d -Amino Acid Chemical Reporters Reveal Peptidoglycan Dynamics of an Intracellular Pathogen
Open accessAbstract
Peptidoglycan (PG) is an essential component of the bacterial cell wall. Although experiments with organisms in vitro have yielded a wealth of information on PG synthesis and maturation, it is unclear how these studies translate to bacteria replicating within host cells. We report a chemical approach for probing PG in vivo via metabolic labeling and bioorthogonal chemistry. A wide variety of bacterial species incorporated azide and alkyne-functionalized d-alanine into their cell walls, which we visualized by covalent reaction with click chemistry probes. The d-alanine analogues were specifically incorporated into nascent PG of the intracellular pathogen Listeria monocytogenes both in vitro and during macrophage infection. Metabolic incorporation of d-alanine derivatives and click chemistry detection constitute a facile, modular platform that facilitates unprecedented spatial and temporal resolution of PG dynamics in vivo.
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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 [116]
Figure 1 and the accompanying text (section labeled Abstract; page unavailable) show click-chemistry labeling of cell walls in M. tuberculosis using d-alanine reporters, supporting the peptidoglycan portion of the citation cluster. This source does not establish visualization of trehalose-containing molecules or arabinan, or labeling in M. smegmatis; its intracellular labeling experiments concern L. monocytogenes. (LLM, conf=0.87)
Experimental — accuracy not yet measuredD-alanine is used by bacteria to generate peptidoglycan but is not used by mammalian cells.
“Although D-alanine is used by bacteria to generate peptidoglycan, an essential component of the bacterial cell wall, it is not used by mammalian cells (fig. S14) (47).”
From The human tumor microbiome is composed of tumor type–specific intracellular bacteria (2020) · cites this paper as [47]
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Self-citation
- Authors
- Siegrist, M. S., Whiteside, S., Jewett, J. C., Aditham, A., Cava, F., & Bertozzi, C. R.
- Affiliations
- University of California, Berkeley (US)
- Howard Hughes Medical Institute (US)
- University of Arizona (US)
- Centro de Biología Molecular Severo Ochoa (ES)
- Corresponding author
- Bertozzi, Carolyn R.
- Venue
- ACS Chemical Biology
- Publisher
- American Chemical Society (ACS)
- Published
- 2013-01-11
- Volume
- 8
- Issue
- 3
- Pages
- 500-505
- ISSN
- 1554-8929 (ISSN-L)
- 1554-8937
- PMC
- PMC3601600
- OpenAlex
- W2019795913
- Cited by
- 246
- References
- 39
- Citations by year
- 2026: 7 · 2025: 11 · 2024: 22 · 2023: 16 · 2022: 15 · 2021: 25 · 2020: 23 · 2019: 17 · 2018: 18 · 2017: 23 · 2016: 15 · 2015: 20 · 2014: 23 · 2013: 11
- FWCI
- 7.95
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Locations
- ACS Chemical Biology (publishedVersion, other-oa)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- American Cancer Society
- Topics
- Physical Sciences › Chemistry › Organic Chemistry › Click Chemistry and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Structural Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
- Keywords
- Listeria monocytogenes, intracellular bacterial pathogens, metabolic labeling, bioorthogonal chemistry, macrophage infection, bacterial cell wall synthesis
- MeSH terms
- Alanine, Listeria monocytogenes, Molecular Conformation, Peptidoglycan, Molecular Dynamics Simulation, Click Chemistry
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