Shiga toxin A subunit mutant of Escherichia coli O157:H7 releases outer membrane vesicles containing the B-pentameric complexopen access
- Authors
- Kim, Sang-Hyun; Lee, Sang-Rae; Kim, Keun-Su; Ko, Ara; Kim, Ekyune; Kim, Yong-Hwan; Chang, Kyu-Tae
- Issue Date
- Apr-2010
- Publisher
- WILEY
- Keywords
- outer membrane vesicle detoxification; median lethal dose; Shiga toxin A mutation; Escherichia coli O157; vaccine vehicle
- Citation
- FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY, v.58, no.3, pp.412 - 420
- Indexed
- SCIE
SCOPUS
- Journal Title
- FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY
- Volume
- 58
- Number
- 3
- Start Page
- 412
- End Page
- 420
- URI
- https://scholarworks.bwise.kr/gnu/handle/sw.gnu/25148
- DOI
- 10.1111/j.1574-695X.2010.00654.x
- ISSN
- 0928-8244
- Abstract
- Shiga toxins (STx) are secreted extracellularly through the outer membrane vesicles (OMVs) of Escherichia coli O157:H7. In an attempt to produce STxA-deficient OMVs from E. coli O157:H7, site-specific deletions of the stx1A and stx2A subunit genes were carried out. The STxA-deficient phenotype of the stx1A/stx2A mutant was confirmed by Vero cell cytotoxicity and VTEC-RPLA (R) assay. Western blot analyses showed that the B (STxB) subunits were present without coupling to STxA in the OMVs of the STxA-deficient mutant. Furthermore, STxB was located in its homo-pentameric complexes, as revealed by immunoprecipitation and immunoblotting with anti-STxB antibodies. These results suggest that STxB alone can be oligomerized into the B pentamer in the periplasm, and subsequently entrapped into the OMVs. Determination of the median lethal dose concentration for the OMV preparations suggests that the STxA-deficient OMVs containing STxB complex could be safely used as vaccine delivery vehicles.
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