{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,1]],"date-time":"2026-10-01T01:13:01Z","timestamp":1790817181249,"version":"4.1.0"},"reference-count":80,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,2,10]],"date-time":"2017-02-10T00:00:00Z","timestamp":1486684800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,2,10]],"date-time":"2017-02-10T00:00:00Z","timestamp":1486684800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    <jats:italic>Bordetella pertussis<\/jats:italic>\n                    , the causative agent of whooping cough, secretes an adenylate cyclase toxin, CyaA, which invades eukaryotic cells and alters their physiology by cAMP overproduction. Calcium is an essential cofactor of CyaA, as it is the case for most members of the Repeat-in-ToXins (RTX) family. We show that the calcium-bound, monomeric form of CyaA, hCyaAm, conserves its permeabilization and haemolytic activities, even in a fully calcium-free environment. In contrast, hCyaAm requires sub-millimolar calcium in solution for cell invasion, indicating that free calcium in solution is involved in the CyaA toxin translocation process. We further report the first in solution structural characterization of hCyaAm, as deduced from SAXS, mass spectrometry and hydrodynamic studies. We show that hCyaAm adopts a compact and stable state that can transiently conserve its conformation even in a fully calcium-free environment. Our results therefore suggest that in hCyaAm, the C-terminal RTX-domain is stabilized in a high-affinity calcium-binding state by the N-terminal domains while, conversely, calcium binding to the C-terminal RTX-domain strongly stabilizes the N-terminal regions. Hence, the different regions of hCyaAm appear tightly connected, leading to stabilization effects between domains. The hysteretic behaviour of CyaA in response to calcium is likely shared by other RTX cytolysins.\n                  <\/jats:p>","DOI":"10.1038\/srep42065","type":"journal-article","created":{"date-parts":[[2017,2,10]],"date-time":"2017-02-10T05:15:13Z","timestamp":1486703713000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":46,"title":["Stability, structural and functional properties of a monomeric, calcium\u2013loaded adenylate cyclase toxin, CyaA, from Bordetella pertussis"],"prefix":"10.1038","volume":"7","author":[{"given":"Sara E.","family":"Cannella","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V\u00e9ronique Yvette","family":"Ntsogo Engu\u00e9n\u00e9","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marilyne","family":"Davi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Christian","family":"Malosse","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Cristina","family":"Sotomayor P\u00e9rez","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia","family":"Chamot-Rooke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrice","family":"Vachette","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominique","family":"Durand","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Ladant","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexandre","family":"Chenal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,2,10]]},"reference":[{"key":"BFsrep42065_CR1","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1016\/S0966-842X(99)01468-7","volume":"7","author":"D Ladant","year":"1999","unstructured":"Ladant, D. & Ullmann, A. Bordatella pertussis adenylate cyclase: a toxin with multiple talents. Trends in microbiology 7, 172\u2013176 (1999).","journal-title":"Trends in microbiology"},{"key":"BFsrep42065_CR2","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.mib.2005.12.011","volume":"9","author":"J Vojtova","year":"2006","unstructured":"Vojtova, J., Kamanova, J. & Sebo, P. Bordetella adenylate cyclase toxin: a swift saboteur of host defense. Curr Opin Microbiol 9, 69\u201375 (2006).","journal-title":"Curr Opin Microbiol"},{"key":"BFsrep42065_CR3","doi-asserted-by":"publisher","first-page":"88","DOI":"10.1016\/j.mib.2009.01.001","volume":"12","author":"R Shrivastava","year":"2009","unstructured":"Shrivastava, R. & Miller, J. F. Virulence factor secretion and translocation by Bordetella species. Curr Opin Microbiol 12, 88\u201393, doi: 10.1016\/j.mib.2009.01.001 (2009).","journal-title":"Curr Opin Microbiol"},{"key":"BFsrep42065_CR4","doi-asserted-by":"publisher","first-page":"455","DOI":"10.2217\/fmb.09.133","volume":"5","author":"NH Carbonetti","year":"2010","unstructured":"Carbonetti, N. H. Pertussis toxin and adenylate cyclase toxin: key virulence factors of Bordetella pertussis and cell biology tools. Future Microbiol 5, 455\u2013469, doi: 10.2217\/fmb.09.133 (2010).","journal-title":"Future