This page has only limited features, please log in for full access.

Prof. Peter SEBO
Institute of Microbiology of the Czech Academy of Sciences

Basic Info


Research Keywords & Expertise

0 Innate Immunity
0 Vaccines
0 bordetella pertussis
0 bacterial toxins
0 molecular and cellular microbiology

Fingerprints

bordetella pertussis
Vaccines
Innate Immunity
bacterial toxins

Honors and Awards

The user has no records in this section


Career Timeline

The user has no records in this section.


Short Biography

The user biography is not available.
Following
Followers
Co Authors
The list of users this user is following is empty.
Following: 0 users

Feed

Journal article
Published: 23 August 2021 in International Journal of Molecular Sciences
Reads 0
Downloads 0

The mucus layer protects airway epithelia from damage by noxious agents. Intriguingly, Bordetella pertussis bacteria provoke massive mucus production by nasopharyngeal epithelia during the initial coryza-like catarrhal stage of human pertussis and the pathogen transmits in mucus-containing aerosol droplets expelled by sneezing and post-nasal drip-triggered cough. We investigated the role of the cAMP-elevating adenylate cyclase (CyaA) and pertussis (PT) toxins in the upregulation of mucin production in B. pertussis-infected airway epithelia. Using human pseudostratified airway epithelial cell layers cultured at air–liquid interface (ALI), we show that purified CyaA and PT toxins (100 ng/mL) can trigger production of the major airway mucins Muc5AC and Muc5B. Upregulation of mucin secretion involved activation of the cAMP response element binding protein (CREB) and was blocked by the 666-15-Calbiochem inhibitor of CREB-mediated gene transcription. Intriguingly, a B. pertussis mutant strain secreting only active PT and producing the enzymatically inactive CyaA-AC toxoid failed to trigger any important mucus production in infected epithelial cell layers in vitro or in vivo in the tracheal epithelia of intranasally infected mice. In contrast, the PT toxoid-producing B. pertussis mutant secreting the active CyaA toxin elicited a comparable mucin production as infection of epithelial cell layers or tracheal epithelia of infected mice by the wild-type B. pertussis secreting both PT and CyaA toxins. Hence, the cAMP-elevating activity of B. pertussis-secreted CyaA was alone sufficient for activation of mucin production through a CREB-dependent mechanism in B. pertussis-infected airway epithelia in vivo.

ACS Style

Anna Malandra; Waheed Ur Rahman; Nela Klimova; Gaia Streparola; Jana Holubova; Adriana Osickova; Simone Bariselli; Peter Sebo; Radim Osicka. Bordetella Adenylate Cyclase Toxin Elicits Airway Mucin Secretion through Activation of the cAMP Response Element Binding Protein. International Journal of Molecular Sciences 2021, 22, 9064 .

AMA Style

Anna Malandra, Waheed Ur Rahman, Nela Klimova, Gaia Streparola, Jana Holubova, Adriana Osickova, Simone Bariselli, Peter Sebo, Radim Osicka. Bordetella Adenylate Cyclase Toxin Elicits Airway Mucin Secretion through Activation of the cAMP Response Element Binding Protein. International Journal of Molecular Sciences. 2021; 22 (16):9064.

Chicago/Turabian Style

Anna Malandra; Waheed Ur Rahman; Nela Klimova; Gaia Streparola; Jana Holubova; Adriana Osickova; Simone Bariselli; Peter Sebo; Radim Osicka. 2021. "Bordetella Adenylate Cyclase Toxin Elicits Airway Mucin Secretion through Activation of the cAMP Response Element Binding Protein." International Journal of Molecular Sciences 22, no. 16: 9064.

Journal article
Published: 26 May 2021 in Journal of Biological Chemistry
Reads 0
Downloads 0

The whooping cough agent Bordetella pertussis secretes an adenylate cyclase toxin (CyaA) that through its large carboxy-proximal Repeat-in-ToXin (RTX) domain binds the complement receptor 3 (CR3). The RTX domain consists of five blocks (I–V) of characteristic glycine and aspartate-rich nonapeptides that fold into five Ca2+-loaded parallel β-rolls. Previous work indicated that the CR3-binding structure comprises the interface of β-rolls II and III. To test if further portions of the RTX domain contribute to CR3 binding, we generated a construct with the RTX block II/III interface (CyaA residues 1132–1294) linked directly to the C-terminal block V fragment bearing the folding scaffold (CyaA residues 1562–1681). Despite deletion of 267 internal residues of the RTX domain, the Ca2+-driven folding of the hybrid block III/V β-roll still supported formation of the CR3-binding structure at the interface of β-rolls II and III. Moreover, upon stabilization by N- and C-terminal flanking segments, the block III/V hybrid-comprising constructs competed with CyaA for CR3 binding and induced formation of CyaA toxin-neutralizing antibodies in mice. Finally, a truncated CyaAΔ1295-1561 toxin bound and penetrated erythrocytes and CR3-expressing cells, showing that the deleted portions of RTX blocks III, IV, and V (residues 1295–1561) were dispensable for CR3 binding and for toxin translocation across the target cell membrane. This suggests that almost a half of the RTX domain of CyaA is not involved in target cell interaction and rather serves the purpose of toxin secretion.

ACS Style

Carlos Angel Espinosa-Vinals; Jiri Masin; Jana Holubova; Ondrej Stanek; David Jurnecka; Radim Osicka; Peter Sebo; Ladislav Bumba. Almost half of the RTX domain is dispensable for complement receptor 3 binding and cell-invasive activity of the Bordetella adenylate cyclase toxin. Journal of Biological Chemistry 2021, 297, 100833 .

AMA Style

Carlos Angel Espinosa-Vinals, Jiri Masin, Jana Holubova, Ondrej Stanek, David Jurnecka, Radim Osicka, Peter Sebo, Ladislav Bumba. Almost half of the RTX domain is dispensable for complement receptor 3 binding and cell-invasive activity of the Bordetella adenylate cyclase toxin. Journal of Biological Chemistry. 2021; 297 (1):100833.

Chicago/Turabian Style

Carlos Angel Espinosa-Vinals; Jiri Masin; Jana Holubova; Ondrej Stanek; David Jurnecka; Radim Osicka; Peter Sebo; Ladislav Bumba. 2021. "Almost half of the RTX domain is dispensable for complement receptor 3 binding and cell-invasive activity of the Bordetella adenylate cyclase toxin." Journal of Biological Chemistry 297, no. 1: 100833.

