{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,3]],"date-time":"2026-06-03T17:55:58Z","timestamp":1780509358760,"version":"3.54.1"},"reference-count":39,"publisher":"Wiley","issue":"16","license":[{"start":{"date-parts":[[2015,3,20]],"date-time":"2015-03-20T00:00:00Z","timestamp":1426809600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100004396","name":"Thailand Research Fund","doi-asserted-by":"crossref","award":["MRG5580066"],"award-info":[{"award-number":["MRG5580066"]}],"id":[{"id":"10.13039\/501100004396","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/501100004396","name":"Thailand Research Fund","doi-asserted-by":"crossref","award":["RTA5680001"],"award-info":[{"award-number":["RTA5680001"]}],"id":[{"id":"10.13039\/501100004396","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Faculty of Science, Burapha University, Thailand"},{"name":"Faculty of Science, Mahidol University, Thailand"}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FEBS Journal"],"published-print":{"date-parts":[[2015,8]]},"abstract":"<jats:p><jats:sc>l<\/jats:sc>\u2013\u03b1\u2010glycerophosphate oxidase is an <jats:styled-content style=\"fixed-case\">FAD<\/jats:styled-content>\u2010dependent enzyme that catalyzes the oxidation of <jats:sc>l<\/jats:sc>\u2013\u03b1\u2010glycerophosphate (Glp) by molecular oxygen to generate dihydroxyacetone phosphate (<jats:styled-content style=\"fixed-case\">DHAP<\/jats:styled-content>) and hydrogen peroxide (H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub>). The catalytic properties of recombinant His<jats:sub>6<\/jats:sub>\u2010GlpO from <jats:italic><jats:styled-content style=\"fixed-case\">M<\/jats:styled-content>ycoplasma\u00a0pneumoniae<\/jats:italic> (His<jats:sub>6<\/jats:sub>\u2010<jats:italic>Mp<\/jats:italic>GlpO) were investigated through transient and steady\u2010state kinetics and ligand binding studies. The results indicate that the reaction mechanism of His<jats:sub>6<\/jats:sub>\u2010<jats:italic>Mp<\/jats:italic>GlpO follows a ping\u2010pong model. Double\u2010mixing mode stopped\u2010flow experiments show that, after flavin\u2010mediated substrate oxidation, <jats:styled-content style=\"fixed-case\">DHAP<\/jats:styled-content> leaves rapidly prior to the oxygen reaction. The values determined for the individual rate constants and <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> (4.2\u00a0s<jats:sup>\u22121<\/jats:sup> at 4\u00a0\u00b0C), in addition to the finding that H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> binds to the oxidized enzyme, suggest that H<jats:sub>2<\/jats:sub>O<jats:sub>2<\/jats:sub> release is the rate\u2010limiting step for the overall reaction. The results indicate that His<jats:sub>6<\/jats:sub>\u2010<jats:italic>Mp<\/jats:italic>GlpO contains mixed populations of fast\u2010 and slow\u2010reacting species. It is predominantly the fast\u2010reacting species that participates in turnover. In contrast to other GlpO enzymes previously described, His<jats:sub>6<\/jats:sub>\u2010<jats:italic>Mp<\/jats:italic>GlpO is able to catalyze the reverse reaction of reduced enzyme and <jats:styled-content style=\"fixed-case\">DHAP<\/jats:styled-content>. This result may be explained by the standard reduction potential value of His<jats:sub>6<\/jats:sub>\u2010<jats:italic>Mp<\/jats:italic>GlpO (\u2212167\u00a0\u00b1\u00a01\u00a0mV), which is lower than those of GlpO from other species. We found that <jats:sc>d,l<\/jats:sc>\u2013glyceraldehyde 3\u2013phosphate (<jats:styled-content style=\"fixed-case\">GAP<\/jats:styled-content>) may be used as a substrate in the His<jats:sub>6<\/jats:sub>\u2010<jats:italic>Mp<\/jats:italic>GlpO reaction, although it exhibited an approximately 100\u2010fold lower <jats:italic>k<\/jats:italic><jats:sub>cat<\/jats:sub> value in comparison with the reaction of Glp. These results also imply involvement of GlpO in glycolysis, as well as in lipid and glycerol metabolism. The kinetic models and distinctive properties of His<jats:sub>6<\/jats:sub>\u2010<jats:italic>Mp<\/jats:italic>GlpO reported here should be useful for future drug development against <jats:italic>Mycoplasma\u00a0pneumoniae<\/jats:italic> infection.<\/jats:p>","DOI":"10.1111\/febs.13247","type":"journal-article","created":{"date-parts":[[2015,2,25]],"date-time":"2015-02-25T08:19:12Z","timestamp":1424852352000},"page":"3043-3059","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":23,"title":["Kinetic mechanism of <scp>l<\/scp>\u2013\u03b1\u2013glycerophosphate oxidase from <i>Mycoplasma\u00a0pneumoniae<\/i>"],"prefix":"10.1111","volume":"282","author":[{"given":"Somchart","family":"Maenpuen","sequence":"first","affiliation":[{"name":"Department of Biochemistry Faculty of Science Burapha University Chonburi Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pratchaya","family":"Watthaisong","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Faculty of Science Burapha University Chonburi Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pacharee","family":"Supon","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Faculty of Science Burapha University Chonburi Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeerus","family":"Sucharitakul","sequence":"additional","affiliation":[{"name":"Department