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Tausch, F.* ; Dietrich, R.* ; Schauer, K.* ; Janowski, R. ; Niessing, D. ; Märtlbauer, E.* ; Jessberger, N.*

Evidence for complex formation of the Bacillus cereus haemolysin BL components in solution.

Toxins 9:288 (2017)
Verlagsversion Forschungsdaten DOI PMC
Open Access Gold
Creative Commons Lizenzvertrag
Haemolysin BL is an important virulence factor regarding the diarrheal type of food poisoning caused by Bacillus cereus. However, the pathogenic importance of this three-component enterotoxin is difficult to access, as nearly all natural B. cereus culture supernatants additionally contain the highly cytotoxic Nhe, the second three-component toxin involved in the aetiology of B. cereus-induced food-borne diseases. To better address the toxic properties of the Hbl complex, a system for overexpression and purification of functional, cytotoxic, recombinant (r)Hbl components L₂, L₁ and B from E. coli was established and an nheABC deletion mutant was constructed from B. cereus reference strain F837/76. Furthermore, 35 hybridoma cell lines producing monoclonal antibodies (mAbs) against Hbl L₂, L₁ and B were generated. While mAbs 1H9 and 1D8 neutralized Hbl toxicity and thus, represent important tools for future investigations of the mode-of-action of Hbl on the target cell surface, mAb 1D7, in contrast, even enhanced Hbl toxicity by supporting the binding of Hbl B to the cell surface. By using the specific mAbs in Dot blots, indirect and hybrid sandwich enzyme immuno assays (EIAs), complex formation between Hbl L₁ and B, as well as L₁ and L₂ in solution could be shown for the first time. Surface plasmon resonance experiments with the rHbl components confirmed these results with KD values of 4.7 × 10(-7) M and 1.5 × 10(-7) M, respectively. These findings together with the newly created tools lay the foundation for the detailed elucidation of the molecular mode-of-action of the highly complex three-component Hbl toxin.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter Bacillus Cereus ; Complex Formation ; Enterotoxins ; Haemolysin Bl ; Monoclonal Antibodies; Antibody-dependent Enhancement; Nonhemolytic Enterotoxin Complex; 3 Exoprotein Components; Monoclonal-antibodies; Nhe Enterotoxin; Virus-infection; Toxin; Cytotoxicity; Virulence; Family
e-ISSN 2072-6651
Zeitschrift Toxins
Quellenangaben Band: 9, Heft: 9, Seiten: , Artikelnummer: 288 Supplement: ,
Verlag MDPI
Verlagsort Basel
Nichtpatentliteratur Publikationen
Begutachtungsstatus Peer reviewed
Institut(e) Institute of Structural Biology (STB)