Journal article
Published: 27 January 2021 in Diagnostics
Reads 0
Downloads 0

Serological diagnosis of pertussis is mainly based on anti-pertussis toxin (PT) IgG antibodies. Since PT is included in all acellular vaccines (ACV), serological assays do not differentiate antibodies induced by ACVs and infection. Adenylate cyclase toxin (ACT) is not included in the ACVs, which makes it a promising candidate for pertussis serology with the specific aim of separating infection- and ACV-induced antibodies. A multiplex lateral flow test with PT and ACT antigens was developed to measure serum antibodies from pertussis-seropositive patients (n = 46), healthy controls (n = 102), and subjects who received a booster dose of ACV containing PT, filamentous hemagglutinin, and pertactin (n = 67) with paired sera collected before and one month after the vaccination. If the diagnosis was solely based on anti-PT antibodies, 98.5–44.8% specificity (before and after vaccination, respectively) and 78.2% sensitivity were achieved, whereas if ACT was used in combination with PT, the sensitivity of the assay increased to 91.3% without compromising specificity. No increase in the level of anti-ACT antibodies was found after vaccination. This exploratory study indicates that the use of ACT for serology would be beneficial in combination with a lower quantitative cutoff for anti-PT antibodies, and particularly in children and adolescents who frequently receive booster vaccinations.

ACS Style

Aapo Knuutila; Alex-Mikael Barkoff; Jussi Mertsola; Radim Osicka; Peter Sebo; Qiushui He. Simultaneous Determination of Antibodies to Pertussis Toxin and Adenylate Cyclase Toxin Improves Serological Diagnosis of Pertussis. Diagnostics 2021, 11, 180 .

AMA Style

Aapo Knuutila, Alex-Mikael Barkoff, Jussi Mertsola, Radim Osicka, Peter Sebo, Qiushui He. Simultaneous Determination of Antibodies to Pertussis Toxin and Adenylate Cyclase Toxin Improves Serological Diagnosis of Pertussis. Diagnostics. 2021; 11 (2):180.

Chicago/Turabian Style

Aapo Knuutila; Alex-Mikael Barkoff; Jussi Mertsola; Radim Osicka; Peter Sebo; Qiushui He. 2021. "Simultaneous Determination of Antibodies to Pertussis Toxin and Adenylate Cyclase Toxin Improves Serological Diagnosis of Pertussis." Diagnostics 11, no. 2: 180.

Journal article
Published: 22 December 2020 in mSystems
Reads 0
Downloads 0

We show that a spontaneous mutation that upregulates transcription of an operon encoding ribosomal proteins and causes overproduction of the downstream-encoded α subunit (RpoA) of RNA polymerase causes global effects on gene expression levels and proteome composition of Bordetella pertussis . Nevertheless, the resulting important downregulation of the BvgAS-controlled expression of virulence factors of the whooping cough agent did not compromise its capacity to persist for prolonged periods inside primary human macrophage cells, and it even enhanced its capacity to persist in infected mouse lungs.

ACS Style

Jakub Novák; David Jurnečka; Irena Linhartová; Jana Holubová; Ondřej Staněk; Daniel Štipl; Ana Dienstbier; Branislav Večerek; Nayara Azevedo; Jan Provazník; Vladimír Beneš; Peter Šebo. A Mutation Upstream of the rplN-rpsD Ribosomal Operon Downregulates Bordetella pertussis Virulence Factor Production without Compromising Bacterial Survival within Human Macrophages. mSystems 2020, 5, 1 .

AMA Style

Jakub Novák, David Jurnečka, Irena Linhartová, Jana Holubová, Ondřej Staněk, Daniel Štipl, Ana Dienstbier, Branislav Večerek, Nayara Azevedo, Jan Provazník, Vladimír Beneš, Peter Šebo. A Mutation Upstream of the rplN-rpsD Ribosomal Operon Downregulates Bordetella pertussis Virulence Factor Production without Compromising Bacterial Survival within Human Macrophages. mSystems. 2020; 5 (6):1.

Chicago/Turabian Style

Jakub Novák; David Jurnečka; Irena Linhartová; Jana Holubová; Ondřej Staněk; Daniel Štipl; Ana Dienstbier; Branislav Večerek; Nayara Azevedo; Jan Provazník; Vladimír Beneš; Peter Šebo. 2020. "A Mutation Upstream of the rplN-rpsD Ribosomal Operon Downregulates Bordetella pertussis Virulence Factor Production without Compromising Bacterial Survival within Human Macrophages." mSystems 5, no. 6: 1.

Journal article
Published: 19 November 2020 in Vaccines
Reads 0
Downloads 0

Bordetella pertussis whole-cell vaccines (wP) caused a spectacular drop of global pertussis incidence, but since the replacement of wP with acellular pertussis vaccines (aP), pertussis has resurged in developed countries within 7 to 12 years of the change from wP to aP. In the mouse infection model, we examined whether addition of further protective antigens into the aP vaccine, such as type 2 and type 3 fimbriae (FIM2/3) with outer membrane lipooligosaccharide (LOS) and/or of the adenylate cyclase toxoid (dACT), which elicits antibodies neutralizing the CyaA toxin, could enhance the capacity of the aP vaccine to prevent colonization of the nasal mucosa by B. pertussis. The addition of the toxoid and of the opsonizing antibody-inducing agglutinogens modestly enhanced the already high capacity of intraperitoneally-administered aP vaccine to elicit sterilizing immunity, protecting mouse lungs from B. pertussis infection. At the same time, irrespective of FIM2/3 with LOS and dACT addition, the aP vaccination ablated the natural capacity of BALB/c mice to clear B. pertussis infection from the nasal cavity. While wP or sham-vaccinated animals cleared the nasal infection with similar kinetics within 7 weeks, administration of the aP vaccine promoted persistent colonization of mouse nasal mucosa by B. pertussis.

ACS Style

Jana Holubová; Ondřej Staněk; Ludmila Brázdilová; Jiří Mašín; Ladislav Bumba; Andrew R. Gorringe; Frances Alexander; Peter Šebo. Acellular Pertussis Vaccine Inhibits Bordetella pertussis Clearance from the Nasal Mucosa of Mice. Vaccines 2020, 8, 695 .

