{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,24]],"date-time":"2025-10-24T07:55:20Z","timestamp":1761292520155},"reference-count":52,"publisher":"Portland Press Ltd.","issue":"3","content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2008,5,1]]},"abstract":"<jats:p>BopE is a type\u00a0III secreted protein from Burkholderia pseudomallei, the aetiological agent of melioidosis, a severe emerging infection. BopE is a GEF (guanine-nucleotide-exchange factor) for the Rho GTPases Cdc42 (cell division cycle 42) and Rac1. We have determined the structure of BopE catalytic domain (amino acids 78\u2013261) by NMR spectroscopy and it shows that BopE78\u2013261 comprises two three-helix bundles (\u03b11\u03b14\u03b15 and \u03b12\u03b13\u03b16). This fold is similar to that adopted by the BopE homologues SopE and SopE2, which are GEFs from Salmonella. Whereas the two three-helix bundles of SopE78\u2013240 and SopE269\u2013240 form the arms of a \u2018\u039b\u2019 shape, BopE78\u2013261 adopts a more closed conformation with substantial interactions between the two three-helix bundles. We propose that arginine and proline residues are important in the conformational differences between BopE and SopE\/E2. Analysis of the molecular interface in the SopE78\u2013240\u2013Cdc42 complex crystal structure indicates that, in a BopE\u2013Cdc42 interaction, the closed conformation of BopE78\u2013261 would engender steric clashes with the Cdc42 switch regions. This implies that BopE78\u2013261 must undergo a closed-to-open conformational change in order to catalyse guanine nucleotide exchange. In an NMR titration to investigate the BopE78\u2013261\u2013Cdc42 interaction, the appearance of additional peaks per NH for residues in hinge regions of BopE78\u2013261 indicates that BopE78\u2013261 does undergo a closed-to-open conformational change in the presence of Cdc42. The conformational change hypothesis is further supported by substantial improvement of BopE78\u2013261 catalytic efficiency through mutations that favour an open conformation. Requirement for closed-to-open conformational change explains the 10\u201340-fold lower kcat of BopE compared with SopE and SopE2.<\/jats:p>","DOI":"10.1042\/bj20071546","type":"journal-article","created":{"date-parts":[[2008,4,14]],"date-time":"2008-04-14T16:06:13Z","timestamp":1208189173000},"page":"485-493","update-policy":"http:\/\/dx.doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["The guanine-nucleotide-exchange factor BopE from <i>Burkholderia pseudomallei<\/i> adopts a compact version of the <i>Salmonella<\/i> SopE\/SopE2 fold and undergoes a closed-to-open conformational change upon interaction with Cdc42"],"prefix":"10.1042","volume":"411","author":[{"given":"Abhishek","family":"Upadhyay","sequence":"first","affiliation":[{"name":"Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Huan-Lin","family":"Wu","sequence":"additional","affiliation":[{"name":"Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher","family":"Williams","sequence":"additional","affiliation":[{"name":"Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Terry","family":"Field","sequence":"additional","affiliation":[{"name":"Division of Environmental Microbiology, IAH (Institute for Animal Health), Compton Laboratory, Berkshire RG20 7NN, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Edouard\u00a0E.","family":"Galyov","sequence":"additional","affiliation":[{"name":"Division of Environmental Microbiology, IAH (Institute for Animal Health), Compton Laboratory, Berkshire RG20 7NN, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jean\u00a0M.\u00a0H.","family":"van\u00a0den\u00a0Elsen","sequence":"additional","affiliation":[{"name":"Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Stefan","family":"Bagby","sequence":"additional","affiliation":[{"name":"Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, U.K."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"288","published-online":{"date-parts":[[2008,4,14]]},"reference":[{"key":"2021112215162136000_B1","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1097\/00001432-200204000-00005","article-title":"Melioidosis","volume":"15","author":"Dance","year":"2002","journal-title":"Curr. 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