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Here, the crystal structure of a relatively protease-resistant N-terminal fragment of human Cwc27 containing the PPIase domain was determined at 2.0\u2005\u00c5 resolution. The fragment exhibits a C-terminal appendix and resides in a reduced state compared with the previous oxidized structure of a similar fragment. By combining multiple sequence alignments spanning the eukaryotic tree of life and secondary-structure prediction, Cwc27 proteins across the entire eukaryotic kingdom were identified. This analysis revealed the specific loss of a crucial active-site residue in higher eukaryotic Cwc27 proteins, suggesting that the protein evolved from a prolyl isomerase to a pure proline binder. Noting a fungus-specific insertion in the PPIase domain, the 1.3\u2005\u00c5 resolution crystal structure of the PPIase domain of Cwc27 from<jats:italic>Chaetomium thermophilum<\/jats:italic>was also determined. Although structurally highly similar in the core domain, the<jats:italic>C. thermophilum<\/jats:italic>protein displayed a higher thermal stability than its human counterpart, presumably owing to the combined effect of several amino-acid exchanges that reduce the number of long side chains with strained conformations and create new intramolecular interactions, in particular increased hydrogen-bond networks.<\/jats:p>","DOI":"10.1107\/s1399004714021695","type":"journal-article","created":{"date-parts":[[2014,11,21]],"date-time":"2014-11-21T19:29:15Z","timestamp":1416598155000},"page":"3110-3123","update-policy":"https:\/\/doi.org\/10.1107\/cm_01","source":"Crossref","is-referenced-by-count":23,"title":["Structure and evolution of the spliceosomal peptidyl-prolyl<i>cis<\/i>\u2013<i>trans<\/i>isomerase Cwc27"],"prefix":"10.1107","volume":"70","author":[{"given":"Alexander","family":"Ulrich","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Markus C.","family":"Wahl","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"329","published-online":{"date-parts":[[2014,11,22]]},"reference":[{"key":"dw5118_bb1","doi-asserted-by":"crossref","first-page":"489","DOI":"10.1038\/nrm3392","volume":"13","author":"Adrain","year":"2012","journal-title":"Nature Rev. 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