AMA Style

Jana Holubová, Ondřej Staněk, Ludmila Brázdilová, Jiří Mašín, Ladislav Bumba, Andrew R. Gorringe, Frances Alexander, Peter Šebo. Acellular Pertussis Vaccine Inhibits Bordetella pertussis Clearance from the Nasal Mucosa of Mice. Vaccines. 2020; 8 (4):695.

Chicago/Turabian Style

Jana Holubová; Ondřej Staněk; Ludmila Brázdilová; Jiří Mašín; Ladislav Bumba; Andrew R. Gorringe; Frances Alexander; Peter Šebo. 2020. "Acellular Pertussis Vaccine Inhibits Bordetella pertussis Clearance from the Nasal Mucosa of Mice." Vaccines 8, no. 4: 695.

Journal article
Published: 15 September 2020 in Toxins
Reads 0
Downloads 0

Pathogenic Bordetella bacteria release a neurotropic dermonecrotic toxin (DNT) that is endocytosed into animal cells and permanently activates the Rho family GTPases by polyamination or deamidation of the glutamine residues in their switch II regions (e.g., Gln63 of RhoA). DNT was found to enable high level colonization of the nasal cavity of pigs by B. bronchiseptica and the capacity of DNT to inhibit differentiation of nasal turbinate bone osteoblasts causes atrophic rhinitis in infected pigs. However, it remains unknown whether DNT plays any role also in virulence of the human pathogen B. pertussis and in pathogenesis of the whooping cough disease. We report a procedure for purification of large amounts of LPS-free recombinant DNT that exhibits a high biological activity on cells expressing the DNT receptors Cav3.1 and Cav3.2Electron microscopy and single particle image analysis of negatively stained preparations revealed that the DNT molecule adopts a V-shaped structure with well-resolved protein domains. These results open the way to structure–function studies on DNT and its interactions with airway epithelial layers.

ACS Style

Ondrej Stanek; Irena Linhartova; Jana Holubova; Ladislav Bumba; Zdenko Gardian; Anna Malandra; Barbora Bockova; Shihono Teruya; Yasuhiko Horiguchi; Radim Osicka; Peter Sebo. Production of Highly Active Recombinant Dermonecrotic Toxin of Bordetella Pertussis. Toxins 2020, 12, 596 .

AMA Style

Ondrej Stanek, Irena Linhartova, Jana Holubova, Ladislav Bumba, Zdenko Gardian, Anna Malandra, Barbora Bockova, Shihono Teruya, Yasuhiko Horiguchi, Radim Osicka, Peter Sebo. Production of Highly Active Recombinant Dermonecrotic Toxin of Bordetella Pertussis. Toxins. 2020; 12 (9):596.

Chicago/Turabian Style

Ondrej Stanek; Irena Linhartova; Jana Holubova; Ladislav Bumba; Zdenko Gardian; Anna Malandra; Barbora Bockova; Shihono Teruya; Yasuhiko Horiguchi; Radim Osicka; Peter Sebo. 2020. "Production of Highly Active Recombinant Dermonecrotic Toxin of Bordetella Pertussis." Toxins 12, no. 9: 596.

Immunology
Published: 11 September 2020 in Frontiers in Immunology
Reads 0
Downloads 0

Circulating inflammatory monocytes are attracted to infected mucosa and differentiate into macrophage or dendritic cells endowed with enhanced bactericidal and antigen presenting capacities. In this brief Perspective we discuss the newly emerging insight into how the cAMP signaling capacity of Bordetella pertussis adenylate cyclase toxin manipulates the differentiation of monocytes and trigger dedifferentiation of the alveolar macrophages to facilitate bacterial colonization of human airways.

ACS Style

Jawid Nazir Ahmad; Peter Sebo. Adenylate Cyclase Toxin Tinkering With Monocyte-Macrophage Differentiation. Frontiers in Immunology 2020, 11, 1 .

AMA Style

Jawid Nazir Ahmad, Peter Sebo. Adenylate Cyclase Toxin Tinkering With Monocyte-Macrophage Differentiation. Frontiers in Immunology. 2020; 11 ():1.

Chicago/Turabian Style

Jawid Nazir Ahmad; Peter Sebo. 2020. "Adenylate Cyclase Toxin Tinkering With Monocyte-Macrophage Differentiation." Frontiers in Immunology 11, no. : 1.

Research article
Published: 10 August 2020 in PLOS Pathogens
Reads 0
Downloads 0

Bordetella bronchiseptica and Bordetella pertussis are closely related respiratory pathogens that evolved from a common bacterial ancestor. While B. bronchiseptica has an environmental reservoir and mostly establishes chronic infections in a broad range of mammals, B. pertussis is a human-specific pathogen causing acute pulmonary pertussis in infants and whooping cough illness in older humans. Both species employ a type III secretion system (T3SS) to inject a cytotoxic BteA effector protein into host cells. However, compared to the high BteA-mediated cytotoxicity of B. bronchiseptica, the cytotoxicity induced by B. pertussis BteA (Bp BteA) appears to be quite low and this has been attributed to the reduced T3SS gene expression in B. pertussis. We show that the presence of an alanine residue inserted at position 503 (A503) of Bp BteA accounts for its strongly attenuated cytotoxic potency. The deletion of A503 from Bp BteA greatly enhanced the cytotoxic activity of B. pertussis B1917 on mammalian HeLa cells and expression of Bp BteAΔA503 was highly toxic to Saccharomyces cerevisiae cells. Vice versa, insertion of A503 into B. bronchiseptica BteA (Bb BteA) strongly decreased its cytotoxicity to yeast and HeLa cells. Moreover, the production of Bp BteAΔA503 increased virulence of B. pertussis B1917 in the mouse model of intranasal infection (reduced LD50) but yielded less inflammatory pathology in infected mouse lungs at sublethal infectious doses. This suggests that A503 insertion in the T3SS effector Bp BteA may represent an evolutionary adaptation that fine-tunes B. pertussis virulence and host immune response. Pertussis remains the least-controlled vaccine-preventable infectious disease and the mechanisms by which Bordetella pertussis subverts defense mechanisms of human airway mucosa remain poorly understood. We found that B. pertussis had the cytotoxic activity of its type III secretion system-delivered effector BteA strongly attenuated by insertion of an alanine residue at position 503 as compared to the BteA homologue of the animal pathogen B. bronchiseptica. This functional adaptation reduced the capacity of B. pertussis to suppress host inflammatory response and may contribute to an acute course of the pulmonary form of human infant pertussis.