Microbiol"},{"key":"BFsrep42065_CR5","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1111\/j.1365-2958.1988.tb00003.x","volume":"2","author":"P Glaser","year":"1988","unstructured":"Glaser, P. et al. The calmodulin-sensitive adenylate cyclase of Bordetella pertussis: cloning and expression in Escherichia coli . Molecular microbiology 2, 19\u201330 (1988).","journal-title":"Molecular microbiology"},{"key":"BFsrep42065_CR6","doi-asserted-by":"publisher","first-page":"318","DOI":"10.1021\/bi9016389","volume":"49","author":"JC Karst","year":"2010","unstructured":"Karst, J. C. et al. Calmodulin-induced conformational and hydrodynamic changes in the catalytic domain of Bordetella pertussis adenylate cyclase toxin. Biochemistry 49, 318\u2013328, doi: 10.1021\/bi9016389 (2010).","journal-title":"Biochemistry"},{"key":"BFsrep42065_CR7","doi-asserted-by":"publisher","first-page":"21131","DOI":"10.1074\/jbc.M113.530410","volume":"289","author":"E Selwa","year":"2014","unstructured":"Selwa, E. et al. Allosteric Activation of Bordetella pertussis Adenylyl Cyclase by Calmodulin: Molecular Dynamics and Mutagenesis Studies. J Biol Chem 289, 21131\u201321141, doi: 10.1074\/jbc.M113.530410 (2014).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR8","doi-asserted-by":"crossref","first-page":"3997","DOI":"10.1002\/j.1460-2075.1988.tb03288.x","volume":"7","author":"P Glaser","year":"1988","unstructured":"Glaser, P., Sakamoto, H., Bellalou, J., Ullmann, A. & Danchin, A. Secretion of cyclolysin, the calmodulin-sensitive adenylate cyclase-haemolysin bifunctional protein of Bordetella pertussis. Embo J 7, 3997\u20134004 (1988).","journal-title":"Embo J"},{"key":"BFsrep42065_CR9","doi-asserted-by":"crossref","first-page":"3242","DOI":"10.1128\/iai.58.10.3242-3247.1990","volume":"58","author":"J Bellalou","year":"1990","unstructured":"Bellalou, J., Sakamoto, H., Ladant, D., Geoffroy, C. & Ullmann, A. Deletions affecting hemolytic and toxin activities of Bordetella pertussis adenylate cyclase. Infection and immunity 58, 3242\u20133247 (1990).","journal-title":"Infection and immunity"},{"key":"BFsrep42065_CR10","doi-asserted-by":"crossref","first-page":"13598","DOI":"10.1016\/S0021-9258(18)42254-5","volume":"267","author":"H Sakamoto","year":"1992","unstructured":"Sakamoto, H., Bellalou, J., Sebo, P. & Ladant, D. Bordetella pertussis adenylate cyclase toxin. Structural and functional independence of the catalytic and hemolytic activities. J Biol Chem 267, 13598\u201313602 (1992).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR11","doi-asserted-by":"publisher","first-page":"20473","DOI":"10.1073\/pnas.1312975110","volume":"110","author":"R Veneziano","year":"2013","unstructured":"Veneziano, R. et al. Bordetella pertussis adenylate cyclase toxin translocation across a tethered lipid bilayer. Proc Natl Acad Sci USA 110, 20473\u201320478, doi: 10.1073\/pnas.1312975110 (2013).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep42065_CR12","doi-asserted-by":"publisher","first-page":"9200","DOI":"10.1074\/jbc.M111.316166","volume":"287","author":"JC Karst","year":"2012","unstructured":"Karst, J. C. et al. Identification of a region that assists membrane insertion and translocation of the catalytic domain of Bordetella pertussis CyaA toxin. J Biol Chem 287, 9200\u20139212, doi: 10.1074\/jbc.M111.316166 (2012).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR13","doi-asserted-by":"publisher","first-page":"29137","DOI":"10.1038\/srep29137","volume":"6","author":"J Masin","year":"2016","unstructured":"Masin, J. et al. Negatively charged residues of the segment linking the enzyme and cytolysin moieties restrict the membrane-permeabilizing capacity of adenylate cyclase toxin. Sci Rep 6, 29137, doi: 10.1038\/srep29137 (2016).","journal-title":"Sci Rep"},{"key":"BFsrep42065_CR14","doi-asserted-by":"publisher","first-page":"32585","DOI":"10.1074\/jbc.M113.508838","volume":"288","author":"O Subrini","year":"2013","unstructured":"Subrini, O. et al. Characterization of a membrane-active peptide from the Bordetella pertussis CyaA toxin. J Biol Chem 288, 32585\u201332598, doi: 10.1074\/jbc.M113.508838 (2013).