A Detailed Guide for Metabolic Incorporation of N-acetyl muramic Acid Probes into Bacterial Peptidoglycan
Bacterial cells utilize small carbohydrate building blocks to construct peptidoglycan (PG), a highly conserved mesh-like polymer that serves as a protective coat for the cell. PG production has long been a target for antibiotics, and its breakdown is a source for human immune recognition. A key comp...
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| Autores principales: | , , , , , , |
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| Formato: | Articulo Preprint |
| Lenguaje: | Inglés |
| Publicado: |
2019
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| Materias: | |
| Acceso en línea: | http://sedici.unlp.edu.ar/handle/10915/127285 https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpch.74 |
| Aporte de: |
| id |
I19-R120-10915-127285 |
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| record_format |
dspace |
| institution |
Universidad Nacional de La Plata |
| institution_str |
I-19 |
| repository_str |
R-120 |
| collection |
SEDICI (UNLP) |
| language |
Inglés |
| topic |
Ciencias Exactas Química carbohydrates bacterial peptidoglycan metabolic incorporation mass spectrometry microscopy fluorescent labeling bioorthogonal chemistry click chemistry |
| spellingShingle |
Ciencias Exactas Química carbohydrates bacterial peptidoglycan metabolic incorporation mass spectrometry microscopy fluorescent labeling bioorthogonal chemistry click chemistry DeMeester, Kristen E. Liang, Hai Zhou, Junhui Wodzanowski, Kimberly A. Prather, Benjamin L. Santiago, Cintia C. Grimes, Catherine L. A Detailed Guide for Metabolic Incorporation of N-acetyl muramic Acid Probes into Bacterial Peptidoglycan |
| topic_facet |
Ciencias Exactas Química carbohydrates bacterial peptidoglycan metabolic incorporation mass spectrometry microscopy fluorescent labeling bioorthogonal chemistry click chemistry |
| description |
Bacterial cells utilize small carbohydrate building blocks to construct peptidoglycan (PG), a highly conserved mesh-like polymer that serves as a protective coat for the cell. PG production has long been a target for antibiotics, and its breakdown is a source for human immune recognition. A key component of bacterial PG, N-acetyl muramic acid (NAM), is a vital element in many synthetically derived immunostimulatory compounds. However, the exact molecular details of these structures and how they are generated remain unknown due to a lack of chemical probes surrounding the NAM core. A robust synthetic strategy to generate bioorthogonally tagged NAM carbohydrate units is implemented. These molecules serve as precursors for PG biosynthesis and recycling. Escherichia coli cells are metabolically engineered to incorporate the bioorthogonal NAM probes into their PG network. The probes are subsequently modified using copper-catalyzed azide-alkyne cycloaddition to install fluorophores directly into the bacterial PG, as confirmed by super-resolution microscopy and high-resolution mass spectrometry. Here, synthetic notes for key elements of this process to generate the sugar probes as well as streamlined user-friendly metabolic labeling strategies for both microbiology and immunological applications are described. |
| format |
Articulo Preprint |
| author |
DeMeester, Kristen E. Liang, Hai Zhou, Junhui Wodzanowski, Kimberly A. Prather, Benjamin L. Santiago, Cintia C. Grimes, Catherine L. |
| author_facet |
DeMeester, Kristen E. Liang, Hai Zhou, Junhui Wodzanowski, Kimberly A. Prather, Benjamin L. Santiago, Cintia C. Grimes, Catherine L. |
| author_sort |
DeMeester, Kristen E. |
| title |
A Detailed Guide for Metabolic Incorporation of N-acetyl muramic Acid Probes into Bacterial Peptidoglycan |
| title_short |
A Detailed Guide for Metabolic Incorporation of N-acetyl muramic Acid Probes into Bacterial Peptidoglycan |
| title_full |
A Detailed Guide for Metabolic Incorporation of N-acetyl muramic Acid Probes into Bacterial Peptidoglycan |
| title_fullStr |
A Detailed Guide for Metabolic Incorporation of N-acetyl muramic Acid Probes into Bacterial Peptidoglycan |
| title_full_unstemmed |
A Detailed Guide for Metabolic Incorporation of N-acetyl muramic Acid Probes into Bacterial Peptidoglycan |
| title_sort |
detailed guide for metabolic incorporation of n-acetyl muramic acid probes into bacterial peptidoglycan |
| publishDate |
2019 |
| url |
http://sedici.unlp.edu.ar/handle/10915/127285 https://currentprotocols.onlinelibrary.wiley.com/doi/10.1002/cpch.74 |
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Repositorios |
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