{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T01:13:00Z","timestamp":1790817180597,"version":"4.1.0"},"reference-count":59,"publisher":"Wiley","issue":"9","license":[{"start":{"date-parts":[[2019,6,21]],"date-time":"2019-06-21T00:00:00Z","timestamp":1561075200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003762","name":"Institut Pasteur","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100003762","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["faseb.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The FASEB Journal"],"published-print":{"date-parts":[[2019,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    The adenylate cyclase (CyaA) toxin is a major virulence factor of\n                    <jats:italic>Bordetella pertussis<\/jats:italic>\n                    , the causative agent of whooping cough. CyaA is synthetized as a pro\u2010toxin, pro\u2010CyaA, and converted into its cytotoxic form upon acylation of two lysines. After secretion, CyaA invades eukaryotic cells and produces cAMP, leading to host defense subversion. To gain further insights into the effect of acylation, we compared the functional and structural properties of pro\u2010CyaA and CyaA proteins. HDX\u2010MS results show that the refolding process of both proteins upon progressive urea removal is initiated by calcium binding to the C\u2010terminal RTX domain. We further identified a critical hydrophobic segment, distal from the acylation region, that folds at higher urea concentration in CyaA than in pro\u2010CyaA. Once refolded into monomers, CyaA is more compact and stable than pro\u2010CyaA, due to a complex set of interactions between domains. Our HDX\u2010MS data provide direct evidence that the presence of acyl chains in CyaA induces a significant stabilization of the apolar segments of the hydrophobic domain and of most of the acylation region. We propose a refolding model dependent on calcium and driven by local and distal acylation\u2010dependent interactions within CyaA. Therefore, CyaA acylation is not only critical for cell intoxication, but also for protein refolding into its active conformation. Our data shed light on the complex relationship between post\u2010translational modifications, structural disorder and protein folding. Coupling calcium\u2010binding and acylation\u2010driven folding is likely pertinent for other repeat\u2010in\u2010toxin cytolysins produced by many Gram\u2010negative bacterial pathogens.\u2014O'Brien, D. P., Cannella, S. E., Voegele, A., Raoux\u2010Barbot, D., Davi, M., Douch\u00e9, T., Matondo, M., Brier, S., Ladant, D., Chenal, A. Post\u2010translational acylation controls the folding and functions of the CyaA RTX toxin. FASEB J. 33, 10065\u201310076 (2019).\n                    <jats:ext-link xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"http:\/\/www.fasebj.org\">www.fasebj.org<\/jats:ext-link>\n                  <\/jats:p>","DOI":"10.1096\/fj.201802442rr","type":"journal-article","created":{"date-parts":[[2019,6,21]],"date-time":"2019-06-21T15:42:11Z","timestamp":1561131731000},"page":"10065-10076","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":32,"title":["Post\u2010translational acylation controls the folding and functions of the CyaA RTX toxin"],"prefix":"10.1096","volume":"33","author":[{"given":"Darragh P.","family":"O'Brien","sequence":"first","affiliation":[{"name":"Institut Pasteur Chemistry and Structural Biology Department  Paris France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sara E.","family":"Cannella","sequence":"additional","affiliation":[{"name":"Institut Pasteur Chemistry and 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