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR15","doi-asserted-by":"crossref","first-page":"720","DOI":"10.1128\/jb.173.2.720-726.1991","volume":"173","author":"EM Barry","year":"1991","unstructured":"Barry, E. M. et al. Bordetella pertussis adenylate cyclase toxin and hemolytic activities require a second gene, cyaC, for activation. J Bacteriol 173, 720\u2013726 (1991).","journal-title":"J Bacteriol"},{"key":"BFsrep42065_CR16","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/0378-1119(91)90459-O","volume":"104","author":"P Sebo","year":"1991","unstructured":"Sebo, P., Glaser, P., Sakamoto, H. & Ullmann, A. High-level synthesis of active adenylate cyclase toxin of Bordetella pertussis in a reconstructed Escherichia coli system. Gene 104, 19\u201324, doi: 0378-1119(91)90459-O (1991).","journal-title":"Gene"},{"key":"BFsrep42065_CR17","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1126\/science.7939682","volume":"266","author":"M Hackett","year":"1994","unstructured":"Hackett, M., Guo, L., Shabanowitz, J., Hunt, D. F. & Hewlett, E. L. Internal lysine palmitoylation in adenylate cyclase toxin from Bordetella pertussis. Science 266, 433\u2013435 (1994).","journal-title":"Science"},{"key":"BFsrep42065_CR18","doi-asserted-by":"crossref","first-page":"91","DOI":"10.1016\/S0378-1119(96)00412-X","volume":"180","author":"GD Westrop","year":"1996","unstructured":"Westrop, G. D., Hormozi, E. K., Da Costa, N. A., Parton, R. & Coote, J. G. Bordetella pertussis adenylate cyclase toxin: proCyaA and CyaC proteins synthesised separately in Escherichia coli produce active toxin in vitro . Gene 180, 91\u201399 (1996).","journal-title":"Gene"},{"key":"BFsrep42065_CR19","doi-asserted-by":"crossref","first-page":"12759","DOI":"10.1021\/bi050459b","volume":"44","author":"J Masin","year":"2005","unstructured":"Masin, J. et al. Acylation of lysine 860 allows tight binding and cytotoxicity of Bordetella adenylate cyclase on CD11b-expressing cells. Biochemistry 44, 12759\u201312766 (2005).","journal-title":"Biochemistry"},{"key":"BFsrep42065_CR20","doi-asserted-by":"publisher","first-page":"30702","DOI":"10.1074\/jbc.M114.580852","volume":"289","author":"JC Karst","year":"2014","unstructured":"Karst, J. C. et al. Calcium, Acylation, and Molecular Confinement Favor Folding of Bordetella pertussis Adenylate Cyclase CyaA Toxin into a Monomeric and Cytotoxic Form. J Biol Chem 289, 30702\u201330716, doi: 10.1074\/jbc.M114.580852 (2014).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR21","doi-asserted-by":"crossref","first-page":"38514","DOI":"10.1074\/jbc.M304387200","volume":"278","author":"M El-Azami-El-Idrissi","year":"2003","unstructured":"El-Azami-El-Idrissi, M. et al. Interaction of Bordetella pertussis adenylate cyclase with CD11b\/CD18: Role of toxin acylation and identification of the main integrin interaction domain. J Biol Chem 278, 38514\u201338521 (2003).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR22","first-page":"85","volume":"257","author":"RA Welch","year":"2001","unstructured":"Welch, R. A. RTX toxin structure and function: a story of numerous anomalies and few analogies in toxin biology. Current topics in microbiology and immunology 257, 85\u2013111 (2001).","journal-title":"Current topics in microbiology and immunology"},{"key":"BFsrep42065_CR23","doi-asserted-by":"publisher","first-page":"1076","DOI":"10.1111\/j.1574-6976.2010.00231.x","volume":"34","author":"I Linhartova","year":"2010","unstructured":"Linhartova, I. et al. RTX proteins: a highly diverse family secreted by a common mechanism. FEMS Microbiol Rev 34, 1076\u20131112, doi: 10.1111\/j.1574-6976.2010.00231.x (2010).","journal-title":"FEMS Microbiol Rev"},{"key":"BFsrep42065_CR24","doi-asserted-by":"crossref","first-page":"10686","DOI":"10.1016\/S0021-9258(18)87001-6","volume":"265","author":"F Gentile","year":"1990","unstructured":"Gentile, F., Knipling, L. G., Sackett, D. L. & Wolff, J. Invasive adenylyl cyclase of Bordetella pertussis. Physical, catalytic, and toxic properties. J Biol Chem 265, 10686\u201310692 (1990).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR25","doi-asserted-by":"crossref","first-page":"17503","DOI":"10.1016\/S0021-9258(19)47400-0","volume":"266","author":"EL Hewlett","year":"1991","unstructured":"Hewlett, E. L. et al. Adenylate cyclase toxin from Bordetella pertussis. Conformational change associated with toxin activity. J Biol Chem 266, 17503\u201317508 (1991).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR26","doi-asserted-by":"crossref","first-page":"26370","DOI":"10.1074\/jbc.270.44.26370","volume":"270","author":"T Rose","year":"1995","unstructured":"Rose, T., Sebo, P., Bellalou, J. & Ladant, D. Interaction of calcium with Bordetella pertussis adenylate cyclase toxin. Characterization of multiple calcium-binding sites and calcium-induced conformational changes. J Biol Chem 270, 26370\u201326376 (1995).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR27","doi-asserted-by":"crossref","first-page":"16914","DOI":"10.1074\/jbc.M601594200","volume":"281","author":"C Bauche","year":"2006","unstructured":"Bauche, C. et al. Structural and functional characterization of an essential RTX subdomain of Bordetella pertussis adenylate cyclase toxin. J Biol Chem 281, 16914\u201316926 (2006).