{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T14:06:25Z","timestamp":1790431585608,"version":"4.1.0"},"reference-count":44,"publisher":"MDPI AG","issue":"6","license":[{"start":{"date-parts":[[2018,6,16]],"date-time":"2018-06-16T00:00:00Z","timestamp":1529107200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["16-05919S (J.M.), 18-18079S (R.O.) and 18-20621S (P.S.)"],"award-info":[{"award-number":["16-05919S (J.M.), 18-18079S (R.O.) and 18-20621S (P.S.)"]}],"id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Ministry of Education, Youth and Sports of the Czech Republic","award":["LM2015064"],"award-info":[{"award-number":["LM2015064"]}]},{"DOI":"10.13039\/501100003243","name":"Ministerstvo Zdravotnictv\u00ed Cesk\u00e9 Republiky","doi-asserted-by":"publisher","award":["NV16-28126A"],"award-info":[{"award-number":["NV16-28126A"]}],"id":[{"id":"10.13039\/501100003243","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>The adenylate cyclase toxin-hemolysin (CyaA, ACT, or AC-Hly) plays a crucial role in virulence and airway colonization capacity of the whooping cough agent Bordetella pertussis. The toxin penetrates target cell membranes and exhibits three distinct biological activities. A population of CyaA conformers forms small cation-selective pores that permeabilize the cell membrane for potassium efflux, which can provoke colloid-osmotic (oncotic) cell lysis. The other two activities are due to CyaA conformers that transiently form calcium influx conduits in the target cell membrane and translocate the adenylate cyclase (AC) enzyme into cytosol of cells. A fourth putative biological activity has recently been reported; an intrinsic phospholipase A (PLA) activity was claimed to be associated with the CyaA polypeptide and be involved in the mechanism of translocation of the AC enzyme polypeptide across cell membrane lipid bilayer. However, the conclusions drawn by the authors contradicted their own results and we show them to be erroneous. We demonstrate that highly purified CyaA is devoid of any detectable phospholipase A1 activity and that contrary to the published claims, the two putative conserved phospholipase A catalytic residues, namely the Ser606 and Asp1079 residues, are not involved in the process of membrane translocation of the AC domain of CyaA across target membranes.<\/jats:p>","DOI":"10.3390\/toxins10060245","type":"journal-article","created":{"date-parts":[[2018,6,18]],"date-time":"2018-06-18T10:57:11Z","timestamp":1529319431000},"page":"245","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":7,"title":["Bordetella Pertussis Adenylate Cyclase Toxin Does Not Possess a Phospholipase A Activity; Serine 606 and Aspartate 1079 Residues Are Not Involved in Target Cell Delivery of the Adenylyl Cyclase Enzyme Domain"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6659-5447","authenticated-orcid":false,"given":"Ladislav","family":"Bumba","sequence":"first","affiliation":[{"name":"Institute of Microbiology of the CAS, v.v.i., 142 20 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jiri","family":"Masin","sequence":"additional","affiliation":[{"name":"Institute of Microbiology of the CAS, v.v.i., 142 20 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Adriana","family":"Osickova","sequence":"additional","affiliation":[{"name":"Institute of Microbiology of the CAS, v.v.i., 142 20 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Radim","family":"Osicka","sequence":"additional","affiliation":[{"name":"Institute of Microbiology of the CAS, v.v.i., 142 20 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peter","family":"Sebo","sequence":"additional","affiliation":[{"name":"Institute of Microbiology of the CAS, v.v.i., 142 20 Prague, Czech Republic"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,6,16]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"326","DOI":"10.1128\/CMR.18.2.326-382.2005","article-title":"Molecular pathogenesis, epidemiology, and clinical manifestations of respiratory infections due to Bordetella pertussis and other Bordetella subspecies","volume":"18","author":"Mattoo","year":"2005","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.molcel.2016.03.018","article-title":"Calcium-driven folding of RTX domain beta-rolls ratchets translocation of RTX proteins through type I secretion ducts","volume":"62","author":"Bumba","year":"2016","journal-title":"Mol. Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1076","DOI":"10.1111\/j.1574-6976.2010.00231.x","article-title":"RTX proteins: A highly diverse family secreted by a common mechanism","volume":"34","author":"Linhartova","year":"2010","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","unstructured":"Novak, J., Cerny, O., Osickova, A., Linhartova, I., Masin, J., Bumba, L., Sebo, P., and Osicka, R. (2017). Structure-function relationships underlying the capacity of Bordetella adenylate cyclase toxin to disarm host phagocytes. Toxins, 9.","DOI":"10.3390\/toxins9100300"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"29137","DOI":"10.1038\/srep29137","article-title":"Negatively charged residues of the segment linking the enzyme and cytolysin moieties restrict the membrane-permeabilizing capacity of adenylate cyclase toxin","volume":"6","author":"Masin","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"32585","DOI":"10.1074\/jbc.M113.508838","article-title":"Characterization of a membrane-active peptide from the Bordetella pertussis CyaA toxin","volume":"288","author":"Subrini","year":"2013","journal-title":"J. Biol. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","unstructured":"Voegele, A., Subrini, O., Sapay, N., Ladant, D., and Chenal, A. (2017). Membrane-active properties of an amphitropic peptide from the CyaA toxin translocation region. Toxins, 9.","DOI":"10.3390\/toxins9110369"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Masin, J., Osicka, R., Bumba, L., and Sebo, P. (2015). Bordetella adenylate cyclase toxin: A unique combination of a pore-forming moiety with a cell-invading adenylate cyclase enzyme. Pathog. Dis., 73.","DOI":"10.1093\/femspd\/ftv075"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"433","DOI":"10.1126\/science.7939682","article-title":"Internal lysine palmitoylation in adenylate cyclase toxin from Bordetella pertussis","volume":"266","author":"Hackett","year":"1994","journal-title":"Science"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1035","DOI":"10.1084\/jem.193.9.1035","article-title":"The adenylate cyclase toxin of Bordetella pertussis binds to target cells via the alpha(M)\u03b2(2) integrin (CD11b\/CD18)","volume":"193","author":"Guermonprez","year":"2001","journal-title":"J. Exp. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"e10766","DOI":"10.7554\/eLife.10766","article-title":"Bordetella adenylate cyclase toxin is a unique ligand of the integrin complement receptor 3","volume":"4","author":"Osicka","year":"2015","journal-title":"Elife"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.febslet.2014.12.023","article-title":"Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding","volume":"589","author":"Hasan","year":"2015","journal-title":"FEBS Lett."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5355","DOI":"10.1073\/pnas.0711400105","article-title":"RTX cytotoxins recognize \u03b22 integrin receptors through n-linked oligosaccharides","volume":"105","author":"Morova","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1066","DOI":"10.1128\/iai.56.5.1066-1069.1988","article-title":"Inhibitors of receptor-mediated endocytosis block the entry of Bacillus anthracis adenylate cyclase toxin but not that of Bordetella pertussis adenylate cyclase toxin","volume":"56","author":"Gordon","year":"1988","journal-title":"Infect. Immun."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"17503","DOI":"10.1016\/S0021-9258(19)47400-0","article-title":"Adenylate cyclase toxin from Bordetella pertussis. Conformational change associated with toxin activity","volume":"266","author":"Hewlett","year":"1991","journal-title":"J. Biol. Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1181","DOI":"10.1128\/IAI.05711-11","article-title":"Delivery of large heterologous polypeptides across the cytoplasmic membrane of antigen-presenting cells by the Bordetella RTX hemolysin moiety lacking the adenylyl cyclase domain","volume":"80","author":"Holubova","year":"2012","journal-title":"Infect. Immun."