of Biochemistry Faculty of Dentistry Chulalongkorn University Patumwan Bangkok Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Derek","family":"Parsonage","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Center for Structural Biology Wake Forest School of Medicine Winston\u2010Salem NC USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"P. Andrew","family":"Karplus","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Biophysics Oregon State University Corvallis OR USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Al","family":"Claiborne","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Center for Structural Biology Wake Forest School of Medicine Winston\u2010Salem NC USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pimchai","family":"Chaiyen","sequence":"additional","affiliation":[{"name":"Department of Biochemistry and Center of Excellence in Protein Structure &amp; Function Faculty of Science Mahidol University Bangkok Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2015,3,20]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1128\/jb.98.3.1063-1068.1969"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(17)30130-8"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)66883-8"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1159\/000096470"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.01103-08"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2007.03.016"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI111279"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1128\/JB.187.19.6824-6831.2005"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.17.4.697-728.2004"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-4894(77)90040-6"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/0014-5793(76)80045-2"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/0020-711X(77)90104-5"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/0020-711X(77)90105-7"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/0166-6851(86)90079-4"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.273.37.23812"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi992499j"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444901018169"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi701685u"},{"key":"e_1_2_8_20_1","first-page":"132","volume-title":"Structure and Mechanism in Protein Science: A Guide to Enzyme Catalysis and Folding","author":"Fersht A","year":"1999"},{"key":"e_1_2_8_21_1","volume-title":"Data for Biochemical Research","author":"Dawson RMC","year":"1986"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi035349o"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1128\/CDLI.8.1.85-92.2001"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1111\/febs.13233"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi801039d"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1021\/ja4088055"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M110.163881"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M111.241836"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi800228w"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0903809106"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2012.06.005"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi802331r"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.073247"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(76)90527-3"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.1385\/1-59259-266-X:1"},{"key":"e_1_2_8_36_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1993.tb17982.x"},{"key":"e_1_2_8_37_1","doi-asserted-by":"publisher","DOI":"10.3891\/acta.chem.scand.11-1706"},{"key":"e_1_2_8_38_1","doi-asserted-by":"publisher","DOI":"10.1021\/ed1000125"},{"key":"e_1_2_8_39_1","first-page":"59","volume-title":"Flavins and Flavoproteins","author":"Massey V","year":"1991"},{"key":"e_1_2_8_40_1","volume-title":"Compilations of Data in Oxidation\u2010Reduction Potentials of Organic Systems","author":"Clark WM","year":"1960"}],"container-title":["The FEBS Journal"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ffebs.13247","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Ffebs.13247","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/febs.13247","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,13]],"date-time":"2023-09-13T12:07:44Z","timestamp":1694606864000},"score":1,"resource":{"primary":{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/10.1111\/febs.13247"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2015,3,20]]},"references-count":39,"journal-issue":{"issue":"16","published-print":{"date-parts":[[2015,8]]}},"alternative-id":["10.1111\/febs.13247"],"URL":"https:\/\/doi.org\/10.1111\/febs.13247","archive":["Portico"],"relation":{},"ISSN":["1742-464X","1742-4658"],"issn-type":[{"value":"1742-464X","type":"print"},{"value":"1742-4658","type":"electronic"}],"subject":[],"published":{"date-parts":[[2015,3,20]]},"assertion":[{"value":"2014-11-27","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-02-20","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2015-03-20","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}