The DOI matches its Crossref record; 43 of the 43 references it cites were found in a registry or an index.
Synthesis of fluorescent D-amino acids and their use for probing peptidoglycan synthesis and bacterial growth in situ
Open accessAbstract
Fluorescent D-amino acids (FDAAs) are efficiently incorporated into the peptidoglycans (PGs) of diverse bacterial species at the sites of PG biosynthesis, allowing specific and covalent probing of bacterial growth with minimal perturbation. Here we provide a protocol for the synthesis of four FDAAs emitting light in blue (HCC-amino-D-alanine, HADA), green (NBD-amino-D-alanine, NADA, and fluorescein-D-lysine, FDL) or red (TAMRA-D-lysine, TDL) and for their use in PG labeling of live bacteria. Our modular synthesis protocol gives easy access to a library of different FDAAs made with commercially available fluorophores and diamino acid starting materials. Molecules can be synthesized in a typical chemistry laboratory in 2–3 d using standard chemical transformations. The simple labeling procedure involves the addition of the FDAAs to a bacterial sample for the desired labeling duration and stopping further label incorporation by fixing the cells with cold 70% (vol/vol) ethanol or by washing away excess dye. We discuss several scenarios for the use of these labels in fluorescence microscopy applications, including short or long labeling durations, and the combination of different labels in pure culture (e.g., for 'virtual time-lapse' microscopy) or in situ labeling of complex environmental samples. Depending on the experiment, FDAA labeling can take as little as 30 s for a rapidly growing species such as Escherichia coli.
Overview
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Citations
Cites
The crisis in antibiotic resistance
Neu, H. C. (1992) · Science · doi:10.1126/science.257.5073.1064 · Cited by 2,834 · cited 1×
“HADA (3) and NADA (5), which are currently the most robust DAA derivatives, can be prepared in good yields from N- -Boc–protected 3-aminod-alanine (1), via nucleophilic capture of an activated ester of 7-hydroxycoumarin-3-carboxylic acid (HCC-OH; 2) or by nucleophilic aromatic substitution of 4-chloro-7-nitrobenzo2-ox…”
Peptidoglycan structure and architecture
Vollmer, W., Blanot, D., & De Pedro, M. A. (2008) · FEMS Microbiology Reviews · doi:10.1111/j.1574-6976.2007.00094.x · Cited by 2,340
Prescher, J. A., & Bertozzi, C. R. (2005) · Nature Chemical Biology · doi:10.1038/nchembio0605-13 · Cited by 1,456
Growth of the Stress-Bearing and Shape-Maintaining Murein Sacculus of Escherichia coli
Höltje, J.-V. (1998) · Microbiology and Molecular Biology Reviews · doi:10.1128/mmbr.62.1.181-203.1998 · Cited by 1,129
Control of Cell Morphogenesis in Bacteria
Daniel, R. A., & Errington, J. (2003) · Cell · doi:10.1016/s0092-8674(03)00421-5 · Cited by 740
In Situ Probing of Newly Synthesized Peptidoglycan in Live Bacteria with Fluorescent D‐Amino Acids
Kuru, E., Hughes, H. V., Brown, P. J., Hall, E., Tekkam, S., Cava, F., de Pedro, M. A., Brun, Y. V., & VanNieuwenhze, M. S. (2012) · Angewandte Chemie International Edition · doi:10.1002/anie.201206749 · Cited by 627
Self-citationD-Amino Acids Govern Stationary Phase Cell Wall Remodeling in Bacteria
Lam, H., Oh, D.-C., Cava, F., Takacs, C. N., Clardy, J., de Pedro, M. A., & Waldor, M. K. (2009) · Science · doi:10.1126/science.1178123 · Cited by 618
Emerging knowledge of regulatory roles of d-amino acids in bacteria
Cava, F., Lam, H., de Pedro, M. A., & Waldor, M. K. (2011) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-010-0571-8 · Cited by 400
Murein segregation in Escherichia coli
de Pedro, M. A., Quintela, J. C., Höltje, J. V., & Schwarz, H. (1997) · Journal of Bacteriology · doi:10.1128/jb.179.9.2823-2834.1997 · Cited by 388
A new metabolic cell-wall labelling method reveals peptidoglycan in Chlamydia trachomatis
Liechti, G. W., Kuru, E., Hall, E., Kalinda, A., Brun, Y. V., VanNieuwenhze, M., & Maurelli, A. T. (2014) · Nature · doi:10.1038/nature12892 · Cited by 354
Self-citation
Cited by
Treadmilling by FtsZ filaments drives peptidoglycan synthesis and bacterial cell division
Bisson-Filho, A. W., Hsu, Y.-P., Squyres, G. R., Kuru, E., Wu, F., Jukes, C., Sun, Y., Dekker, C., Holden, S., VanNieuwenhze, M. S., et al. (2017) · Science · doi:10.1126/science.aak9973 · Cited by 601
Self-citationThe mycobacterial cell envelope — a moving target
Dulberger, C. L., Rubin, E. J., & Boutte, C. C. (2020) · Nature Reviews Microbiology · doi:10.1038/s41579-019-0273-7 · Cited by 365
Maturing Mycobacterium smegmatis peptidoglycan requires non-canonical crosslinks to maintain shape
Baranowski, C., Welsh, M. A., Sham, L.-T., Eskandarian, H. A., Lim, H. C., Kieser, K. J., Wagner, J. C., McKinney, J. D., Fantner, G. E., Ioerger, T. R., et al. (2018) · eLife · doi:10.7554/elife.37516 · Cited by 132
Imaging Bacterial Cell Wall Biosynthesis
Radkov, A. D., Hsu, Y.-P., Booher, G., & VanNieuwenhze, M. S. (2018) · Annual Review of Biochemistry · doi:10.1146/annurev-biochem-062917-012921 · Cited by 102
Self-citationImaging mycobacterial growth and division with a fluorogenic probe
Hodges, H. L., Brown, R. A., Crooks, J. A., Weibel, D. B., & Kiessling, L. L. (2018) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1720996115 · Cited by 96
Metadata
- Authors
- Kuru, E., Tekkam, S., Hall, E., Brun, Y. V., & Van Nieuwenhze, M. S.
- Venue
- Nature Protocols
- Published
- 2014-12-04
- Volume
- 10
- Issue
- 1
- Pages
- 33-52
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W2080878746
- Cited by
- 310
- References
- 43
- FWCI
- 10.70
- Type
- journal article
- Language
- en
- Open access
- green
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
How to cite
Cite CoChat for the checks on this page, and the paper for its own findings.
- This page
- CoChat. (2026). Citation check: Synthesis of fluorescent D-amino acids and their use for probing peptidoglycan synthesis and bacterial growth in situ. https://app.cochat.ai/papers/10.1038/nprot.2014.197
- The paper
- Kuru, E., Tekkam, S., Hall, E., Brun, Y. V., & Van Nieuwenhze, M. S. (2015). Synthesis of fluorescent D-amino acids and their use for probing peptidoglycan synthesis and bacterial growth in situ. Nature Protocols. https://doi.org/10.1038/nprot.2014.197
Data sources
Bibliographic data: Semantic Scholar, OpenAlex, Crossref.
Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