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"8077","DOI":"10.1021\/bi034295f","article-title":"Channel formation in model membranes by the adenylate cyclase toxin of Bordetella pertussis: Effect of calcium","volume":"42","author":"Knapp","year":"2003","journal-title":"Biochemistry"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"26370","DOI":"10.1074\/jbc.270.44.26370","article-title":"Interaction of calcium with Bordetella pertussis adenylate cyclase toxin. Characterization of multiple calcium-binding sites and calcium-induced conformational changes","volume":"270","author":"Rose","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Wald, T., Osickova, A., Masin, J., Liskova, P.M., Petry-Podgorska, I., Matousek, T., Sebo, P., and Osicka, R. (2016). Transmembrane segments of complement receptor 3 do not participate in cytotoxic activities but determine receptor structure required for action of Bordetella adenylate cyclase toxin. Pathog. Dis., 74.","DOI":"10.1093\/femspd\/ftw008"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"42065","DOI":"10.1038\/srep42065","article-title":"Stability, structural and functional properties of a monomeric, calcium-loaded adenylate cyclase toxin, CyaA, from Bordetella pertussis","volume":"7","author":"Cannella","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3242","DOI":"10.1128\/iai.58.10.3242-3247.1990","article-title":"Deletions affecting hemolytic and toxin activities of Bordetella pertussis adenylate cyclase","volume":"58","author":"Bellalou","year":"1990","journal-title":"Infect. Immun."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3841","DOI":"10.1073\/pnas.77.7.3841","article-title":"Calmodulin activates prokaryotic adenylate cyclase","volume":"77","author":"Wolff","year":"1980","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Fiser, R., Masin, J., Bumba, L., Pospisilova, E., Fayolle, C., Basler, M., Sadilkova, L., Adkins, I., Kamanova, J., and Cerny, J. (2012). Calcium influx rescues adenylate cyclase-hemolysin from rapid cell membrane removal and enables phagocyte permeabilization by toxin pores. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002580"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"18260","DOI":"10.1074\/jbc.273.29.18260","article-title":"Distinct mechanisms for K+ efflux, intoxication, and hemolysis by Bordetella pertussis AC toxin","volume":"273","author":"Gray","year":"1998","journal-title":"J. Biol. Chem."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2831","DOI":"10.1096\/fj.09-131250","article-title":"Oligomerization is involved in pore formation by Bordetella adenylate cyclase toxin","volume":"23","author":"Basler","year":"2009","journal-title":"FASEB J."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1016\/j.ab.2013.10.039","article-title":"Quantification of potassium levels in cells treated with Bordetella adenylate cyclase toxin","volume":"450","author":"Wald","year":"2014","journal-title":"Anal. Biochem."},{"key":"ref_27","first-page":"2808","article-title":"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","volume":"282","author":"Fiser","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Bumba, L., Masin, J., Fiser, R., and Sebo, P. (2010). Bordetella adenylate cyclase toxin mobilizes its \u03b22 integrin receptor into lipid rafts to accomplish translocation across target cell membrane in two steps. PLoS Pathog., 6.","DOI":"10.1371\/journal.ppat.1000901"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"9695","DOI":"10.1074\/jbc.270.17.9695","article-title":"Membrane depolarization prevents cell invasion by Bordetella pertussis adenylate cyclase toxin","volume":"270","author":"Otero","year":"1995","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"20473","DOI":"10.1073\/pnas.1312975110","article-title":"Bordetella pertussis adenylate cyclase toxin translocation across a tethered lipid bilayer","volume":"110","author":"Veneziano","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"5752","DOI":"10.1073\/pnas.95.10.5752","article-title":"A bacterial two-hybrid system based on a reconstituted signal transduction pathway","volume":"95","author":"Karimova","year":"1998","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"12419","DOI":"10.1074\/jbc.M611226200","article-title":"Segments crucial for membrane translocation and pore-forming activity of Bordetella adenylate cyclase toxin","volume":"282","author":"Basler","year":"2007","journal-title":"J. Biol. Chem."