ACS Style

Jan Bayram; Ivana Malcova; Larisa Sinkovec; Jana Holubova; Gaia Streparola; David Jurnecka; Jan Kucera; Radislav Sedlacek; Peter Sebo; Jana Kamanova. Cytotoxicity of the effector protein BteA was attenuated in Bordetella pertussis by insertion of an alanine residue. PLOS Pathogens 2020, 16, e1008512 .

AMA Style

Jan Bayram, Ivana Malcova, Larisa Sinkovec, Jana Holubova, Gaia Streparola, David Jurnecka, Jan Kucera, Radislav Sedlacek, Peter Sebo, Jana Kamanova. Cytotoxicity of the effector protein BteA was attenuated in Bordetella pertussis by insertion of an alanine residue. PLOS Pathogens. 2020; 16 (8):e1008512.

Chicago/Turabian Style

Jan Bayram; Ivana Malcova; Larisa Sinkovec; Jana Holubova; Gaia Streparola; David Jurnecka; Jan Kucera; Radislav Sedlacek; Peter Sebo; Jana Kamanova. 2020. "Cytotoxicity of the effector protein BteA was attenuated in Bordetella pertussis by insertion of an alanine residue." PLOS Pathogens 16, no. 8: e1008512.

Journal article
Published: 07 August 2020 in The Journal of Immunology
Reads 0
Downloads 0

Despite high vaccine coverage in many parts of the world, pertussis is resurging in a number of areas in which acellular vaccines are the primary vaccine administered to infants and young children. This is attributed in part to the suboptimal and short-lived immunity elicited by acellular pertussis vaccines and to their inability to prevent nasal colonization and transmission of the etiologic agent Bordetella pertussis. In response to this escalating public health concern, the National Institute of Allergy and Infectious Diseases held the workshop “Overcoming Waning Immunity in Pertussis Vaccines” in September 2019 to identify issues and possible solutions for the defects in immunity stimulated by acellular pertussis vaccines. Discussions covered aspects of the current problem, gaps in knowledge and possible paths forward. This review summarizes presentations and discussions of some of the key points that were raised by the workshop.

ACS Style

F. Heath Damron; Mariette Barbier; Purnima Dubey; Kathryn M. Edwards; Xin-Xing Gu; Nicola P. Klein; Kristina Lu; Kingston H. G. Mills; Marcela F. Pasetti; Robert C. Read; Pejman Rohani; Peter Sebo; Eric T. Harvill. Overcoming Waning Immunity in Pertussis Vaccines: Workshop of the National Institute of Allergy and Infectious Diseases. The Journal of Immunology 2020, 205, 877 -882.

AMA Style

F. Heath Damron, Mariette Barbier, Purnima Dubey, Kathryn M. Edwards, Xin-Xing Gu, Nicola P. Klein, Kristina Lu, Kingston H. G. Mills, Marcela F. Pasetti, Robert C. Read, Pejman Rohani, Peter Sebo, Eric T. Harvill. Overcoming Waning Immunity in Pertussis Vaccines: Workshop of the National Institute of Allergy and Infectious Diseases. The Journal of Immunology. 2020; 205 (4):877-882.

Chicago/Turabian Style

F. Heath Damron; Mariette Barbier; Purnima Dubey; Kathryn M. Edwards; Xin-Xing Gu; Nicola P. Klein; Kristina Lu; Kingston H. G. Mills; Marcela F. Pasetti; Robert C. Read; Pejman Rohani; Peter Sebo; Eric T. Harvill. 2020. "Overcoming Waning Immunity in Pertussis Vaccines: Workshop of the National Institute of Allergy and Infectious Diseases." The Journal of Immunology 205, no. 4: 877-882.

Research article
Published: 16 July 2020 in Journal of Proteome Research
Reads 0
Downloads 0

Post-translational modifications of proteins enable swift physiological adaptation of cells to altered growth conditions and stress. Aside from protein phosphorylation, acetylation on ε-amino groups of lysine residues (N-ε-lysine acetylation) represents another important post-translational modification of proteins. For many bacterial pathogens, including the whooping cough agent Bordetella pertussis, the role and extent of protein acetylation remains to be defined. We expressed in E. coli the BP0960 and BP3063 genes encoding two putative deacetylases of B. pertussis and show that BP0960 encodes a lysine deacetylase enzyme, named Bkd1, that regulates acetylation of a range of B. pertussis proteins. Comparison of the proteome and acetylome of a bkd1 mutant with the proteome and acetylome of wild-type B. pertussis (PRIDE ID. PXD016384) revealed that acetylation on lysine residues may modulate activities or stabilities of proteins involved in bacterial metabolism and histone-like proteins. However, increased acetylation of the BvgA response regulator protein of the B. pertussis master virulence-regulating BvgAS two-component system affected neither the total levels of produced BvgA, nor its phosphorylation status. Indeed, the bkd1 mutant was not impaired in production of key virulence factors and its survival within human macrophages in vitro was not affected. The bkd1 mutant exhibited an increased growth rate under carbon source-limiting conditions and its virulence in the in vivo mouse lung infection model was somewhat affected. These results indicate that the lysine deacetylase Bkd1 and the N-ε-lysine acetylation primarily modulate the general metabolism rather than virulence of B. pertussis.

ACS Style

Jakub Novak; Ivo Fabrik; David Jurnečka; Jana Holubova; Ondrej Stanek; Peter Sebo. Bordetella pertussis Acetylome is Shaped by Lysine Deacetylase Bkd1. Journal of Proteome Research 2020, 19, 1 .

AMA Style

Jakub Novak, Ivo Fabrik, David Jurnečka, Jana Holubova, Ondrej Stanek, Peter Sebo. Bordetella pertussis Acetylome is Shaped by Lysine Deacetylase Bkd1. Journal of Proteome Research. 2020; 19 (9):1.

