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Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays

Peptidoglycan (PG) is an essential cell wall component that maintains the morphology and viability of nearly all bacteria. Its biosynthesis requires periplasmic transpeptidation reactions which construct peptide cross-linkages between polysaccharide chains to endow mechanical strength. However, trac...

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Autores principales: Hsu, Yen-Pang, Hall, Edward, Booher, Garrett, Murphy, Brennan, Radkov, Atanas D., Yablonowski, Jacob, Mulcahey, Caitlyn, Alvarez, Laura, Cava, Felipe, Brun, Yves V., Kuru, Erkin, VanNieuwenhze, Michael S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6444347/
https://www.ncbi.nlm.nih.gov/pubmed/30804500
http://dx.doi.org/10.1038/s41557-019-0217-x
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author Hsu, Yen-Pang
Hall, Edward
Booher, Garrett
Murphy, Brennan
Radkov, Atanas D.
Yablonowski, Jacob
Mulcahey, Caitlyn
Alvarez, Laura
Cava, Felipe
Brun, Yves V.
Kuru, Erkin
VanNieuwenhze, Michael S.
author_facet Hsu, Yen-Pang
Hall, Edward
Booher, Garrett
Murphy, Brennan
Radkov, Atanas D.
Yablonowski, Jacob
Mulcahey, Caitlyn
Alvarez, Laura
Cava, Felipe
Brun, Yves V.
Kuru, Erkin
VanNieuwenhze, Michael S.
author_sort Hsu, Yen-Pang
collection PubMed
description Peptidoglycan (PG) is an essential cell wall component that maintains the morphology and viability of nearly all bacteria. Its biosynthesis requires periplasmic transpeptidation reactions which construct peptide cross-linkages between polysaccharide chains to endow mechanical strength. However, tracking transpeptidation reaction in vivo and in vitro is challenging, mainly due to the lack of efficient, biocompatible probes. Here, we report the design, synthesis, and application of rotor-fluorogenic D-amino acids (RfDAAs) enabling real-time, continuous tracking of transpeptidation reactions. These probes enable monitoring PG biosynthesis in real time through visualizing transpeptidase reactions in live cells, as well as real-time activity assays of D,D-, L,D-transpeptidases, and sortases in vitro. The unique ability of RfDAAs to become fluorescent when incorporated into PG provides a powerful new tool to study PG biosynthesis with high temporal resolution and prospectively enable high-throughput screening for inhibitors of PG biosynthesis.
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spelling pubmed-64443472019-08-25 Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays Hsu, Yen-Pang Hall, Edward Booher, Garrett Murphy, Brennan Radkov, Atanas D. Yablonowski, Jacob Mulcahey, Caitlyn Alvarez, Laura Cava, Felipe Brun, Yves V. Kuru, Erkin VanNieuwenhze, Michael S. Nat Chem Article Peptidoglycan (PG) is an essential cell wall component that maintains the morphology and viability of nearly all bacteria. Its biosynthesis requires periplasmic transpeptidation reactions which construct peptide cross-linkages between polysaccharide chains to endow mechanical strength. However, tracking transpeptidation reaction in vivo and in vitro is challenging, mainly due to the lack of efficient, biocompatible probes. Here, we report the design, synthesis, and application of rotor-fluorogenic D-amino acids (RfDAAs) enabling real-time, continuous tracking of transpeptidation reactions. These probes enable monitoring PG biosynthesis in real time through visualizing transpeptidase reactions in live cells, as well as real-time activity assays of D,D-, L,D-transpeptidases, and sortases in vitro. The unique ability of RfDAAs to become fluorescent when incorporated into PG provides a powerful new tool to study PG biosynthesis with high temporal resolution and prospectively enable high-throughput screening for inhibitors of PG biosynthesis. 2019-02-25 2019-04 /pmc/articles/PMC6444347/ /pubmed/30804500 http://dx.doi.org/10.1038/s41557-019-0217-x Text en Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use:http://www.nature.com/authors/editorial_policies/license.html#terms
spellingShingle Article
Hsu, Yen-Pang
Hall, Edward
Booher, Garrett
Murphy, Brennan
Radkov, Atanas D.
Yablonowski, Jacob
Mulcahey, Caitlyn
Alvarez, Laura
Cava, Felipe
Brun, Yves V.
Kuru, Erkin
VanNieuwenhze, Michael S.
Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays
title Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays
title_full Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays
title_fullStr Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays
title_full_unstemmed Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays
title_short Fluorogenic D-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays
title_sort fluorogenic d-amino acids enable real-time monitoring of peptidoglycan biosynthesis and high-throughput transpeptidation assays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6444347/
https://www.ncbi.nlm.nih.gov/pubmed/30804500
http://dx.doi.org/10.1038/s41557-019-0217-x
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