{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T05:14:59Z","timestamp":1780377299387,"version":"3.54.1"},"reference-count":28,"publisher":"Wiley","issue":"11","license":[{"start":{"date-parts":[[2020,9,17]],"date-time":"2020-09-17T00:00:00Z","timestamp":1600300800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Protein Science"],"published-print":{"date-parts":[[2020,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:sc>l<\/jats:sc>\u2010Lysine oxidase (LysOX) is a FAD\u2010dependent homodimeric enzyme that catalyzes the oxidative deamination of <jats:sc>l<\/jats:sc>\u2010lysine to produce \u03b1\u2010keto\u2010\u03b5\u2010aminocaproate with ammonia and hydrogen peroxide. LysOX shows strict substrate specificity for <jats:sc>l<\/jats:sc>\u2010lysine, whereas most <jats:sc>l<\/jats:sc>\u2010amino acid oxidases (LAAOs) exhibit broad substrate specificity for <jats:sc>l<\/jats:sc>\u2010amino acids. Previous studies of LysOX showed that overall structural similarity to the well\u2010studied snake venom LAAOs. However, the molecular mechanism of strict specificity for <jats:sc>l<\/jats:sc>\u2010lysine was still unclear. We here determined the structure of LysOX in complex with <jats:sc>l<\/jats:sc>\u2010lysine at 1.7 \u00c5 resolution. The structure revealed that the hydrogen bonding network formed by D212, D315, and A440 with two water molecules is responsible for the recognition of the side chain amino group. In addition, a narrow hole formed by five hydrophobic residues in the active site contributes to strict substrate specificity. Mutation studies demonstrated that D212 and D315 are essential for <jats:sc>l<\/jats:sc>\u2010lysine recognition, and the D212A\/D315A double mutant LysOX showed different substrate specificity from LysOX. Moreover, the structural basis of the substrate specificity change has also been revealed by the structural analysis of the mutant variant and its substrate complexes. These results clearly explain the molecular mechanism of the strict specificity of LysOX and suggest that LysOX is a potential candidate for a template to design LAAOs specific to other <jats:sc>l<\/jats:sc>\u2010amino acids.<\/jats:p>","DOI":"10.1002\/pro.3946","type":"journal-article","created":{"date-parts":[[2020,9,11]],"date-time":"2020-09-11T08:04:05Z","timestamp":1599811445000},"page":"2213-2225","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":12,"title":["Structural basis of strict substrate recognition of <scp>l<\/scp>\u2010lysine \u03b1\u2010oxidase from <scp><i>Trichoderma viride<\/i><\/scp>"],"prefix":"10.1002","volume":"29","author":[{"given":"Hiroki","family":"Kondo","sequence":"first","affiliation":[{"name":"Department of Macromolecular Science, Graduate School of Science Osaka University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaki","family":"Kitagawa","sequence":"additional","affiliation":[{"name":"Department of Macromolecular Science, Graduate School of Science Osaka University  Osaka Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuya","family":"Matsumoto","sequence":"additional","affiliation":[{"name":"Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science Okayama University  Okayama Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Masaya","family":"Saito","sequence":"additional","affiliation":[{"name":"Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science Okayama University  Okayama Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Amano","sequence":"additional","affiliation":[{"name":"Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science Okayama University  Okayama Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shigeru","family":"Sugiyama","sequence":"additional","affiliation":[{"name":"Faculty of Science and Technology Kochi University  Kochi Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Takashi","family":"Tamura","sequence":"additional","affiliation":[{"name":"Department of Biofunctional Chemistry, Graduate School of Environmental and Life Science Okayama University  Okayama Japan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hitoshi","family":"Kusakabe","sequence":"additional","affiliation":[{"name":"Enzyme Sensor Co., Ltd.  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