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Shohdy <jats:italic>et al<\/jats:italic>. identified the <jats:italic>L. pneumophila<\/jats:italic> vacuole sorting inhibitory protein VipF as a putative <jats:italic>N<\/jats:italic>\u2010acetyltransferase based on sequence homology. We have characterized the basic structural and functional properties of VipF to confirm this original functional assignment. Sequence conservation analysis indicates two putative CoA\u2010binding regions within VipF. Homology modeling and small angle X\u2010ray scattering suggest a monomeric, dual\u2010domain structure joined by a flexible linker. Each domain contains the characteristic beta\u2010splay motif found in many acetyltransferases, suggesting that VipF may contain two active sites. Docking experiments suggest reasonable acetyl\u2010CoA binding locations within each beta\u2010splay motif. Broad substrate screening indicated that VipF is capable of acetylating chloramphenicol and both domains are catalytically active. Given that chloramphenicol is not known to be <jats:italic>N<\/jats:italic>\u2010acetylated, this is a surprising finding suggesting that VipF is capable of <jats:italic>O<\/jats:italic>\u2010acetyltransferase activity. Proteins 2016; 84:1422\u20131430. \u00a9 2016 Wiley Periodicals, Inc.<\/jats:p>","DOI":"10.1002\/prot.25087","type":"journal-article","created":{"date-parts":[[2016,6,17]],"date-time":"2016-06-17T13:21:05Z","timestamp":1466169665000},"page":"1422-1430","source":"Crossref","is-referenced-by-count":6,"title":["Characterization of the structure and catalytic activity of <i>Legionella pneumophila<\/i> VipF"],"prefix":"10.1002","volume":"84","author":[{"given":"Byron H.","family":"Young","sequence":"first","affiliation":[{"name":"Department of Chemistry and Biochemistry James Madison University Harrisonburg Virginia 22807"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tracy A.","family":"Caldwell","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry James Madison University Harrisonburg Virginia 22807"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aidan M.","family":"McKenzie","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry James Madison University Harrisonburg Virginia 22807"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Oleksandr","family":"Kokhan","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry James Madison University Harrisonburg Virginia 22807"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4285-4341","authenticated-orcid":false,"given":"Christopher E.","family":"Berndsen","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Biochemistry James Madison University Harrisonburg Virginia 22807"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2016,7,5]]},"reference":[{"key":"e_1_2_6_2_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0501315102"},{"key":"e_1_2_6_3_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.95.4.1669"},{"key":"e_1_2_6_4_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM197712012972201"},{"key":"e_1_2_6_5_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM197712012972202"},{"key":"e_1_2_6_6_1","doi-asserted-by":"publisher","DOI":"10.1078\/1438-4221-00154"},{"key":"e_1_2_6_7_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.158.4.1319"},{"key":"e_1_2_6_8_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1749-6632.2000.tb06177.x"},{"key":"e_1_2_6_9_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.129.2.321"},{"key":"e_1_2_6_10_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.ppat.1002546"},{"key":"e_1_2_6_11_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1316376111"},{"key":"e_1_2_6_12_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2958.1991.tb00770.x"},{"key":"e_1_2_6_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.abb.2004.09.003"},{"key":"e_1_2_6_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.sbi.2008.11.004"},{"key":"e_1_2_6_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mad.2010.05.001"},{"key":"e_1_2_6_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/jp8007514"},{"key":"e_1_2_6_17_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-890-0:571"},{"key":"e_1_2_6_18_1","doi-asserted-by":"publisher","DOI":"10.1110\/ps.051917406"},{"key":"e_1_2_6_19_1","doi-asserted-by":"publisher","DOI":"10.1021\/ed200060t"},{"key":"e_1_2_6_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.22570"},{"key":"e_1_2_6_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/pro.2199"},{"key":"e_1_2_6_22_1","doi-asserted-by":"crossref","unstructured":"KonarevPV VolkovVV SokolovaAV KochMHJ SvergunDI KochHJ.PRIMUS: a Windows PC\u2010based system for small\u2010angle scattering data analysis PRIMUS: a Windows PC\u2010based system for small\u2010 angle scattering data analysis.2003;36:1277\u20131282.","DOI":"10.1107\/S0021889803012779"},{"key":"e_1_2_6_23_1","doi-asserted-by":"publisher","DOI":"10.1107\/S002188989100081X"},{"key":"e_1_2_6_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0006-3495(99)77443-6"},{"key":"e_1_2_6_25_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889812015786"},{"key":"e_1_2_6_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.10349"},{"key":"e_1_2_6_27_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.20854"},{"key":"e_1_2_6_28_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.23062"},{"key":"e_1_2_6_29_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep08673"},{"key":"e_1_2_6_30_1","doi-asserted-by":"publisher","DOI":"10.1110\/ps.03153703"},{"key":"e_1_2_6_31_1","doi-asserted-by":"publisher","DOI":"10.1002\/prot.20967"},{"key":"e_1_2_6_32_1","doi-asserted-by":"crossref","unstructured":"FedorovAA. 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