Synthesis and secretion of Bordetella pertussis adenylate cyclase as a 200-kilodalton protein

Bellalou, Jacques; Ladant, Daniel; Sakamoto, Hiroshi

doi:10.1128/iai.58.5.1195-1200.1990

Cited by 40 publications

(12 citation statements)

References 25 publications

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“…However, when SS supernates of 48 h were concentrated, it was possible to detect the band of 200 kDa by immunoblotting (data not shown). This result is in agreement with that reported by Bellalou et al [23], who showed that various Bordetella species secrete the high molecular mass form of AC (200 kDa, the major intracellular form) into the super- nates of SS medium without proteolytic processing. In addition to enhancing the level of the intracellular form of AC (200 kDa) in supernates, it has been reported that cyclodextrin also enhanced the levels of two other extracellular proteins, PT and FHA [9,10].…”

Section: Resultssupporting

confidence: 93%

Use of cyclodextrin as an agent to induce excretion of Bordetella pertussis antigens

Hozbor

1994

FEMS Immunology and Medical Microbiology

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This paper attempts to provide an explanation for the effect of cyclodextrin on the yield of Bordetella pertussis soluble antigens. It was demonstrated that the addition of cyclodextrin to the synthetic Stainer-Scholte liquid medium enhances the level of the intracellular form of adenylate cyclase (200 kDa) in the supernate. In addition to this effect, it has been reported that cyclodextrin also enhances the levels of two other extracellular proteins, pertussis toxin and filamentous hemagglutinin. As these antigens are structurally different, it seems that the effect of cyclodextrin is not specific. With the use of different buffer systems of well-known action on outer membrane stability it was possible to determine a relationship between the presence of cyclodextrin, destabilisation of the outer membrane and the release of proteins. It was determined that the cyclodextrin did not modify the fluidity of B. pertussis cells but produced a change of outer membrane permeability.

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Section: Resultssupporting

confidence: 93%

Use of cyclodextrin as an agent to induce excretion of Bordetella pertussis antigens

Hozbor

1994

FEMS Immunology and Medical Microbiology

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“…including Escherichia coli a-hemolysin. Both B. pertussis and B. parapertussis synthesize a very similar AC-Hly toxin (5,14,16), although some differences in molecular weight and enzyme activities were observed (16). To be active, AC-WHy toxin requires posttranslational modification mediated by the protein coded for by the cyaC gene (4,32).…”

Section: Resultsmentioning

confidence: 99%

Bordetella pertussis and Bordetella parapertussis: two immunologically distinct species

et al. 1993

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Bordetella pertussis and Bordetella parapertussis are closely related species. Both are responsible for outbreaks of whooping cough in humans and produce similar virulence factors, with the exception of pertussis toxin, specific to B. pertussis. Current pertussis whole-cell vaccine will soon be replaced by acellular vaccines containing major adhesins (filamentous hemagglutinin and pertactin) and major toxin (pertussis toxin). All of these factors are antigens that stimulate a protective immune response in the murine respiratory model and in clinical assays. In the present study, we examined the protective efficacies of these factors, and that of adenylate cyclase-hemolysin, another B. pertussis toxin, against B. parapertussis infection in a murine respiratory model. As expected, pertussis toxin did not protect against B. parapertussis infection, since this bacterium did not express this protein, but the surprising result was that none of the other factors were protective against B. parapertussis infection. Furthermore, B. parapertussis adenylate cyclase-hemolysin, although it protected against B. parapertussis infection, did not protect against B. pertussis infection. Despite a high degree of homology between both B. pertussis and B. parapertussis species, no cross-protection was observed. Our results outline the fact that, as in other gram-negative bacteria, Bordetella surface proteins vary immunologically.

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“…The cytotoxic activity of CyaA is the result of an invasive and calmodulin-activated adenylate cyclase (AC) domain that penetrates across the cytoplasmic membrane of a variety of immune effector cells and impairs their physiological functions by catalysing uncontrolled conversion of ATP into cAMP (Confer and Eaton, 1982;Gordon et aL, 1989;Hanski and Farfel, 1985;Pearson et aL, 1987;Wolff et aL, 1980). In addition, this 180 kDa toxin is also endowed with haemolytic activity (Bellalou et aL, 1990a;Ehrmann • et aL, 1992;Gross et aL, 1992;Rogel et aL, 1991;Sakamoto et aL, 1992). This is the result of the ability of CyaA to form cation-selective channels that alter the permeability of target cell membranes and cause colloid-osmotic cell lysis (Benz et aL, 1994;Szabo et aL, 1994).…”

Section: Introductionmentioning

confidence: 99%

Identification by in vitro complementation of regions required for cell‐invasive activity of Bordetella pertussis adenylate cyclase toxin

Iwaki

Ullmann

Šebo

1995

Molecular Microbiology

102

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The adenylate cyclase toxin (CyaA) of Bordetella pertussis is a 1706-residue protein composed of an amino-terminal adenylate cyclase (AC) domain linked to a 1300-residue channel-forming RTX (repeats in toxin) haemolysin. The toxin delivers its AC domain into a variety of eukaryotic cells and impairs cellular functions by catalysing unregulated synthesis of cAMP from intracellular ATP. We have examined toxin activities of a set of deletion derivatives of CyaA. The results indicate that CyaA does not have a dedicated target cell-binding domain and that structural integrity and co-operation of all domains, as well as the post-translational fatty acylation mediated by an accessory protein CyaC, are all essential for target cell association and toxin activity of CyaA. When tested individually, all toxin derivatives were inactive and impaired in the tight association with the target cell surface. However, pairs of constructs with nonoverlapping deletions complemented each other in vitro and exhibited a partially restored cytotoxic activity. This suggests that at least a part of the active toxin may act in the form of dimers or higher oligomers. The complementation analysis revealed that the last 217 residues of CyaA, containing the unprocessed secretion signal, form an autonomous domain essential for toxin activity, and that the region from residue 624 to 780 may be directly involved in delivery of the AC toxin into cells.

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Synthesis and secretion of Bordetella pertussis adenylate cyclase as a 200-kilodalton protein

Cited by 40 publications

References 25 publications

Use of cyclodextrin as an agent to induce excretion of Bordetella pertussis antigens

Use of cyclodextrin as an agent to induce excretion of Bordetella pertussis antigens

Bordetella pertussis and Bordetella parapertussis: two immunologically distinct species

Identification by in vitro complementation of regions required for cell‐invasive activity of Bordetella pertussis adenylate cyclase toxin

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