Chicago/Turabian Style

Jakub Novak; Ivo Fabrik; David Jurnečka; Jana Holubova; Ondrej Stanek; Peter Sebo. 2020. "Bordetella pertussis Acetylome is Shaped by Lysine Deacetylase Bkd1." Journal of Proteome Research 19, no. 9: 1.

Journal article
Published: 01 July 2020 in Journal of Biological Chemistry
Reads 0
Downloads 0

In a wide range of organisms, from bacteria to humans, numerous proteins have to be posttranslationally acylated to become biologically active. Bacterial repeats in toxin (RTX) cytolysins form a prominent group of proteins that are synthesized as inactive protoxins and undergo posttranslational acylation on ε-amino groups of two internal conserved lysine residues by co-expressed toxin-activating acyltransferases. Here, we investigated how the chemical nature, position, and number of bound acyl chains govern the activities of Bordetella pertussis adenylate cyclase toxin (CyaA), Escherichia coli α-hemolysin (HlyA), and Kingella kingae cytotoxin (RtxA). We found that the three protoxins are acylated in the same E. coli cell background by each of the CyaC, HlyC, and RtxC acyltransferases. We also noted that the acyltransferase selects from the bacterial pool of acyl–acyl carrier proteins (ACPs) an acyl chain of a specific length for covalent linkage to the protoxin. The acyltransferase also selects whether both or only one of two conserved lysine residues of the protoxin will be posttranslationally acylated. Functional assays revealed that RtxA has to be modified by 14-carbon fatty acyl chains to be biologically active, that HlyA remains active also when modified by 16-carbon acyl chains, and that CyaA is activated exclusively by 16-carbon acyl chains. These results suggest that the RTX toxin molecules are structurally adapted to the length of the acyl chains used for modification of their acylated lysine residue in the second, more conserved acylation site.

ACS Style

Adriana Osickova; Humaira Khaliq; Jiri Masin; David Jurnecka; Anna Sukova; Radovan Fiser; Jana Holubova; Ondrej Stanek; Peter Sebo; Radim Osicka. Acyltransferase-mediated selection of the length of the fatty acyl chain and of the acylation site governs activation of bacterial RTX toxins. Journal of Biological Chemistry 2020, 295, 9268 -9280.

AMA Style

Adriana Osickova, Humaira Khaliq, Jiri Masin, David Jurnecka, Anna Sukova, Radovan Fiser, Jana Holubova, Ondrej Stanek, Peter Sebo, Radim Osicka. Acyltransferase-mediated selection of the length of the fatty acyl chain and of the acylation site governs activation of bacterial RTX toxins. Journal of Biological Chemistry. 2020; 295 (28):9268-9280.

Chicago/Turabian Style

Adriana Osickova; Humaira Khaliq; Jiri Masin; David Jurnecka; Anna Sukova; Radovan Fiser; Jana Holubova; Ondrej Stanek; Peter Sebo; Radim Osicka. 2020. "Acyltransferase-mediated selection of the length of the fatty acyl chain and of the acylation site governs activation of bacterial RTX toxins." Journal of Biological Chemistry 295, no. 28: 9268-9280.

Journal article
Published: 01 July 2020 in Journal of Biological Chemistry
Reads 0
Downloads 0

The Bordetella adenylate cyclase toxin-hemolysin (CyaA) and the α-hemolysin (HlyA) of Escherichia coli belong to the family of cytolytic pore-forming Repeats in ToXin (RTX) cytotoxins. HlyA preferentially binds the αLβ2 integrin LFA-1 (CD11a/CD18) of leukocytes and can promiscuously bind and permeabilize also many other cells. CyaA bears an N-terminal adenylyl cyclase (AC) domain linked to a pore-forming RTX cytolysin (Hly) moiety, binds the complement receptor 3 (CR3, αMβ2, CD11b/CD18, or Mac-1) of myeloid phagocytes, penetrates their plasma membrane, and delivers the AC enzyme into the cytosol. We constructed a set of CyaA/HlyA chimeras and show that the CyaC-acylated segment and the CR3-binding RTX domain of CyaA can be functionally replaced by the HlyC-acylated segment and the much shorter RTX domain of HlyA. Instead of binding CR3, a CyaA1-710/HlyA411-1024 chimera bound the LFA-1 receptor and effectively delivered AC into Jurkat T cells. At high chimera concentrations (25 nM), the interaction with LFA-1 was not required for CyaA1-710/HlyA411-1024 binding to CHO cells. However, interaction with the LFA-1 receptor strongly enhanced the specific capacity of the bound CyaA1-710/HlyA411-1024 chimera to penetrate cells and deliver the AC enzyme into their cytosol. Hence, interaction of the acylated segment and/or the RTX domain of HlyA with LFA-1 promoted a productive membrane interaction of the chimera. These results help delimit residues 400–710 of CyaA as an “AC translocon” sufficient for translocation of the AC polypeptide across the plasma membrane of target cells.

ACS Style

Jiri Masin; Adriana Osickova; David Jurnecka; Nela Klimova; Humaira Khaliq; Peter Sebo; Radim Osicka. Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzyme–translocating segment of Bordetella adenylate cyclase toxin. Journal of Biological Chemistry 2020, 295, 9349 -9365.

AMA Style

Jiri Masin, Adriana Osickova, David Jurnecka, Nela Klimova, Humaira Khaliq, Peter Sebo, Radim Osicka. Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzyme–translocating segment of Bordetella adenylate cyclase toxin. Journal of Biological Chemistry. 2020; 295 (28):9349-9365.

Chicago/Turabian Style

Jiri Masin; Adriana Osickova; David Jurnecka; Nela Klimova; Humaira Khaliq; Peter Sebo; Radim Osicka. 2020. "Retargeting from the CR3 to the LFA-1 receptor uncovers the adenylyl cyclase enzyme–translocating segment of Bordetella adenylate cyclase toxin." Journal of Biological Chemistry 295, no. 28: 9349-9365.