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR28","doi-asserted-by":"crossref","first-page":"1781","DOI":"10.1074\/jbc.M807312200","volume":"284","author":"A Chenal","year":"2009","unstructured":"Chenal, A., Guijarro, J. I., Raynal, B., Delepierre, M. & Ladant, D. RTX calcium binding motifs are intrinsically disordered in the absence of calcium: implication for protein secretion. J Biol Chem 284, 1781\u20131789 (2009).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR29","doi-asserted-by":"publisher","first-page":"11273","DOI":"10.1021\/bi901447j","volume":"48","author":"GR Szilvay","year":"2009","unstructured":"Szilvay, G. R., Blenner, M. A., Shur, O., Cropek, D. M. & Banta, S. A FRET-based method for probing the conformational behavior of an intrinsically disordered repeat domain from Bordetella pertussis adenylate cyclase. Biochemistry 48, 11273\u201311282, doi: 10.1021\/bi901447j (2009).","journal-title":"Biochemistry"},{"key":"BFsrep42065_CR30","doi-asserted-by":"publisher","first-page":"534","DOI":"10.1016\/j.jmb.2010.01.031","volume":"397","author":"AC Sotomayor Perez","year":"2010","unstructured":"Sotomayor Perez, A. C. et al. Characterization of the regions involved in the calcium-induced folding of the intrinsically disordered RTX motifs from the bordetella pertussis adenylate cyclase toxin. Journal of molecular biology 397, 534\u2013549, doi: 10.1016\/j.jmb.2010.01.031 (2010).","journal-title":"Journal of molecular biology"},{"key":"BFsrep42065_CR31","doi-asserted-by":"publisher","first-page":"3744","DOI":"10.1016\/j.bpj.2010.10.016","volume":"99","author":"A Chenal","year":"2010","unstructured":"Chenal, A. et al. Calcium-induced folding and stabilization of the intrinsically disordered RTX domain of the CyaA toxin. Biophys J 99, 3744\u20133753, doi: 10.1016\/j.bpj.2010.10.016 (2010).","journal-title":"Biophys J"},{"key":"BFsrep42065_CR32","doi-asserted-by":"publisher","first-page":"11929","DOI":"10.1021\/ja404790f","volume":"135","author":"AC Sotomayor-Perez","year":"2013","unstructured":"Sotomayor-Perez, A. C., Subrini, O., Hessel, A., Ladant, D. & Chenal, A. Molecular Crowding Stabilizes Both the Intrinsically Disordered Calcium-Free State and the Folded Calcium-Bound State of a Repeat in Toxin (RTX) Protein. Journal of the American Chemical Society 135, 11929\u201311934, doi: 10.1021\/ja404790f (2013).","journal-title":"Journal of the American Chemical Society"},{"key":"BFsrep42065_CR33","doi-asserted-by":"publisher","first-page":"16997","DOI":"10.1074\/jbc.M110.210393","volume":"286","author":"AC Sotomayor-Perez","year":"2011","unstructured":"Sotomayor-Perez, A. C., Ladant, D. & Chenal, A. Calcium-induced folding of intrinsically disordered repeat-in-toxin (RTX) motifs via changes of protein charges and oligomerization states. J Biol Chem 286, 16997\u201317004, doi: 10.1074\/jbc.M110.210393 (2011).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR34","doi-asserted-by":"publisher","first-page":"14223","DOI":"10.1038\/srep14223","volume":"5","author":"DP O\u2019Brien","year":"2015","unstructured":"O\u2019Brien, D. P. et al. Structural models of intrinsically disordered and calcium-bound folded states of a protein adapted for secretion. Sci Rep 5, 14223, doi: 10.1038\/srep14223 (2015).","journal-title":"Sci Rep"},{"key":"BFsrep42065_CR35","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1080\/09687860500042013","volume":"22","author":"IB Holland","year":"2005","unstructured":"Holland, I. B., Schmitt, L. & Young, J. Type 1 protein secretion in bacteria, the ABC-transporter dependent pathway (review). Mol Membr Biol 22, 29\u201339 (2005).","journal-title":"Mol Membr Biol"},{"key":"BFsrep42065_CR36","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1021\/bi00453a017","volume":"29","author":"HR Masure","year":"1990","unstructured":"Masure, H. R., Au, D. C., Gross, M. K., Donovan, M. G. & Storm, D. R. Secretion of the Bordetella pertussis adenylate cyclase from Escherichia coli containing the hemolysin operon. Biochemistry 29, 140\u2013145 (1990).","journal-title":"Biochemistry"},{"key":"BFsrep42065_CR37","doi-asserted-by":"publisher","first-page":"47","DOI":"10.1016\/j.molcel.2016.03.018","volume":"62","author":"L Bumba","year":"2016","unstructured":"Bumba, L. et al. Calcium-Driven Folding of RTX Domain beta-Rolls Ratchets Translocation of RTX Proteins through Type I Secretion Ducts. Mol Cell 62, 47\u201362, doi: 10.1016\/j.molcel.2016.03.018 (2016).","journal-title":"Mol Cell"},{"key":"BFsrep42065_CR38","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1084\/jem.193.9.1035","volume":"193","author":"P Guermonprez","year":"2001","unstructured":"Guermonprez, P. et