Metabolic labeling of the bacterial peptidoglycan by functionalized glucosamine.
(2022) iScience — Vol. 25, n° 8, p. 104753 [1-19] (2022)
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- Abstract
- -Acetylglucosamine (GlcNAc) is an essential monosaccharide required in almost all organisms. Fluorescent labeling of the peptidoglycan (PG) on -acetylglucosamine has been poorly explored. Here, we report on the labeling of the PG with a bioorthogonal handle on the GlcNAc. We developed a facile one-step synthesis of uridine diphosphate -azidoacetylglucosamine (UDP-GlcNAz) using the glycosyltransferase OleD, followed by incorporation of GlcNAz into the peptidoglycan precursor Lipid II and fluorescent labeling of the azido group via click chemistry. In a PG synthesis assay, fluorescent GlcNAz-labeled Lipid II was incorporated into peptidoglycan by the DD-transpeptidase activity of bifunctional class A penicillin-binding proteins. We further demonstrate the incorporation of GlcNAz into the PG layer of OleD-expressed bacteria by feeding with 2-chloro-4-nitrophenyl GlcNAz (GlcNAz-CNP). Hence, our labeling method using the heterologous expression of OleD is useful to study PG synthesis and possibly other biological processes involving GlcNAc metabolism .
- Affiliations
Utrecht University Department of Chemistry
Citations
Xu, Y., Hernández-Rocamora, V. M., Lorent, J., Cox, R., Wang, X., Bao, X., Stel, M., Vos, G., van den Bos, R. M., Pieters, R. J., Gray, J., Vollmer, W., & Breukink, E. (2022). Metabolic labeling of the bacterial peptidoglycan by functionalized glucosamine. iScience, 25(8), 104753 [1-19]. https://doi.org/10.1016/j.isci.2022.104753 (Original work published 2022)