Preprint content
Published: 03 April 2020
Reads 0
Downloads 0

Bordetella bronchisepticaandBordetella pertussisare closely related respiratory pathogens that evolved from a common bacterial ancestor. WhileB. bronchisepticahas an environmental reservoir and mostly establishes chronic infections in a broad range of mammals,B. pertussisis a human-specific pathogen causing acute pulmonary pertussis in infants and whooping cough illness in older humans. Both species employ a type III secretion system (T3SS) to inject a cytotoxic BteA effector protein into host cells. However, compared to the high BteA-mediated cytotoxicity ofB. bronchiseptica, the cytotoxicity induced byB. pertussisBteA (BpBteA) appears to be quite low and this has been attributed to the reduced T3SS gene expression inB. pertussis. We show that presence of an alanine residue inserted at position 503 (A503) ofBpBteA accounts for its strongly attenuated cytotoxic potency. Deletion of A503 fromBpBteA greatly enhanced the cytotoxic activity ofB. pertussisB1917 on mammalian HeLa cells and expression ofBpBteAΔA503 was highly toxic toSaccharomyces cerevisiaecells.Vice versa, insertion of A503 intoB. bronchisepticaBteA (BbBteA) strongly decreased its cytotoxicity to yeast and HeLa cells. Moreover, production ofBpBteAΔA503 increased virulence ofB. pertussisB1917 in the mouse model of intranasal infection (reduced LD50) but yielded less inflammatory pathology in infected mouse lungs at sublethal infectious doses. This suggests that A503 insertion in the T3SS effectorBpBteA may represent an evolutionary adaptation that fine-tunesB. pertussisvirulence and host immune response. Author summary Pertussis remains the least-controlled vaccine-preventable infectious disease and the mechanisms by whichBordetella pertussissubverts defense mechanisms of human airway mucosa remain poorly understood. We found thatB. pertussishad the cytotoxic activity of its type III secretion system-delivered effector BteA strongly attenuated by insertion of an alanine residue at position 503 as compared to the BteA homologue of the animal pathogenB. bronchiseptica. This functional adaptation reduced the capacity ofB. pertussisto suppress host inflammatory response and may contribute to an acute course of the pulmonary form of human infant pertussis.

ACS Style

Jan Bayram; Ivana Malcova; Larisa Sinkovec; Jana Holubova; Gaia Streparola; David Jurnecka; Jan Kucera; Radislav Sedlacek; Peter Sebo; Jana Kamanova. Cytotoxicity of the effector protein BteA was attenuated in Bordetella pertussis by insertion of an alanine residue. 2020, 1 .

AMA Style

Jan Bayram, Ivana Malcova, Larisa Sinkovec, Jana Holubova, Gaia Streparola, David Jurnecka, Jan Kucera, Radislav Sedlacek, Peter Sebo, Jana Kamanova. Cytotoxicity of the effector protein BteA was attenuated in Bordetella pertussis by insertion of an alanine residue. . 2020; ():1.

Chicago/Turabian Style

Jan Bayram; Ivana Malcova; Larisa Sinkovec; Jana Holubova; Gaia Streparola; David Jurnecka; Jan Kucera; Radislav Sedlacek; Peter Sebo; Jana Kamanova. 2020. "Cytotoxicity of the effector protein BteA was attenuated in Bordetella pertussis by insertion of an alanine residue." , no. : 1.

Journal article
Published: 29 October 2019 in mBio
Reads 0
Downloads 0

Macrophages are key sentinel cells of the immune system, and, as such, they are targeted by the toxins produced by the pertussis agent Bordetella pertussis . The adenylate cyclase toxin (CyaA) mediates immune evasion of B. pertussis by suspending the bactericidal activities of myeloid phagocytes. We reveal a novel mechanism of potential subversion of host immunity, where CyaA at very low (22 pM) concentrations could inhibit maturation of human monocyte precursors into the more phagocytic macrophage cells. Furthermore, exposure to low CyaA amounts has been shown to trigger dedifferentiation of mature primary human alveolar macrophages back into monocyte-like cells. This unprecedented capacity is likely to promote survival of the pathogen in the airways, both by preventing maturation of monocytes attracted to the site of infection into phagocytic macrophages and by dedifferentiation of the already airway-resident sentinel cells.

ACS Style

Jawid Nazir Ahmad; Jana Holubova; Oldrich Benada; Olga Kofronova; Ludek Stehlik; Martina Vasakova; Peter Sebo. Bordetella Adenylate Cyclase Toxin Inhibits Monocyte-to-Macrophage Transition and Dedifferentiates Human Alveolar Macrophages into Monocyte-like Cells. mBio 2019, 10, e01743-19 .

AMA Style

Jawid Nazir Ahmad, Jana Holubova, Oldrich Benada, Olga Kofronova, Ludek Stehlik, Martina Vasakova, Peter Sebo. Bordetella Adenylate Cyclase Toxin Inhibits Monocyte-to-Macrophage Transition and Dedifferentiates Human Alveolar Macrophages into Monocyte-like Cells. mBio. 2019; 10 (5):e01743-19.

Chicago/Turabian Style

Jawid Nazir Ahmad; Jana Holubova; Oldrich Benada; Olga Kofronova; Ludek Stehlik; Martina Vasakova; Peter Sebo. 2019. "Bordetella Adenylate Cyclase Toxin Inhibits Monocyte-to-Macrophage Transition and Dedifferentiates Human Alveolar Macrophages into Monocyte-like Cells." mBio 10, no. 5: e01743-19.

Journal article
Published: 20 June 2019 in Toxins
Reads 0
Downloads 0

Myeloid phagocytes have evolved to rapidly recognize invading pathogens and clear them through opsonophagocytic killing. The adenylate cyclase toxin (CyaA) of Bordetella pertussis and the edema toxin (ET) of Bacillus anthracis are both calmodulin-activated toxins with adenylyl cyclase activity that invade host cells and massively increase the cellular concentrations of a key second messenger molecule, 3’,5’-cyclic adenosine monophosphate (cAMP). However, the two toxins differ in the kinetics and mode of cell entry and generate different cAMP concentration gradients within the cell. While CyaA rapidly penetrates cells directly across their plasma membrane, the cellular entry of ET depends on receptor-mediated endocytosis and translocation of the enzymatic subunit across the endosomal membrane. We show that CyaA-generated membrane-proximal cAMP gradient strongly inhibits the activation and phosphorylation of Syk, Vav, and Pyk2, thus inhibiting opsonophagocytosis. By contrast, at similar overall cellular cAMP levels, the ET-generated perinuclear cAMP gradient poorly inhibits the activation and phosphorylation of these signaling proteins. Hence, differences in spatiotemporal distribution of cAMP produced by the two adenylyl cyclase toxins differentially affect the opsonophagocytic signaling in myeloid phagocytes.

