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Although previously implicated to employ a diiron cofactor, here, we show that pAB synthesis by CADD requires manganese and the physiological cofactor is most likely a heterodinuclear Mn\/Fe cluster. Isotope\u2010labeling experiments revealed that the two oxygen atoms in the carboxylic acid portion of pAB are derived from molecular oxygen. Further, mass spectrometry\u2010based proteomic analyses of CADD\u2010derived peptides demonstrated a glycine substitution at Tyr27, providing strong evidence that this residue is sacrificed for pAB synthesis. Additionally, Lys152 was deaminated and oxidized to aminoadipic acid, supporting its proposed role as a sacrificial amino group donor.<\/jats:p>","DOI":"10.1002\/1873-3468.14573","type":"journal-article","created":{"date-parts":[[2023,1,17]],"date-time":"2023-01-17T11:16:52Z","timestamp":1673954212000},"page":"557-572","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["The <i>Chlamydia trachomatis p<\/i>\u2010aminobenzoate synthase <scp>CADD<\/scp> is a manganese\u2010dependent oxygenase that uses its own amino acid residues as substrates"],"prefix":"10.1002","volume":"597","author":[{"given":"Rowan","family":"Wooldridge","sequence":"first","affiliation":[{"name":"Department of Biochemistry Virginia Tech  Blacksburg VA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Spenser","family":"Stone","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Virginia Tech  Blacksburg VA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew","family":"Pedraza","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Virginia Tech  Blacksburg VA USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"W. 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