Uptake of unnatural trehalose analogs as a reporter for Mycobacterium tuberculosis
Open accessAbstract
The detection of tuberculosis currently relies upon insensitive and unspecific techniques; newer diagnostics would ideally co-opt specific bacterial processes to provide real-time readouts. The trehalose mycolyltransesterase enzymes (antigens 85A, 85B and 85C (Ag85A, Ag85B, Ag85C)) serve as essential mediators of cell envelope function and biogenesis in Mycobacterium tuberculosis. Through the construction of a systematically varied sugar library, we show here that Ag85 enzymes have exceptionally broad substrate specificity. This allowed exogenously added synthetic probes to be specifically incorporated into M. tuberculosis growing in vitro and within macrophages. Even bulky substituents, such as a fluorescein-containing trehalose probe (FITC-trehalose), were incorporated by growing bacilli, thereby producing fluorescent bacteria; microscopy revealed selective labeling of poles and membrane. Addition of FITC-trehalose to M. tuberculosis-infected macrophages allowed selective, sensitive detection of M. tuberculosis within infected mammalian macrophages. These studies suggest that analogs of trehalose may prove useful as probes of function and for other imaging modalities.
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Bioorthogonal metabolic labeling allows the biosynthesis of trehalose-containing molecules, peptidoglycans, and arabinan to be visualized in situ in both M. smegmatis and M. tuberculosis.
“Nevertheless, there are emerging efforts to apply modern (click chemistry) approaches of bioorthogonal metabolic labeling in both M. smegmatis and M. tuberculosis, allowing the biosynthesis of trehalose-containing molecules, peptidoglycans, and arabinan to be visualized in situ (116, 163–165).”
From Mycobacterium smegmatis: The Vanguard of Mycobacterial Research (2023) · cites this paper as [164]
The Results section ('FITC-trehalose specifically labels M. tuberculosis in vitro'; page unavailable) reports fluorescent visualization of trehalose-probe incorporation in live M. tuberculosis. This paper does not demonstrate the claimed labeling of peptidoglycan or arabinan, or the use of click chemistry; its mention of M. smegmatis is background rather than a demonstration of this probe in that species. (LLM, conf=0.86)
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- Authors
- Backus, K. M., Boshoff, H. I., Barry, C. S., Boutureira, O., Patel, M. K., D'Hooge, F., Lee, S. S., Via, L. E., Tahlan, K., Barry, C. E., & Davis, B. G.
- Affiliations
- University of Oxford (GB)
- National Institute of Allergy and Infectious Diseases (US)
- Corresponding author
- Barry, Conor S
- Barry, Clifton E
- Davis, Benjamin G
- ORCID
- Venue
- Nature Chemical Biology
- Published
- 2011-03-06
- Volume
- 7
- Issue
- 4
- Pages
- 228-235
- ISSN
- 1552-4450 (ISSN-L)
- 1552-4469
- PMC
- PMC3157484
- OpenAlex
- W2068962699
- Cited by
- 237
- References
- 46
- Citations by year
- 2026: 8 · 2025: 14 · 2024: 17 · 2023: 18 · 2022: 21 · 2021: 16 · 2020: 23 · 2019: 18 · 2018: 19 · 2017: 15 · 2016: 13 · 2015: 11 · 2014: 15 · 2013: 10 · 2012: 10
- FWCI
- 9.54
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Chemical Biology (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- ePrints Soton (University of Southampton) (acceptedVersion)
- Funders
- National Institutes of Health
- Engineering and Physical Sciences Research Council (EP/E000614/1, EP/D023343/1, GR/T26542/01, EP/I500200/1, EP/G026688/1, EP/D023335/1)
- Biotechnology and Biological Sciences Research Council (BB/C510824/1, BB/E004350/1, EGA17763)
- National Institute of Allergy and Infectious Diseases (Z01AI000693, ZIAAI000693, ZIAAI000734)
- Topics
- Health Sciences › Medicine › Infectious Diseases › Tuberculosis Research and Epidemiology
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Physical Sciences › Chemistry › Organic Chemistry › Carbohydrate Chemistry and Synthesis
- Keywords
- Mycobacterium tuberculosis, substrate specificity, fluorescent probes, bacterial cell wall, macrophage infection, tuberculosis diagnosis, bacterial imaging
- MeSH terms
- Animals, Cell Line, Fluorescent Dyes, Macrophages, Microscopy, Fluorescence, Mycobacterium tuberculosis, Mass Spectrometry, Substrate Specificity, Trehalose, Genes, Reporter, Mice
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