al. The adenylate cyclase toxin of Bordetella pertussis binds to target cells via the alpha(M)beta(2) integrin (CD11b\/CD18). J Exp Med 193, 1035\u20131044 (2001).","journal-title":"J Exp Med"},{"key":"BFsrep42065_CR39","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/0014-5793(91)80088-K","volume":"278","author":"IE Ehrmann","year":"1991","unstructured":"Ehrmann, I. E., Gray, M. C., Gordon, V. M., Gray, L. S. & Hewlett, E. L. Hemolytic activity of adenylate cyclase toxin from Bordetella pertussis. FEBS letters 278, 79\u201383, doi: 0014-5793(91)80088-K (1991).","journal-title":"FEBS letters"},{"key":"BFsrep42065_CR40","doi-asserted-by":"crossref","first-page":"22599","DOI":"10.1016\/S0021-9258(18)41715-2","volume":"267","author":"A Rogel","year":"1992","unstructured":"Rogel, A. & Hanski, E. Distinct steps in the penetration of adenylate cyclase toxin of Bordetella pertussis into sheep erythrocytes. Translocation of the toxin across the membrane. J Biol Chem 267, 22599\u201322605 (1992).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR41","doi-asserted-by":"crossref","first-page":"12419","DOI":"10.1074\/jbc.M611226200","volume":"282","author":"M Basler","year":"2007","unstructured":"Basler, M. et al. Segments crucial for membrane translocation and pore-forming activity of Bordetella adenylate cyclase toxin. J Biol Chem 282, 12419\u201312429 (2007).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR42","doi-asserted-by":"publisher","first-page":"10662","DOI":"10.1074\/jbc.M109.089219","volume":"285","author":"JC Eby","year":"2010","unstructured":"Eby, J. C. et al. Selective translocation of the Bordetella pertussis adenylate cyclase toxin across the basolateral membranes of polarized epithelial cells. J Biol Chem 285, 10662\u201310670, doi: 10.1074\/jbc.M109.089219 (2010).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR43","doi-asserted-by":"publisher","first-page":"1317","DOI":"10.1084\/jem.20101558","volume":"208","author":"SR Paccani","year":"2011","unstructured":"Paccani, S. R. et al. The Bordetella pertussis adenylate cyclase toxin binds to T cells via LFA-1 and induces its disengagement from the immune synapse. J Exp Med 208, 1317\u20131330, doi: 10.1084\/jem.20101558 (2011).","journal-title":"J Exp Med"},{"key":"BFsrep42065_CR44","doi-asserted-by":"publisher","first-page":"590","DOI":"10.1111\/mmi.12879","volume":"95","author":"JS Dolores","year":"2015","unstructured":"Dolores, J. S., Agarwal, S., Egerer, M. & Satchell, K. J. Vibrio cholerae MARTX toxin heterologous translocation of beta-lactamase and roles of individual effector domains on cytoskeleton dynamics. Molecular microbiology 95, 590\u2013604, doi: 10.1111\/mmi.12879 (2015).","journal-title":"Molecular microbiology"},{"key":"BFsrep42065_CR45","doi-asserted-by":"crossref","first-page":"14792","DOI":"10.1016\/S0021-9258(18)63769-X","volume":"264","author":"VM Gordon","year":"1989","unstructured":"Gordon, V. M. et al. Adenylate cyclase toxins from Bacillus anthracis and Bordetella pertussis. Different processes for interaction with and entry into target cells. J Biol Chem 264, 14792\u201314796 (1989).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR46","doi-asserted-by":"crossref","first-page":"2755","DOI":"10.1002\/j.1460-2075.1989.tb08417.x","volume":"8","author":"A Rogel","year":"1989","unstructured":"Rogel, A. et al. Bordetella pertussis adenylate cyclase: purification and characterization of the toxic form of the enzyme. Embo J 8, 2755\u20132760 (1989).","journal-title":"Embo J"},{"key":"BFsrep42065_CR47","doi-asserted-by":"crossref","first-page":"1910","DOI":"10.4049\/jimmunol.162.4.1910","volume":"162","author":"P Guermonprez","year":"1999","unstructured":"Guermonprez, P., Ladant, D., Karimova, G., Ullmann, A. & Leclerc, C. Direct delivery of the Bordetella pertussis adenylate cyclase toxin to the MHC class I antigen presentation pathway. J Immunol 162, 1910\u20131916 (1999).","journal-title":"J Immunol"},{"key":"BFsrep42065_CR48","doi-asserted-by":"crossref","first-page":"2831","DOI":"10.1096\/fj.09-131250","volume":"23","author":"J Vojtova-Vodolanova","year":"2009","unstructured":"Vojtova-Vodolanova, J. et al. Oligomerization is involved in pore formation by Bordetella adenylate cyclase toxin. Faseb J 23, 2831\u20132843 (2009).","journal-title":"Faseb J"},{"key":"BFsrep42065_CR49","doi-asserted-by":"publisher","first-page":"e1000901","DOI":"10.1371\/journal.ppat.1000901","volume":"6","author":"L Bumba","year":"2010","unstructured":"Bumba, L., Masin, J., Fiser, R. & Sebo, P. Bordetella adenylate cyclase toxin mobilizes its beta2 integrin receptor into lipid rafts to accomplish translocation across target cell membrane in two