ACS Style

Shakir Hasan; Waheed Ur Rahman; Peter Sebo; Radim Osicka. Distinct Spatiotemporal Distribution of Bacterial Toxin-Produced Cellular cAMP Differentially Inhibits Opsonophagocytic Signaling. Toxins 2019, 11, 362 .

AMA Style

Shakir Hasan, Waheed Ur Rahman, Peter Sebo, Radim Osicka. Distinct Spatiotemporal Distribution of Bacterial Toxin-Produced Cellular cAMP Differentially Inhibits Opsonophagocytic Signaling. Toxins. 2019; 11 (6):362.

Chicago/Turabian Style

Shakir Hasan; Waheed Ur Rahman; Peter Sebo; Radim Osicka. 2019. "Distinct Spatiotemporal Distribution of Bacterial Toxin-Produced Cellular cAMP Differentially Inhibits Opsonophagocytic Signaling." Toxins 11, no. 6: 362.

Journal article
Published: 12 June 2019 in Toxins
Reads 0
Downloads 0

Cytolytic leukotoxins of the repeat in toxin (RTX) family are large proteins excreted by gram-negative bacterial pathogens through the type 1 secretion system (T1SS). Due to low yields and poor stability in cultures of the original pathogens, it is useful to purify recombinant fatty-acylated RTX cytolysins from inclusion bodies produced in E. coli. Such preparations are, however, typically contaminated by high amounts of E. coli lipopolysaccharide (LPS or endotoxin). We report a simple procedure for purification of large amounts of biologically active and endotoxin-free RTX toxins. It is based on the common feature of RTX cytolysins that are T1SS-excreted as unfolded polypeptides and fold into a biologically active toxin only upon binding of calcium ions outside of the bacterial cell. Mimicking this process, the RTX proteins are solubilized from inclusion bodies with buffered 8 M urea, bound onto a suitable chromatographic medium under denaturing conditions and the contaminating LPS is removed through extensive on-column washes with buffers containing 6 to 8 M urea and 1% Triton X-100 or Triton X-114. Extensive on-column rinsing with 8 M urea buffer removes residual detergent and the eluted highly active RTX protein preparations then contain only trace amounts of LPS. The procedure is exemplified using four prototypic RTX cytolysins, the Bordetella pertussis CyaA and the hemolysins of Escherichia coli (HlyA), Kingella kingae (RtxA), and Actinobacillus pleuropneumoniae (ApxIA).

ACS Style

Ondrej Stanek; Jiri Masin; Radim Osicka; David Jurnecka; Adriana Osickova; Peter Sebo. Rapid Purification of Endotoxin-Free RTX Toxins. Toxins 2019, 11, 336 .

AMA Style

Ondrej Stanek, Jiri Masin, Radim Osicka, David Jurnecka, Adriana Osickova, Peter Sebo. Rapid Purification of Endotoxin-Free RTX Toxins. Toxins. 2019; 11 (6):336.

Chicago/Turabian Style

Ondrej Stanek; Jiri Masin; Radim Osicka; David Jurnecka; Adriana Osickova; Peter Sebo. 2019. "Rapid Purification of Endotoxin-Free RTX Toxins." Toxins 11, no. 6: 336.

Journal article
Published: 08 April 2019 in Scientific Reports
Reads 0
Downloads 0

The adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) of pathogenic Bordetellae delivers its adenylyl cyclase (AC) enzyme domain into the cytosol of host cells and catalyzes uncontrolled conversion of cellular ATP to cAMP. In parallel, the toxin forms small cation-selective pores that permeabilize target cell membrane and account for the hemolytic activity of CyaA on erythrocytes. The pore-forming domain of CyaA is predicted to consist of five transmembrane α-helices, of which the helices I, III, IV and V have previously been characterized. We examined here the α-helix II that is predicted to form between residues 529 to 549. Substitution of the glycine 531 residue by a proline selectively reduced the hemolytic capacity but did not affect the AC translocating activity of the CyaA-G531P toxin. In contrast, CyaA toxins with alanine 538 or 546 replaced by diverse residues were selectively impaired in the capacity to translocate the AC domain across cell membrane but remained fully hemolytic. Such toxins, however, formed pores in planar asolectin bilayer membranes with a very low frequency and with at least two different conducting states. The helix-breaking substitution of alanine 538 by a proline residue abolished the voltage-activated increase of membrane activity of CyaA in asolectin bilayers. These results reveal that the predicted α-helix comprising the residues 529 to 549 plays a key role in CyaA penetration into the target plasma membrane and pore-forming activity of the toxin.

ACS Style

Jana Roderova; Adriana Osickova; Anna Sukova; Gabriela Mikusova; Radovan Fiser; Peter Sebo; Radim Osicka; Jiri Masin. Residues 529 to 549 participate in membrane penetration and pore-forming activity of the Bordetella adenylate cyclase toxin. Scientific Reports 2019, 9, 5758 .

AMA Style

Jana Roderova, Adriana Osickova, Anna Sukova, Gabriela Mikusova, Radovan Fiser, Peter Sebo, Radim Osicka, Jiri Masin. Residues 529 to 549 participate in membrane penetration and pore-forming activity of the Bordetella adenylate cyclase toxin. Scientific Reports. 2019; 9 (1):5758.

Chicago/Turabian Style

Jana Roderova; Adriana Osickova; Anna Sukova; Gabriela Mikusova; Radovan Fiser; Peter Sebo; Radim Osicka; Jiri Masin. 2019. "Residues 529 to 549 participate in membrane penetration and pore-forming activity of the Bordetella adenylate cyclase toxin." Scientific Reports 9, no. 1: 5758.

Personal view
Published: 28 November 2018 in The Lancet Infectious Diseases
Reads 0
Downloads 0

Summary The resurgence and changing epidemiology of pertussis in high-income countries, the high infant mortality caused by pertussis in low-income countries, and the increasing morbidity in all age groups worldwide call for a concerted effort to both improve the current vaccines and develop new vaccines and vaccination strategies against pertussis. In this Personal View, we identify several key obstacles on the path to developing a durable solution for global control of pertussis. To systematically address these obstacles, the PERtussIS Correlates Of Protection Europe (PERISCOPE) Consortium was established in March, 2016. The objectives of this consortium are to increase scientific understanding of immunity to pertussis in humans induced by vaccines and infections, to identify biomarkers of protective immunity, and to generate technologies and infrastructure for the future development of improved pertussis vaccines. By working towards the accelerated licensure and implementation of novel, well tolerated, and effective pertussis vaccines, we hope to strengthen and stimulate further collaboration and transparency between the key stakeholders, including the public, the scientific community, public health institutes, regulatory authorities, and vaccine manufacturers.

