| Mouse toxicity and cytokine release by verotoxin 1 B subunit mutants. | |
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MedLine Citation:
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PMID: 11119557 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The crystal structure of the verotoxin 1 (VT1) B subunit complexed with a globotriaosylceramide (Gb(3)) analogue showed the presence of three receptor binding sites per monomer. We wished to study the effects of altering the three sites, singly or in combination, on animal toxicity and cytokine induction in vitro. We found that while the site 1 and 2 mutants were modestly (two- to sevenfold) reduced in their ability to cause disease in BALB/c mice, the site 3 mutant, W34A, was as toxic as VT1. However, all the double-mutant proteins, irrespective of which two sites were mutated, exhibited approximately a 100-fold reduction in their 50% lethal doses for mice. These results suggest that multivalent receptor binding is important in vivo and that all three binding sites make a similar contribution to the latter process. The triple-mutant holotoxin, F30A G62T W34A, administered intraperitoneally without adjuvant, stimulated a strong antibody response in BALB/c mice, and the immune sera neutralized the activity of VT1 in vitro. Induction of tumor neurosis factor alpha release from differentiated human monocytes (THP-1 cells) was relatively impaired for site 1 and site 2 but not site 3 mutants, suggesting an auxiliary role for the latter site in mediation of cytokine release in vitro. Cytotoxicity assays on undifferentiated THP-1 cells have also demonstrated the importance of sites 1 and 2 and the relatively small role played by site 3 in causing cell death. These data suggest an association between the cytotoxicity of the protein and its ability to induce cytokine release. |
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Authors:
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V M Wolski; A M Soltyk; J L Brunton |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Infection and immunity Volume: 69 ISSN: 0019-9567 ISO Abbreviation: Infect. Immun. Publication Date: 2001 Jan |
Date Detail:
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Created Date: 2001-01-18 Completed Date: 2001-01-18 Revised Date: 2013-04-17 |
Medline Journal Info:
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Nlm Unique ID: 0246127 Medline TA: Infect Immun Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 579-83 Citation Subset: IM |
Affiliation:
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Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Line Escherichia coli / pathogenicity* Female Glycosides / immunology Humans Mice Mice, Inbred BALB C Monocytes / drug effects, metabolism Mutation Protein Subunits Shiga Toxins / immunology, metabolism, toxicity* Trihexosylceramides / metabolism Triterpenes / immunology Tumor Necrosis Factor-alpha / biosynthesis* |
| Chemical | |
Reg. No./Substance:
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0/Glycosides; 0/Protein Subunits; 0/Shiga Toxins; 0/Trihexosylceramides; 0/Triterpenes; 0/Tumor Necrosis Factor-alpha; 37341-37-0/stichoposide; 71965-57-6/globotriaosylceramide |
| Comments/Corrections | |
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