{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,17]],"date-time":"2026-03-17T07:06:03Z","timestamp":1773731163024,"version":"3.50.1"},"reference-count":16,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2012,4,24]],"date-time":"2012-04-24T00:00:00Z","timestamp":1335225600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2012,6]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:sc>D<\/jats:sc>\u2010Tetronitrose is a nitro\u2010containing tetradeoxysugar found attached to the antitumor and antibacterial agent tetrocarcin A. The biosynthesis of this highly unusual sugar in <jats:italic>Micromonospora chalcea<\/jats:italic> requires 10 enzymes. The fifth step in the pathway involves the transfer of a methyl group from <jats:italic>S<\/jats:italic>\u2010adenosyl\u2010<jats:sc>L<\/jats:sc>\u2010methionine (SAM) to the C\u20103\u2032 carbon of dTDP\u20103\u2010amino\u20102,3,6\u2010trideoxy\u20104\u2010keto\u2010<jats:sc>D<\/jats:sc>\u2010glucose. The enzyme responsible for this transformation is referred to as TcaB9. It is a monomeric enzyme with a molecular architecture based around three domains. The N\u2010terminal motif contains a binding site for a structural zinc ion. The middle\u2010 and C\u2010terminal domains serve to anchor the SAM and dTDP\u2013sugar ligands, respectively, to the protein, and the active site of TcaB9 is wedged between these two regions. For this investigation, the roles of Tyr 76, His 181, Tyr 222, Glu 224, and His 225, which form the active site of TcaB9, were probed by site\u2010directed mutagenesis, kinetic analyses, and X\u2010ray structural studies. In addition, two ternary complexes of the enzyme with bound <jats:italic>S<\/jats:italic>\u2010adenosyl\u2010<jats:sc>L<\/jats:sc>\u2010homocysteine and either dTDP\u20103\u2010amino\u20102,3,6\u2010trideoxy\u20104\u2010keto\u2010<jats:sc>D<\/jats:sc>\u2010glucose or dTDP\u20103\u2010amino\u20102,3,6\u2010trideoxy\u2010<jats:sc>D<\/jats:sc>\u2010galactose were determined to 1.5 or 1.6 \u00c5 resolution, respectively. Taken together, these investigations highlight the important role of His 225 in methyl transfer. In addition, the structural data suggest that the methylation reaction occurs via retention of configuration about the C\u20103\u2032 carbon of the sugar.<\/jats:p>","DOI":"10.1002\/pro.2074","type":"journal-article","created":{"date-parts":[[2012,4,12]],"date-time":"2012-04-12T01:15:05Z","timestamp":1334193305000},"page":"876-886","source":"Crossref","is-referenced-by-count":6,"title":["Probing the catalytic mechanism of a <i>C<\/i>\u20103\u2032\u2010methyltransferase involved in the biosynthesis of <scp>D<\/scp>\u2010tetronitrose"],"prefix":"10.1002","volume":"21","author":[{"given":"Nathan A.","family":"Bruender","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hazel M.","family":"Holden","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2012,4,24]]},"reference":[{"key":"e_1_2_5_2_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.33.668"},{"key":"e_1_2_5_3_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.35.1033"},{"key":"e_1_2_5_4_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.36.595"},{"key":"e_1_2_5_5_2","doi-asserted-by":"publisher","DOI":"10.1128\/JB.00533-08"},{"key":"e_1_2_5_6_2","doi-asserted-by":"publisher","DOI":"10.7164\/antibiotics.35.979"},{"key":"e_1_2_5_7_2","doi-asserted-by":"publisher","DOI":"10.1021\/ja0744854"},{"key":"e_1_2_5_8_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi100782b"},{"key":"e_1_2_5_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/1521-3773(20010202)40:3<607::AID-ANIE607>3.0.CO;2-8"},{"key":"e_1_2_5_10_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.210395097"},{"key":"e_1_2_5_11_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi100318v"},{"key":"e_1_2_5_12_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889807021206"},{"key":"e_1_2_5_13_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"e_1_2_5_14_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444996012255"},{"key":"e_1_2_5_15_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0021889892009944"},{"key":"e_1_2_5_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0076-6879(79)63008-2"},{"key":"e_1_2_5_17_2","unstructured":"DeLanoWL(2002)The PyMOL Molecular Graphics System. 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DeLano Scientific San Carlos CA USA."}],"container-title":["Protein Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fpro.2074","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pro.2074","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,10]],"date-time":"2023-10-10T00:29:52Z","timestamp":1696897792000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/pro.2074"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,4,24]]},"references-count":16,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2012,6]]}},"alternative-id":["10.1002\/pro.2074"],"URL":"https:\/\/doi.org\/10.1002\/pro.2074","archive":["Portico"],"relation":{},"ISSN":["0961-8368","1469-896X"],"issn-type":[{"value":"0961-8368","type":"print"},{"value":"1469-896X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,4,24]]}}}