Assays for the Enzymes Catalyzing the O-Acetylation of Bacterial Cell Wall Polysaccharides

Methods Mol Biol. 2019:1954:115-136. doi: 10.1007/978-1-4939-9154-9_10.

Abstract

The polysaccharides that comprise bacterial cell walls are commonly O-acetylated. This modification confers resistance to hydrolases of innate immune systems and/or controls endogenous autolytic activity. Herein, we present protocols for the compositional analysis of bacterial cell wall O-acetylation, and assays for monitoring O-acetyltransferases and O-acetylesterases. The assays are amenable for the development of high-throughput screens in search of inhibitors of the respective enzymes.

Keywords: Chitooligosaccharides; O-Acetylation; O-acetylesterase; O-acetyltransferase; Peptidoglycan.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Acetylesterase / metabolism*
  • Acetyltransferases / metabolism*
  • Bacteria / chemistry
  • Bacteria / enzymology
  • Bacteria / metabolism*
  • Bacterial Proteins / metabolism*
  • Cell Wall / chemistry
  • Cell Wall / enzymology
  • Cell Wall / metabolism*
  • Chromatography, High Pressure Liquid / methods
  • Enzyme Assays / methods*
  • Mass Spectrometry / methods
  • Peptidoglycan / chemistry
  • Peptidoglycan / metabolism
  • Polysaccharides, Bacterial / chemistry
  • Polysaccharides, Bacterial / metabolism*

Substances

  • Bacterial Proteins
  • Peptidoglycan
  • Polysaccharides, Bacterial
  • Acetyltransferases
  • Acetylesterase

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