steps. PLoS Pathog 6, e1000901, doi: 10.1371\/journal.ppat.1000901 (2010).","journal-title":"PLoS Pathog"},{"key":"BFsrep42065_CR50","doi-asserted-by":"publisher","first-page":"e67648","DOI":"10.1371\/journal.pone.0067648","volume":"8","author":"KB Uribe","year":"2013","unstructured":"Uribe, K. B., Etxebarria, A., Martin, C. & Ostolaza, H. Calpain-Mediated Processing of Adenylate Cyclase Toxin Generates a Cytosolic Soluble Catalytically Active N-Terminal Domain. PLoS One 8, e67648, doi: 10.1371\/journal.pone.0067648 (2013).","journal-title":"PLoS One"},{"key":"BFsrep42065_CR51","doi-asserted-by":"publisher","first-page":"e74248","DOI":"10.1371\/journal.pone.0074248","volume":"8","author":"KB Uribe","year":"2013","unstructured":"Uribe, K. B. et al. Ca2+ influx and tyrosine kinases trigger Bordetella adenylate cyclase toxin (ACT) endocytosis. Cell physiology and expression of the CD11b\/CD18 integrin major determinants of the entry route. PLoS One 8, e74248, doi: 10.1371\/journal.pone.0074248 (2013).","journal-title":"PLoS One"},{"key":"BFsrep42065_CR52","doi-asserted-by":"crossref","first-page":"3760","DOI":"10.1128\/JB.186.12.3760-3765.2004","volume":"186","author":"C Martin","year":"2004","unstructured":"Martin, C. et al. Membrane restructuring by Bordetella pertussis adenylate cyclase toxin, a member of the RTX toxin family. J Bacteriol 186, 3760\u20133765 (2004).","journal-title":"J Bacteriol"},{"key":"BFsrep42065_CR53","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1016\/j.micpath.2008.10.005","volume":"46","author":"GY Cheung","year":"2009","unstructured":"Cheung, G. Y. et al. Functional and structural studies on different forms of the adenylate cyclase toxin of Bordetella pertussis. Microbial pathogenesis 46, 36\u201342 (2009).","journal-title":"Microbial pathogenesis"},{"key":"BFsrep42065_CR54","doi-asserted-by":"crossref","first-page":"1926","DOI":"10.1073\/pnas.73.6.1926","volume":"73","author":"EL Hewlett","year":"1976","unstructured":"Hewlett, E. L., Urban, M. A., Manclark, C. R. & Wolff, J. Extracytoplasmic adenylate cyclase of Bordetella pertussis. Proc Natl Acad Sci USA 73, 1926\u20131930 (1976).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep42065_CR55","doi-asserted-by":"crossref","first-page":"19379","DOI":"10.1016\/S0021-9258(19)47312-2","volume":"264","author":"EL Hewlett","year":"1989","unstructured":"Hewlett, E. L., Gordon, V. M., McCaffery, J. D., Sutherland, W. M. & Gray, M. C. Adenylate cyclase toxin from Bordetella pertussis. Identification and purification of the holotoxin molecule. J Biol Chem 264, 19379\u201319384 (1989).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR56","doi-asserted-by":"crossref","first-page":"12532","DOI":"10.1073\/pnas.95.21.12532","volume":"95","author":"G Karimova","year":"1998","unstructured":"Karimova, G. et al. Charge-dependent translocation of Bordetella pertussis adenylate cyclase toxin into eukaryotic cells: implication for the in vivo delivery of CD8(+) T cell epitopes into antigen-presenting cells. Proc Natl Acad Sci USA 95, 12532\u201312537 (1998).","journal-title":"Proc Natl Acad Sci USA"},{"key":"BFsrep42065_CR57","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1007\/978-1-4614-3704-8_10","volume":"896","author":"JC Karst","year":"2012","unstructured":"Karst, J. C., Sotomayor-Perez, A. C., Ladant, D. & Chenal, A. Estimation of intrinsically disordered protein shape and time-averaged apparent hydration in native conditions by a combination of hydrodynamic methods. Methods in molecular biology 896, 163\u2013177, doi: 10.1007\/978-1-4614-3704-8_10 (2012).","journal-title":"Methods in molecular biology"},{"key":"BFsrep42065_CR58","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1107\/S0021889809043076","volume":"43","author":"H Fischer","year":"2010","unstructured":"Fischer, H., de Oliveira Neto, M., Napolitano, H. B., Polikarpov, I. & Craievich, A. F. Determination of the molecular weight of proteins in solution from a single small-angle X-ray scattering measurement on a relative scale. J. Appl. Cryst. 43, 101\u2013109 (2010).","journal-title":"J. Appl. Cryst."