Production of Highly Active Recombinant Dermonecrotic Toxin of Bordetella Pertussis

Autor: Radim Osicka, Jana Holubova, Irena Linhartova, Yasuhiko Horiguchi, Peter Sebo, Ladislav Bumba, Zdenko Gardian, Ondrej Stanek, Anna Malandra, Shihono Teruya, Barbora Bockova
Jazyk: angličtina
Rok vydání: 2020
Předmět:
electron microscopy
negative staining
image analysis
Bordetella pertussis
RHOA
Bordetella
Health
Toxicology and Mutagenesis
lcsh:Medicine
GTPase
Toxicology
medicine.disease_cause
law.invention
Calcium Channels
T-Type
Mice
law
Virulence Factors
Bordetella
recombinant
Receptor
Skin
Mice
Inbred BALB C
0303 health sciences
biology
Chemistry
3T3 Cells
negative staining
Recombinant Proteins
deamidation
Recombinant DNA
Protein Binding
Virulence
dermonecrotic toxin
Article
Microbiology
Necrosis
Structure-Activity Relationship
03 medical and health sciences
Protein Domains
image analysis
medicine
Animals
Humans
030304 developmental biology
Transglutaminases
electron microscopy
030306 microbiology
Toxin
lcsh:R
Epithelial Cells
biology.organism_classification
Animals
Newborn
A549 Cells
biology.protein
Zdroj: Toxins, Vol 12, Iss 596, p 596 (2020)
Toxins
Volume 12
Issue 9
ISSN: 2072-6651
Popis: 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&ndash
function studies on DNT and its interactions with airway epithelial layers.
Databáze: OpenAIRE
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