Production of Highly Active Recombinant Dermonecrotic Toxin ofBordetella Pertussis

0535246 - MBÚ 2021 RIV CH eng J - Journal Article
Staněk, Ondřej - Linhartová, Irena - Holubová, Jana - Bumba, Ladislav - Gardian, Zdenko - Malandra, Anna - Bočková, Barabora - Teruya, S. - Horiguchi, Y. - Osička, Radim - Šebo, Peter
Production of Highly Active Recombinant Dermonecrotic Toxin ofBordetella Pertussis.
Toxins. Roč. 12, č. 9 (2020), č. článku 596. ISSN 2072-6651. E-ISSN 2072-6651
R&D Projects: GA ČR(CZ) GX19-27630X; GA ČR(CZ) GA19-12695S; GA MŠMT(CZ) LM2018133
Research Infrastructure: Czech-BioImaging - 90062
Institutional support: RVO:61388971 ; RVO:60077344
Keywords : Bordetella * GTPase * deamidation * dermonecrotic toxin * recombinant
OECD category: Microbiology; Biochemical research methods (BC-A)
Impact factor: 4.546, year: 2020
Method of publishing: Open access
https://www.mdpi.com/2072-6651/12/9/596

PathogenicBordetellabacteria 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 byB. bronchisepticaand 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 pathogenB. pertussisand 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.
Permanent Link: http://hdl.handle.net/11104/0313335