ACS Style

Dimitri A. Diavatopoulos; Kingston H.G. Mills; Kent E. Kester; Beate Kampmann; Marcela Silerova; Ulrich Heininger; Jacques J.M. van Dongen; Robbert G. van der Most; Martijn A. Huijnen; Emilio Siena; Nathalie Mielcarek; Martina M. Ochs; Philippe Denoël; Guy Berbers; Annemarie M. Buisman; Marien I. de Jonge; Craig Fenwick; Andrew Gorringe; Qiushui He; Dominic Kelly; Roger Le Grand; Camille Locht; Françoise Mascart; Jussi Mertsola; Alberto Orfao; Giuseppe Pantaleo; Andrew J. Pollard; Andrew Preston; Robert Read; Peter Sebo; Cecile van Els; Branislav Vecerek; Patricia Londoño-Hayes; Ronald de Groot. PERISCOPE: road towards effective control of pertussis. The Lancet Infectious Diseases 2018, 19, e179 -e186.

AMA Style

Dimitri A. Diavatopoulos, Kingston H.G. Mills, Kent E. Kester, Beate Kampmann, Marcela Silerova, Ulrich Heininger, Jacques J.M. van Dongen, Robbert G. van der Most, Martijn A. Huijnen, Emilio Siena, Nathalie Mielcarek, Martina M. Ochs, Philippe Denoël, Guy Berbers, Annemarie M. Buisman, Marien I. de Jonge, Craig Fenwick, Andrew Gorringe, Qiushui He, Dominic Kelly, Roger Le Grand, Camille Locht, Françoise Mascart, Jussi Mertsola, Alberto Orfao, Giuseppe Pantaleo, Andrew J. Pollard, Andrew Preston, Robert Read, Peter Sebo, Cecile van Els, Branislav Vecerek, Patricia Londoño-Hayes, Ronald de Groot. PERISCOPE: road towards effective control of pertussis. The Lancet Infectious Diseases. 2018; 19 (5):e179-e186.

Chicago/Turabian Style

Dimitri A. Diavatopoulos; Kingston H.G. Mills; Kent E. Kester; Beate Kampmann; Marcela Silerova; Ulrich Heininger; Jacques J.M. van Dongen; Robbert G. van der Most; Martijn A. Huijnen; Emilio Siena; Nathalie Mielcarek; Martina M. Ochs; Philippe Denoël; Guy Berbers; Annemarie M. Buisman; Marien I. de Jonge; Craig Fenwick; Andrew Gorringe; Qiushui He; Dominic Kelly; Roger Le Grand; Camille Locht; Françoise Mascart; Jussi Mertsola; Alberto Orfao; Giuseppe Pantaleo; Andrew J. Pollard; Andrew Preston; Robert Read; Peter Sebo; Cecile van Els; Branislav Vecerek; Patricia Londoño-Hayes; Ronald de Groot. 2018. "PERISCOPE: road towards effective control of pertussis." The Lancet Infectious Diseases 19, no. 5: e179-e186.

Journal article
Published: 03 September 2018 in Pathogens and Disease
Reads 0
Downloads 0

Bordetella pertussis is a strictly human pathogen causing the respiratory infectious disease called whooping cough or pertussis. B. pertussis adaptation to acellular pertussis vaccine pressure has been repeatedly highlighted, but recent data indicate that adaptation of circulating strains started already in the era of the whole cell pertussis vaccine (wP) use. We sequenced the genomes of five B. pertussis wP vaccine strains isolated in the former Czechoslovakia in the pre-wP (1954–1957) and early wP (1958–1965) eras, when only limited population travel into and out of the country was possible. Four isolates exhibit a similar genome organization and form a distinct phylogenetic cluster with a geographic signature. The fifth strain is rather distinct, both in genome organization and SNP-based phylogeny. Surprisingly, despite isolation of this strain before 1966, its closest sequenced relative appears to be a recent isolate from the US. On the genome content level, the five vaccine strains contained both new and already described regions of difference. One of the new regions contains duplicated genes potentially associated with transport across the membrane. The prevalence of this region in recent isolates indicates that its spread might be associated with selective advantage leading to increased strain fitness.

ACS Style

Ana Dienstbier; Derek Pouchnik; Mark Wildung; Fabian Amman; Ivo L Hofacker; Julian Parkhill; Jana Holubova; Peter Sebo; Branislav Vecerek. Comparative genomics of Czech vaccine strains of Bordetella pertussis. Pathogens and Disease 2018, 76, 1 .

AMA Style

Ana Dienstbier, Derek Pouchnik, Mark Wildung, Fabian Amman, Ivo L Hofacker, Julian Parkhill, Jana Holubova, Peter Sebo, Branislav Vecerek. Comparative genomics of Czech vaccine strains of Bordetella pertussis. Pathogens and Disease. 2018; 76 (7):1.

Chicago/Turabian Style

Ana Dienstbier; Derek Pouchnik; Mark Wildung; Fabian Amman; Ivo L Hofacker; Julian Parkhill; Jana Holubova; Peter Sebo; Branislav Vecerek. 2018. "Comparative genomics of Czech vaccine strains of Bordetella pertussis." Pathogens and Disease 76, no. 7: 1.

Journal article
Published: 16 June 2018 in Toxins
Reads 0
Downloads 0

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.

ACS Style

Ladislav Bumba; Jiri Masin; Adriana Osickova; Radim Osicka; Peter Sebo. 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. Toxins 2018, 10, 245 .

AMA Style

Ladislav Bumba, Jiri Masin, Adriana Osickova, Radim Osicka, Peter Sebo. 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. Toxins. 2018; 10 (6):245.

Chicago/Turabian Style

Ladislav Bumba; Jiri Masin; Adriana Osickova; Radim Osicka; Peter Sebo. 2018. "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." Toxins 10, no. 6: 245.