{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,19]],"date-time":"2025-10-19T18:24:04Z","timestamp":1760898244305},"reference-count":41,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2013,12,5]],"date-time":"2013-12-05T00:00:00Z","timestamp":1386201600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/3.0\/"}],"content-domain":{"domain":["febs.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["FEBS Open Bio"],"published-print":{"date-parts":[[2014,1]]},"abstract":"<jats:p>Centrins are members of the EF\u2010hand family of calcium\u2010binding proteins, which are highly conserved among eukaryotes. Centrins bind to several cellular targets, through a hydrophobic triad. However, the W<jats:sup>1<\/jats:sup>xxL<jats:sup>4<\/jats:sup>xxxL<jats:sup>8<\/jats:sup> triad in XPC (Xeroderma Pigmentosum Group C protein) is found in the reverse orientation, as in the L<jats:sup>8<\/jats:sup>xxxL<jats:sup>4<\/jats:sup>xxW<jats:sup>1<\/jats:sup> triad in Sfi1 (Suppressor of Fermentation\u2010Induced loss of stress resistance protein 1). As shown by previous NMR studies of human centrin 2 in complex with XPC or Sfi1, the E148 residue of human centrin 2 is in contact with XPC but is pushed away from the triad of Sfi1. We corroborated these findings using site\u2010directed mutagenesis to generate mutations in <jats:italic>Scherffelia dubia<\/jats:italic> centrin (SdCen) and by using isothermal titration calorimetry to analyze the binding affinity of these mutants to XPC and Sfi1. We mutated the F109 residue, which is the main residue involved in target binding regardless of triad orientation, and the E144 residue, which was thought to be involved only in XPC binding. The F109L mutation reduced the binding of SdCen to XPC and Sfi1 and the negative effect was greater upon temperature increase. By contrast, the E144A mutation reduced the binding to XPC but had no effect on Sfi1 binding. The F109L\u2010E144A mutation enhanced the negative effect of the two single mutations on XPC binding. Sfi1 proteins from <jats:italic>Ostreococcus lucimarinus<\/jats:italic> and <jats:italic>Ostreococcus tauri<\/jats:italic>, which belong to the same clade as <jats:italic>S. dubia,<\/jats:italic> were also investigated. A comparative analysis shows that the triad residues are more conserved than those in human Sfi1.<\/jats:p>","DOI":"10.1016\/j.fob.2013.11.005","type":"journal-article","created":{"date-parts":[[2013,12,5]],"date-time":"2013-12-05T21:32:06Z","timestamp":1386279126000},"page":"33-42","update-policy":"http:\/\/dx.doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The E144 residue of <i>Scherffelia dubia<\/i> centrin discriminates between the DNA repair protein XPC and the centrosomal protein Sfi1"],"prefix":"10.1002","volume":"4","author":[{"given":"Dora","family":"Grecu","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yves","family":"Blouquit","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liliane","family":"Assairi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2013,12,5]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.99.3.962"},{"key":"e_1_2_8_3_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.107.1.133"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.124.5.795"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1104\/pp.99.4.1734"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.83.15.5512"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/0248-4900(92)90441-3"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.23.11039"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.107.1.9"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1996.0121q.x"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.37.22453"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1093\/genetics\/157.2.503"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E02-10-0661"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1242\/jcs.00497"},{"key":"e_1_2_8_15_1","doi-asserted-by":"publisher","DOI":"10.1105\/tpc.021378"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M100855200"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1038\/ncb1163"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.22.7.2194-2203.2002"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M406770200"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200307064"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1002\/(SICI)1097-0061(199908)15:11<1097::AID-YEA437>3.0.CO;2-B"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M509886200"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi901764m"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi051397s"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M513667200"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2007.08.046"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi0524868"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M101324200"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2009.01.033"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200603153"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmb.2009.10.041"},{"key":"e_1_2_8_32_1","first-page":"183","article-title":"Rapid and efficient one-tube PCR-based mutagenesis method","volume":"67","author":"Picard V.","year":"1997","journal-title":"Methods Mol. 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