| Site-specific incorporation of photocrosslinker and bioorthogonal amino acids into enteric bacterial pathogens. | |
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MedLine Citation:
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PMID: 22084898 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Enteric bacterial pathogens are known to effectively pass through the extremely acidic mammalian stomachs and cause infections in the small and/or large intestine of human hosts. However, their acid-survival strategy and pathogenesis mechanisms remain elusive, largely due to the lack of tools to directly monitor and manipulate essential components (e.g. defense proteins or invasive toxins) participating these processes. Herein, we have extended the pyrrolysine-based genetic-code expansion strategy for encoding unnatural amino acids (UAAs) in enteric bacterial species including enteropathogenic E. coli (EPEC), Shigella and Salmonella. Using this system, a photocrosslinking amino acid was incorporated into a Shigella acid-chaperone HdeA (shHdeA), which allowed the identification of a comprehensive list of in vivo client proteins that are protected by shHdeA upon acid stress. To further demonstrate the application of our strategy, an azide bearing amino acid was introduced into a Shigella Type Three Secretion (T3S) effector-OspF without interrupting its secretion efficiency. This site-specifically installed azide-handle allowed the facile detection of OspF's secretion in bacterial extracellular space. Taken together, these bioorthogonal functionalities we incorporated into enteric pathogens were shown to facilitate the investigation of unique and important proteins involved in the pathogenesis and stress-defense mechanisms of pathogenic bacteria that remain exceedingly difficult to study using conventional methodologies. |
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Authors:
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Shixian Lin; Zhenrun Zhang; Hao Xu; Lin Li; She Chen; Jie Li; Ziyang Hao; Peng R Chen |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-11-15 |
Journal Detail:
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Title: Journal of the American Chemical Society Volume: - ISSN: 1520-5126 ISO Abbreviation: - Publication Date: 2011 Nov |
Date Detail:
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Created Date: 2011-11-16 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 7503056 Medline TA: J Am Chem Soc Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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