Measure of Peptidoglycan Hydrolase Activity

Methods Mol Biol. 2017;1615:151-158. doi: 10.1007/978-1-4939-7033-9_12.

Abstract

Most gene clusters encoding multiprotein complexes of the bacterial cell envelope, such as conjugation and secretion systems, Type IV pili, and flagella, bear a gene encoding an enzyme with peptidoglycan hydrolase activity. These enzymes are usually glycoside hydrolases that cleave the glycan chains of the peptidoglycan. Their activities are spatially controlled to avoid cell lysis and to create localized rearrangement of the cell wall. This is assured by interaction with the structural subunits of the apparatus. Here we describe protocols to test the peptidoglycan hydrolase activity of these proteins in vitro and in solution.

Keywords: Cell wall; Localized degradation; Lytic transglycosylase; Peptidoglycan; Remazol blue.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Enzyme Activation
  • Hydrolysis
  • N-Acetylmuramoyl-L-alanine Amidase / chemistry*
  • N-Acetylmuramoyl-L-alanine Amidase / metabolism
  • Nephelometry and Turbidimetry*
  • Peptidoglycan / chemistry*
  • Peptidoglycan / isolation & purification
  • Peptidoglycan / metabolism
  • Staining and Labeling

Substances

  • Peptidoglycan
  • N-Acetylmuramoyl-L-alanine Amidase