{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,18]],"date-time":"2025-10-18T10:14:55Z","timestamp":1760782495200},"reference-count":0,"publisher":"Wiley","issue":"3","license":[{"start":{"date-parts":[[2001,12,25]],"date-time":"2001-12-25T00:00:00Z","timestamp":1009238400000},"content-version":"vor","delay-in-days":1423,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European Journal of Biochemistry"],"published-print":{"date-parts":[[1998,2]]},"abstract":"<jats:p>The <jats:italic>Leishmania<\/jats:italic> short\u2010chain dehydrogenase\/reductase gene <jats:italic>ptr1<\/jats:italic> has been isolated while characterizing antifolate\u2010resistant <jats:italic>Leishmania<\/jats:italic> mutants. PTR1 is active as a tetramer and can reduce pterins and folates. PTR1 has several of the hallmarks of the short chain dehydrogenase\/reductase family, including a glycine rich co\u2010factor binding site at its N\u2010terminus, and the consensus catalytic site TyrXaa<jats:sub>3<\/jats:sub>Lys. To start probing the structure\/function of PTR1, we have generated by site\u2010directed mutagenesis five mutants, either in the co\u2010factor\u2212binding site, Y38D or in the catalytic site Y195F, Y195W, K199R or in a PTR1\u2010specific region, Y175F. The mutated versions of PTR1 were studied <jats:italic>in vivo<\/jats:italic> in <jats:italic>Leishmania<\/jats:italic> and at the biochemical level using purified proteins. The Y175F mutant showed properties similar to wild\u2010type PTR1 in every aspect tested, but all the other mutants were inactive even if they were purified as tetramers. To test the ability of the mutated PTR1 versions to bind its co\u2010factor and substrates, trypsin digestion experiments were carried out under conditions upon which binding will prevent wild\u2010type PTR1 of being digested by trypsin. The wild\u2010type PTR1 as well as Y175F and Y195F mutants were protected against trypsin digestion whereas Y38D and K199R mutants were not. Mutations in regions involved in co\u2010factor binding (Y38D) or in catalytic site (K199R) altered the binding of the ligands, explaining why those protein versions are inactive.<\/jats:p>","DOI":"10.1046\/j.1432-1327.1998.2510768.x","type":"journal-article","created":{"date-parts":[[2003,3,11]],"date-time":"2003-03-11T16:51:26Z","timestamp":1047401486000},"page":"768-774","source":"Crossref","is-referenced-by-count":15,"title":["Residues involved in co\u2010factor and substrate binding of the short\u2010chain dehydrogenase\/reductase PTR1 producing methotrexate resistance in <i>Leishmania<\/i>"],"prefix":"10.1111","volume":"251","author":[{"given":"\u00c9ric","family":"Leblanc","sequence":"first","affiliation":[]},{"given":"Barbara","family":"Papadopoulou","sequence":"additional","affiliation":[]},{"given":"Chantale","family":"Bernatchez","sequence":"additional","affiliation":[]},{"given":"Marc","family":"Ouellette","sequence":"additional","affiliation":[]}],"member":"311","published-online":{"date-parts":[[2001,12,25]]},"container-title":["European Journal of Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1046%2Fj.1432-1327.1998.2510768.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/pdf\/10.1046\/j.1432-1327.1998.2510768.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,23]],"date-time":"2023-11-23T14:24:07Z","timestamp":1700749447000},"score":1,"resource":{"primary":{"URL":"https:\/\/febs.onlinelibrary.wiley.com\/doi\/10.1046\/j.1432-1327.1998.2510768.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1998,2]]},"references-count":0,"journal-issue":{"issue":"3","published-print":{"date-parts":[[1998,2]]}},"alternative-id":["10.1046\/j.1432-1327.1998.2510768.x"],"URL":"https:\/\/doi.org\/10.1046\/j.1432-1327.1998.2510768.x","archive":["Portico"],"relation":{},"ISSN":["0014-2956","1432-1033"],"issn-type":[{"value":"0014-2956","type":"print"},{"value":"1432-1033","type":"electronic"}],"subject":[],"published":{"date-parts":[[1998,2]]}}}