the covalent complex. Our structure shows that the covalent bond between Cys163 and cerulenin, in contrast to that previously proposed, implicates carbon C3 of the inhibitor. The similarities between <jats:italic><jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>scherichia\u00a0coli<\/jats:italic> and <jats:italic><jats:styled-content style=\"fixed-case\">B<\/jats:styled-content>.\u00a0subtilis<\/jats:italic> FabF structures did not explain the reported inability of ecFabF[I108F] (i.e. FabF from <jats:italic><jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>scherichia\u00a0coli<\/jats:italic> with isoleucine 108 substituted by phenylalanine) to elongate medium and long\u2010chain acyl\u2010<jats:styled-content style=\"fixed-case\">ACP<\/jats:styled-content>s. We now demonstrate that the <jats:italic><jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>.\u00a0coli<\/jats:italic> modified enzyme efficiently catalyzes the synthesis of medium and long\u2010chain ketoacyl\u2010<jats:styled-content style=\"fixed-case\">ACP<\/jats:styled-content>s. We also characterized another cerulenin\u2010insensitive form of FabF, conferring a different phenotype in <jats:italic><jats:styled-content style=\"fixed-case\">B<\/jats:styled-content>.\u00a0subtilis<\/jats:italic>. The structural, biochemical and physiological data presented, shed light on the mechanisms of FabF catalysis and resistance to cerulenin.<\/jats:p><\/jats:sec><jats:sec><jats:title>Database<\/jats:title><jats:p>Crystallographic data (including atomic coordinates and structure factors) have been deposited in the Protein Data Bank under accession codes <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=4LS5\">4LS5<\/jats:ext-link>, <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=4LS6\">4LS6<\/jats:ext-link>, <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=4LS7\">4LS7<\/jats:ext-link> and <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.rcsb.org\/pdb\/search\/structidSearch.do?structureId=4LS8\">4LS8<\/jats:ext-link>.<\/jats:p><\/jats:sec>","DOI":"10.1111\/febs.12785","type":"journal-article","created":{"date-parts":[[2014,3,19]],"date-time":"2014-03-19T05:09:01Z","timestamp":1395205741000},"page":"2324-2338","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":44,"title":["Structural insights into bacterial resistance to cerulenin"],"prefix":"10.1111","volume":"281","author":[{"given":"Felipe","family":"Trajtenberg","sequence":"first","affiliation":[{"name":"Institut Pasteur de Montevideo Unit of Protein Crystallography Montevideo Uruguay"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Silvia","family":"Altabe","sequence":"additional","affiliation":[{"name":"Instituto de Biolog\u00eda Molecular y Celular de Rosario (IBR) \u2013 CONICET Facultad de Cs Bioqu\u00edmicas y Farmac\u00e9uticas Universidad Nacional de Rosario  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