Production of Highly Active Recombinant Dermonecrotic Toxin of Bordetella Pertussis

Toxins (Basel). 2020 Sep 15;12(9):596. doi: 10.3390/toxins12090596.


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 Cav3.2. Electron 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.

Keywords: Bordetella; GTPase; deamidation; dermonecrotic toxin; electron microscopy; image analysis; negative staining; recombinant.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • A549 Cells
  • Animals
  • Animals, Newborn
  • Bordetella pertussis / enzymology*
  • Bordetella pertussis / genetics
  • Bordetella pertussis / pathogenicity
  • Calcium Channels, T-Type / genetics
  • Calcium Channels, T-Type / metabolism
  • Epithelial Cells / metabolism*
  • Epithelial Cells / ultrastructure
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Necrosis
  • Protein Binding
  • Protein Domains
  • Recombinant Proteins / metabolism
  • Skin / drug effects
  • Skin / pathology
  • Structure-Activity Relationship
  • Transglutaminases / genetics
  • Transglutaminases / metabolism*
  • Transglutaminases / toxicity
  • Transglutaminases / ultrastructure
  • Virulence Factors, Bordetella / genetics
  • Virulence Factors, Bordetella / metabolism*
  • Virulence Factors, Bordetella / toxicity


  • CACNA1G protein, human
  • CACNA1H protein, human
  • Calcium Channels, T-Type
  • Recombinant Proteins
  • Virulence Factors, Bordetella
  • dermonecrotic toxin, Bordetella
  • Transglutaminases