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Site-Specific Incorporation of Photo-Cross-Linker and Bioorthogonal Amino Acids into Enteric Bacterial Pathogens
Lin, Shixian1; Zhang, Zhenrun1; Xu, Hao3; Li, Lin3; Chen, She3; Li, Jie1; Hao, Ziyang1; Chen, Peng R.1,2
2011-12-21
Source PublicationJOURNAL OF THE AMERICAN CHEMICAL SOCIETY
ISSN0002-7863
Volume133Issue:50Pages:20581-20587
AbstractEnteric 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 in these processes. Herein, we have extended the pyrrolysine-based genetic code expansion strategy for encoding unnatural amino acids in enteric bacterial species, including enteropathogenic Escherichia coli, Shigella, and Salmonella. Using this system, a photo-cross-linking 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 3 secretion effector, OspF, without interruption of 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.
DOI10.1021/ja209008w
Indexed BySCI
Language英语
WOS IDWOS:000298713600081
PublisherAMER CHEMICAL SOC
Citation statistics
Cited Times:47[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.iccas.ac.cn/handle/121111/73770
Collection中国科学院化学研究所
Corresponding AuthorChen, Peng R.
Affiliation1.Peking Univ, Coll Chem & Mol Engn, Beijing Natl Lab Mol Sci, Beijing 100871, Peoples R China
2.Ctr Life Sci, Beijing 100871, Peoples R China
3.Natl Inst Biol Sci NIBS, Beijing 102206, Peoples R China
Recommended Citation
GB/T 7714
Lin, Shixian,Zhang, Zhenrun,Xu, Hao,et al. Site-Specific Incorporation of Photo-Cross-Linker and Bioorthogonal Amino Acids into Enteric Bacterial Pathogens[J]. JOURNAL OF THE AMERICAN CHEMICAL SOCIETY,2011,133(50):20581-20587.
APA Lin, Shixian.,Zhang, Zhenrun.,Xu, Hao.,Li, Lin.,Chen, She.,...&Chen, Peng R..(2011).Site-Specific Incorporation of Photo-Cross-Linker and Bioorthogonal Amino Acids into Enteric Bacterial Pathogens.JOURNAL OF THE AMERICAN CHEMICAL SOCIETY,133(50),20581-20587.
MLA Lin, Shixian,et al."Site-Specific Incorporation of Photo-Cross-Linker and Bioorthogonal Amino Acids into Enteric Bacterial Pathogens".JOURNAL OF THE AMERICAN CHEMICAL SOCIETY 133.50(2011):20581-20587.
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