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Title 

Shiga toxin A subunit mutant of Escherichia coli O157:H7 releases outer membrane vesicles containing the B-pentameric complex

Authors 

Sang-Hyun KimSang Rae LeeK S KimA KoEkyune KimY H KimKyu Tae Chang

Publisher 

Wiley-Blackwell

Issue Date 

2010

Citation 

FEMS Immunology and Medical Microbiology, vol. 58, no. 3, pp. 412-420

Abstract 

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.

ISSN 

0928-8244

Link 

http://dx.doi.org/10.1111/j.1574-695X.2010.00654.x

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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