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Mechanisms of incorporation for D-amino acid probes that target peptidoglycan biosynthesis
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Kuru, Erkin, Radkov, Atanas, Meng, Xin, Egan, Alexander, Alvarez, Laura, Dowson, Amanda, Booher, Garrett, Breukink, Eefjan, Roper, David I., Cava, Felipe, Vollmer, Waldemar, Brun, Yves and VanNieuwenhze, Michael S. (2019) Mechanisms of incorporation for D-amino acid probes that target peptidoglycan biosynthesis. ACS Chemical Biology, 14 (12). pp. 2745-2756. doi:10.1021/acschembio.9b00664 ISSN 1554-8929.
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WRAP-mechanisms-incorporation-acid-probes-Roper-2019.pdf - Accepted Version Embargoed item. Restricted access to Repository staff only - Requires a PDF viewer. Download (2504Kb) |
Official URL: http://dx.doi.org/10.1021/acschembio.9b00664
Abstract
Bacteria exhibit a myriad of different morphologies, through the synthesis and modification of their essential peptidoglycan (PG) cell wall. Our discovery of a fluorescent D-amino acid (FDAA)-based PG labeling approach provided a powerful method for observing how these morphological changes occur. Given that PG is unique to bacterial cells and a common target for antibiotics, understanding the precise mechanism(s) for incorporation of (F)DAA-based probes is a crucial determinant in understanding the role of PG synthesis in bacterial cell biology and could provide a valuable tool in the development of new antimicrobials to treat drug-resistant antibacterial infections. Here, we systematically investigate the mechanisms of FDAA probe incorporation into PG using two model organisms Escherichia coli (Gram-negative) and Bacillus subtilis (Gram-positive). Our in vitro and in vivo data unequivocally demonstrate that these bacteria incorporate FDAAs using two extracytoplasmic pathways: through activity of their D,D-transpeptidases, and, if present, by their L,D-transpeptidases and not via cytoplasmic incorporation into a D-Ala-D-Ala dipeptide precursor. Our data also revealed the unprecedented finding that the DAA-drug, D-cycloserine, can be incorporated into peptide stems by each of these transpeptidases, in addition to its known inhibitory activity against D-alanine racemase and D-Ala-D-Ala ligase. These mechanistic findings enabled development of a new, FDAA-based, in vitro labeling approach that reports on subcellular distribution of muropeptides, an especially important attribute to enable study of bacteria with poorly defined growth modes. An improved understanding of the incorporation mechanisms utilized by DAA-based probes is essential when interpreting results from high resolution experiments and highlights the antimicrobial potential of synthetic DAAs.
Item Type: | Journal Article | |||||||||||||||||||||
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Subjects: | Q Science > QD Chemistry Q Science > QP Physiology Q Science > QR Microbiology |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | |||||||||||||||||||||
Library of Congress Subject Headings (LCSH): | Amino acids, Peptidoglycans, Bacterial cell walls, Viruses -- Morphology | |||||||||||||||||||||
Journal or Publication Title: | ACS Chemical Biology | |||||||||||||||||||||
Publisher: | American Chemical Society | |||||||||||||||||||||
ISSN: | 1554-8929 | |||||||||||||||||||||
Official Date: | 20 December 2019 | |||||||||||||||||||||
Dates: |
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Volume: | 14 | |||||||||||||||||||||
Number: | 12 | |||||||||||||||||||||
Page Range: | pp. 2745-2756 | |||||||||||||||||||||
DOI: | 10.1021/acschembio.9b00664 | |||||||||||||||||||||
Status: | Peer Reviewed | |||||||||||||||||||||
Publication Status: | Published | |||||||||||||||||||||
Reuse Statement (publisher, data, author rights): | This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Chemical Biology, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acschembio.9b00664 | |||||||||||||||||||||
Access rights to Published version: | Restricted or Subscription Access | |||||||||||||||||||||
Copyright Holders: | © American Chemical Society | |||||||||||||||||||||
Date of first compliant deposit: | 28 November 2019 | |||||||||||||||||||||
Date of first compliant Open Access: | 7 January 2020 | |||||||||||||||||||||
RIOXX Funder/Project Grant: |
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