An OregonGreen488-labelled d-amino acid for visualizing peptidoglycan by super-resolution STED nanoscopy

Microbiology (Reading). 2020 Dec;166(12):1129-1135. doi: 10.1099/mic.0.000996.

Abstract

Fluorescent d-amino acids (FDAAs) are molecular probes that are widely used for labelling the peptidoglycan layer of bacteria. When added to growing cells they are incorporated into the stem peptide by a transpeptidase reaction, allowing the timing and localization of peptidoglycan synthesis to be determined by fluorescence microscopy. Herein we describe the chemical synthesis of an OregonGreen488-labelled FDAA (OGDA). We also demonstrate that OGDA can be efficiently incorporated into the PG of Gram-positive and some Gram-negative bacteria, and imaged by super-resolution stimulated emission depletion (STED) nanoscopy at a resolution well below 100 nm.

Keywords: FDAA; FLDA; OGDA; OregonGreen488; peptidoglycan; transpeptidase.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry
  • Amino Acids / metabolism*
  • Fluorescent Dyes / chemistry
  • Fluorescent Dyes / metabolism*
  • Gram-Negative Bacteria / metabolism
  • Microscopy, Fluorescence
  • Molecular Imaging
  • Peptidoglycan / biosynthesis*

Substances

  • Amino Acids
  • Fluorescent Dyes
  • Peptidoglycan