},{"key":"BFsrep42065_CR59","doi-asserted-by":"publisher","first-page":"477","DOI":"10.1038\/nature12070","volume":"496","author":"RP Rambo","year":"2013","unstructured":"Rambo, R. P. & Tainer, J. A. Accurate assessment of mass, models and resolution by small-angle scattering. Nature 496, 477\u2013+, doi: 10.1038\/nature12070 (2013).","journal-title":"Nature"},{"key":"BFsrep42065_CR60","doi-asserted-by":"publisher","first-page":"2879","DOI":"10.1016\/S0006-3495(99)77443-6","volume":"76","author":"DI Svergun","year":"1999","unstructured":"Svergun, D. I. Restoring low resolution structure of biological macromolecules from solution scattering using simulated annealing. Biophysical Journal 76, 2879\u20132886, doi: 10.1016\/S0006-3495(99)77443-6 (1999).","journal-title":"Biophysical Journal"},{"key":"BFsrep42065_CR61","doi-asserted-by":"publisher","first-page":"860","DOI":"10.1107\/S0021889803000268","volume":"36","author":"VV Volkov","year":"2003","unstructured":"Volkov, V. V. & Svergun, D. I. Uniqueness of ab initio shape determination in small-angle scattering. J Appl Crystallogr 36, 860\u2013864, doi: 10.1107\/S0021889803000268 (2003).","journal-title":"J Appl Crystallogr"},{"key":"BFsrep42065_CR62","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1111\/j.1365-2958.1991.tb00723.x","volume":"5","author":"RA Welch","year":"1991","unstructured":"Welch, R. A. Pore-forming cytolysins of gram-negative bacteria. Molecular microbiology 5, 521\u2013528 (1991).","journal-title":"Molecular microbiology"},{"key":"BFsrep42065_CR63","unstructured":"Chenal, A., Sotomayor Perez, A. C. & Ladant, D. In The Comprehensive Sourcebook of Bacterial Protein Toxins, 4th Edition (Elsevier, 2015)."},{"key":"BFsrep42065_CR64","doi-asserted-by":"publisher","first-page":"1","DOI":"10.3390\/toxins7010001","volume":"7","author":"AC Sotomayor-Perez","year":"2015","unstructured":"Sotomayor-Perez, A. C., Ladant, D. & Chenal, A. Disorder-to-order transition in the CyaA toxin RTX domain: implications for toxin secretion. Toxins (Basel) 7, 1\u201320, doi: 10.3390\/toxins7010001 (2015).","journal-title":"Toxins (Basel)"},{"key":"BFsrep42065_CR65","doi-asserted-by":"crossref","first-page":"800","DOI":"10.1046\/j.1432-1327.2001.01937.x","volume":"268","author":"C Schindel","year":"2001","unstructured":"Schindel, C. et al. Interaction of Escherichia coli hemolysin with biological membranes. A study using cysteine scanning mutagenesis. European journal of biochemistry\/FEBS 268, 800\u2013808 (2001).","journal-title":"European journal of biochemistry\/FEBS"},{"key":"BFsrep42065_CR66","doi-asserted-by":"publisher","first-page":"1225","DOI":"10.1016\/j.bbamem.2010.03.007","volume":"1798","author":"L Sanchez-Magraner","year":"2010","unstructured":"Sanchez-Magraner, L. et al. Interdomain Ca(2+) effects in Escherichia coli alpha-haemolysin: Ca(2+) binding to the C-terminal domain stabilizes both C- and N-terminal domains. Biochim Biophys Acta 1798, 1225\u20131233, doi: 10.1016\/j.bbamem.2010.03.007 (2010).","journal-title":"Biochim Biophys Acta"},{"key":"BFsrep42065_CR67","doi-asserted-by":"publisher","first-page":"ftv075","DOI":"10.1093\/femspd\/ftv075","volume":"73","author":"J Masin","year":"2015","unstructured":"Masin, J., Osicka, R., Bumba, L. & Sebo, P. Bordetella adenylate cyclase toxin: a unique combination of a pore-forming moiety with a cell-invading adenylate cyclase enzyme. Pathog Dis 73, ftv075, doi: 10.1093\/femspd\/ftv075 (2015).","journal-title":"Pathog Dis"},{"key":"BFsrep42065_CR68","doi-asserted-by":"crossref","first-page":"9695","DOI":"10.1074\/jbc.270.17.9695","volume":"270","author":"AS Otero","year":"1995","unstructured":"Otero, A. S., Yi, X. B., Gray, M. C., Szabo, G. & Hewlett, E. L. Membrane depolarization prevents cell invasion by Bordetella pertussis adenylate cyclase toxin. J Biol Chem 270, 9695\u20139697 (1995).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR69","doi-asserted-by":"crossref","first-page":"2808","DOI":"10.1074\/jbc.M609979200","volume":"282","author":"R Fiser","year":"2007","unstructured":"Fiser, R. et al. Third activity of Bordetella adenylate cyclase (AC) toxin-hemolysin. Membrane translocation of AC domain polypeptide promotes calcium influx into CD11b+ monocytes independently of the catalytic and hemolytic activities. J Biol Chem 282, 2808\u20132820 (2007).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR70","doi-asserted-by":"publisher","first-page":"e1002580","DOI":"10.1371\/journal.ppat.1002580","volume":"8","author":"R Fiser","year":"2012","unstructured":"Fiser, R. et al. Calcium influx rescues adenylate cyclase-hemolysin from rapid cell membrane removal and enables phagocyte permeabilization by toxin pores. PLoS Pathog 8, e1002580, doi: 10.1371\/journal.ppat.1002580 (2012).","journal-title":"PLoS Pathog"},{"key":"BFsrep42065_CR71","doi-asserted-by":"crossref","first-page":"3154","DOI":"10.1016\/S0021-9258(18)49967-X","volume":"266","author":"A Rogel","year":"1991","unstructured":"Rogel, A., Meller, R. & Hanski, E. Adenylate cyclase toxin from Bordetella pertussis. The relationship between induction of cAMP and hemolysis. J Biol Chem 266, 3154\u20133161 (1991).