{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,9]],"date-time":"2026-10-09T20:04:43Z","timestamp":1791576283154,"version":"4.3.4"},"reference-count":68,"publisher":"Wiley","issue":"52","license":[{"start":{"date-parts":[[2017,8,21]],"date-time":"2017-08-21T00:00:00Z","timestamp":1503273600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Chemistry A European J"],"published-print":{"date-parts":[[2017,9,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Isopenicillin N synthase (IPNS) catalyses the four\u2010electron oxidation of a tripeptide,\n                    <jats:sc>l<\/jats:sc>\n                    \u2010\u03b4\u2010(\u03b1\u2010aminoadipoyl)\u2010\n                    <jats:sc>l<\/jats:sc>\n                    \u2010cysteinyl\u2010\n                    <jats:sc>d<\/jats:sc>\n                    \u2010valine (ACV), to give isopenicillin N (IPN), the first\u2010formed \u03b2\u2010lactam in penicillin and cephalosporin biosynthesis. IPNS catalysis is dependent upon an iron(II) cofactor and oxygen as a co\u2010substrate. In the absence of substrate, the carbonyl oxygen of the side\u2010chain amide of the penultimate residue, Gln330, co\u2010ordinates to the active\u2010site metal iron. Substrate binding ablates the interaction between Gln330 and the metal, triggering rearrangement of seven C\u2010terminal residues, which move to take up a conformation that extends the final \u03b1\u2010helix and encloses ACV in the active site. Mutagenesis studies are reported, which probe the role of the C\u2010terminal and other aspects of the substrate binding pocket in IPNS. The hydrophobic nature of amino acid side\u2010chains around the ACV binding pocket is important in catalysis. Deletion of seven C\u2010terminal residues exposes the active site and leads to formation of a new type of thiol oxidation product. The isolated product is shown by LC\u2010MS and NMR analyses to be the ene\u2010thiol tautomer of a dithioester, made up from two molecules of ACV linked between the thiol sulfur of one tripeptide and the oxidised cysteinyl \u03b2\u2010carbon of the other. A mechanism for its formation is proposed, supported by an X\u2010ray crystal structure, which shows the substrate ACV bound at the active site, its cysteinyl \u03b2\u2010carbon exposed to attack by a second molecule of substrate, adjacent. Formation of this product constitutes a new mode of reaction for IPNS and non\u2010heme iron oxidases in general.\n                  <\/jats:p>","DOI":"10.1002\/chem.201701592","type":"journal-article","created":{"date-parts":[[2017,7,13]],"date-time":"2017-07-13T06:11:59Z","timestamp":1499926319000},"page":"12815-12824","source":"Crossref","is-referenced-by-count":16,"title":["Terminally Truncated Isopenicillin N Synthase Generates a Dithioester Product: Evidence for a Thioaldehyde Intermediate during Catalysis and a New Mode of Reaction for Non\u2010Heme Iron Oxidases"],"prefix":"10.1002","volume":"23","author":[{"given":"Luke A.","family":"McNeill","sequence":"first","affiliation":[{"name":"Oxford Centre for Molecular Sciences and the Department of Chemistry Chemistry Research Laboratory  Mansfield Road Oxford OX1 3TA UK"},{"name":"Present Address: Oxford Nanopore Technologies, Oxford Science Park  OX4 4GA UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Toby J. N.","family":"Brown","sequence":"additional","affiliation":[{"name":"Oxford Centre for Molecular Sciences and the Department of Chemistry Chemistry Research Laboratory  Mansfield Road Oxford OX1 3TA UK"},{"name":"Present Address: The Brattle Group  Level 15 5 Martin Place Sydney, NSW 2000 Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Malkit","family":"Sami","sequence":"additional","affiliation":[{"name":"Oxford Centre for Molecular Sciences and the Department of Chemistry Chemistry Research Laboratory  Mansfield Road Oxford OX1 3TA UK"},{"name":"Present Address: Immunocore Limited  101 Park Drive, Milton Park Abingdon OX14 4RY UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ian J.","family":"Clifton","sequence":"additional","affiliation":[{"name":"Oxford Centre for Molecular Sciences and the Department of Chemistry Chemistry Research Laboratory  Mansfield Road Oxford OX1 3TA UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolai I.","family":"Burzlaff","sequence":"additional","affiliation":[{"name":"Department of Chemistry and Pharmacy University of Erlangen-Nuremberg  Egerlandstra\u00dfe 1 91058 Erlangen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Timothy D. 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