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"2207","DOI":"10.1128\/IAI.74.4.2207-2214.2006","article-title":"Pore-forming and enzymatic activities of Bordetella pertussis adenylate cyclase toxin synergize in promoting lysis of monocytes","volume":"74","author":"Basler","year":"2006","journal-title":"Infect. Immun."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"9330","DOI":"10.1038\/s41598-017-09575-6","article-title":"The conserved tyrosine residue 940 plays a key structural role in membrane interaction of Bordetella adenylate cyclase toxin","volume":"7","author":"Masin","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"37644","DOI":"10.1016\/S0021-9258(19)52940-4","article-title":"An amphipathic alpha-helix including glutamates 509 and 516 is crucial for membrane translocation of adenylate cyclase toxin and modulates formation and cation selectivity of its membrane channels","volume":"274","author":"Osickova","year":"1999","journal-title":"J. Biol. Chem."},{"key":"ref_36","first-page":"E6784","article-title":"Phospholipase a activity of adenylate cyclase toxin mediates translocation of its adenylate cyclase domain","volume":"114","author":"Uribe","year":"2017","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Masin, J., Osicka, R., Bumba, L., and Sebo, P. Phospholipase a activity of adenylate cyclase toxin?. Proc. Natl. Acad. Sci. USA, 2018.","DOI":"10.1073\/pnas.1722588115"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1128\/IAI.68.1.247-256.2000","article-title":"Delivery of CD8(+) T-cell epitopes into major histocompatibility complex class I antigen presentation pathway by Bordetella pertussis adenylate cyclase: Delineation of cell invasive structures and permissive insertion sites","volume":"68","author":"Osicka","year":"2000","journal-title":"Infect. Immun."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"447","DOI":"10.1111\/j.1365-2958.2005.04958.x","article-title":"Macrophage cytotoxicity produced by adenylate cyclase toxin from Bordetella pertussis: More than just making cyclic AMP!","volume":"59","author":"Hewlett","year":"2006","journal-title":"Mol. Microbiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1111\/j.1365-2958.1995.mmi_17061015.x","article-title":"Identification by in vitro complementation of regions required for cell-invasive activity of Bordetella pertussis adenylate cyclase toxin","volume":"17","author":"Iwaki","year":"1995","journal-title":"Mol. Microbiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"38514","DOI":"10.1074\/jbc.M304387200","article-title":"Interaction of Bordetella pertussis adenylate cyclase with CD11b\/CD18: Role of toxin acylation and identification of the main integrin interaction domain","volume":"278","author":"Bauche","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"144","DOI":"10.1016\/j.bbamem.2003.11.008","article-title":"Different structural requirements for adenylate cyclase toxin interactions with erythrocyte and liposome membranes","volume":"1660","author":"Masin","year":"2004","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1128\/iai.55.1.135-140.1987","article-title":"Bordetella pertussis adenylate cyclase: Effects of affinity-purified adenylate cyclase on human polymorphonuclear leukocyte functions","volume":"55","author":"Friedman","year":"1987","journal-title":"Infect. Immun."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2612","DOI":"10.1016\/S0021-9258(18)69110-0","article-title":"Interaction of Bordetella pertussis adenylate cyclase with calmodulin. Identification of two separated calmodulin-binding domains","volume":"263","author":"Ladant","year":"1988","journal-title":"J. Biol. Chem."}],"container-title":["Toxins"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6651\/10\/6\/245\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:09:00Z","timestamp":1760195340000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6651\/10\/6\/245"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,6,16]]},"references-count":44,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2018,6]]}},"alternative-id":["toxins10060245"],"URL":"https:\/\/doi.org\/10.3390\/toxins10060245","relation":{},"ISSN":["2072-6651"],"issn-type":[{"value":"2072-6651","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,6,16]]}}}