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR72","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1128\/iai.58.6.1959-1964.1990","volume":"58","author":"DF Boehm","year":"1990","unstructured":"Boehm, D. F., Welch, R. A. & Snyder, I. S. Domains of Escherichia coli hemolysin (HlyA) involved in binding of calcium and erythrocyte membranes. Infection and immunity 58, 1959\u20131964 (1990).","journal-title":"Infection and immunity"},{"key":"BFsrep42065_CR73","doi-asserted-by":"crossref","first-page":"454","DOI":"10.1111\/j.1432-1033.1996.0454h.x","volume":"240","author":"A Dobereiner","year":"1996","unstructured":"Dobereiner, A., Schmid, A., Ludwig, A., Goebel, W. & Benz, R. The effects of calcium and other polyvalent cations on channel formation by Escherichia coli alpha-hemolysin in red blood cells and lipid bilayer membranes. European journal of biochemistry\/FEBS 240, 454\u2013460 (1996).","journal-title":"European journal of biochemistry\/FEBS"},{"key":"BFsrep42065_CR74","first-page":"876","volume":"12","author":"TJ Schoenmakers","year":"1992","unstructured":"Schoenmakers, T. J., Visser, G. J., Flik, G. & Theuvenet, A. P. CHELATOR: an improved method for computing metal ion concentrations in physiological solutions. Biotechniques 12, 870\u2013874, 876\u2013879 (1992).","journal-title":"Biotechniques"},{"key":"BFsrep42065_CR75","doi-asserted-by":"publisher","first-page":"331","DOI":"10.1007\/978-1-4614-3704-8_22","volume":"896","author":"AC Sotomayor-Perez","year":"2012","unstructured":"Sotomayor-Perez, A. C., Karst, J. C., Ladant, D. & Chenal, A. Mean net charge of intrinsically disordered proteins: experimental determination of protein valence by electrophoretic mobility measurements. Methods in molecular biology 896, 331\u2013349, doi: 10.1007\/978-1-4614-3704-8_22 (2012).","journal-title":"Methods in molecular biology"},{"key":"BFsrep42065_CR76","doi-asserted-by":"publisher","first-page":"24239","DOI":"10.1074\/jbc.M703392200","volume":"282","author":"A Perier","year":"2007","unstructured":"Perier, A. et al. Concerted protonation of key histidines triggers membrane interaction of the diphtheria toxin T domain. J Biol Chem 282, 24239\u201324245, doi: 10.1074\/jbc.M703392200 (2007).","journal-title":"J Biol Chem"},{"key":"BFsrep42065_CR77","doi-asserted-by":"crossref","first-page":"890","DOI":"10.1016\/j.jmb.2005.04.036","volume":"349","author":"A Chenal","year":"2005","unstructured":"Chenal, A. et al. Conformational states and thermodynamics of alpha-lactalbumin bound to membranes: a case study of the effects of pH, calcium, lipid membrane curvature and charge. Journal of molecular biology 349, 890\u2013905 (2005).","journal-title":"Journal of molecular biology"},{"key":"BFsrep42065_CR78","doi-asserted-by":"crossref","first-page":"1277","DOI":"10.1107\/S0021889803012779","volume":"36","author":"PV Konarev","year":"2003","unstructured":"Konarev, P. V., Volkov, V. V., Sokolova, A. V., Koch, M. H. J. & Svergun, D. I. PRIMUS - a Windows-PC based system for small-angle scattering data analysis. J. Appl. Crystallogr. 36, 1277\u20131282 (2003).","journal-title":"J. Appl. Crystallogr."},{"key":"BFsrep42065_CR79","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1107\/S0021889800014126","volume":"34","author":"MB Kozin","year":"2001","unstructured":"Kozin, M. B. & Svergun, D. I. Automated matching of high- and low-resolution structural models. J. Appl. Cryst. 34, 33\u201341 (2001).","journal-title":"J. Appl. Cryst."},{"key":"BFsrep42065_CR80","doi-asserted-by":"publisher","first-page":"11992","DOI":"10.1074\/jbc.M809342200","volume":"284","author":"N Leulliot","year":"2009","unstructured":"Leulliot, N. et al. The family X DNA polymerase from Deinococcus radiodurans adopts a non-standard extended conformation. J Biol Chem 284, 11992\u201311999, doi: 10.1074\/jbc.M809342200 (2009).","journal-title":"J Biol Chem"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/srep42065","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep42065.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/srep42065.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,22]],"date-time":"2024-06-22T08:00:46Z","timestamp":1719043246000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/srep42065"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,2,10]]},"references-count":80,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,2,27]]}},"alternative-id":["BFsrep42065"],"URL":"https:\/\/doi.org\/10.1038\/srep42065","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,2,10]]},"assertion":[{"value":"12 August 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 January 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 February 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing financial interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"42065"}}