Open AccessProtected N-Acetyl Muramic Acid Probes Improve Bacterial Peptidoglycan Incorporation via Metabolic Labeling
Author(s) -
Ashley R. Brown,
Kimberly A. Wodzanowski,
Cintia C. Santiago,
Stephen N. Hyland,
Julianna L Follmar,
PapaNii Asare-Okai,
Catherine L. Grimes
Publication year - 2021
Publication title -
acs chemical biology
Language(s) - Uncategorized
Resource type - Journals
SCImago Journal Rank - 1.899
H-Index - 111
eISSN - 1554-8937
pISSN - 1554-8929
DOI - 10.1021/acschembio.1c00268
Subject(s) - peptidoglycan , glycan , bacterial cell structure , biochemistry , cell envelope , cell wall , glycobiology , muramic acid , biology , bacteria , computational biology , chemistry , escherichia coli , glycoprotein , gene , genetics
Metabolic glycan probes have emerged as an excellent tool to investigate vital questions in biology. Recently, methodology to incorporate metabolic bacterial glycan probes into the cell wall of a variety of bacterial species has been developed. In order to improve this method, a scalable synthesis of the peptidoglycan precursors is developed here, allowing for access to essential peptidoglycan immunological fragments and cell wall building blocks. The question was asked if masking polar groups of the glycan probe would increase overall incorporation, a common strategy exploited in mammalian glycobiology. Here, we show, through cellular assays, that E. coli do not utilize peracetylated peptidoglycan substrates but do employ methyl esters. The 10-fold improvement of probe utilization indicates that (i) masking the carboxylic acid is favorable for transport and (ii) bacterial esterases are capable of removing the methyl ester for use in peptidoglycan biosynthesis. This investigation advances bacterial cell wall biology, offering a prescription on how to best deliver and utilize bacterial metabolic glycan probes.