Profiling protein expression in Klebsiella pneumoniae with a carbohydrate-based covalent probe

Bioorg Med Chem. 2021 Jan 15:30:115900. doi: 10.1016/j.bmc.2020.115900. Epub 2020 Nov 30.

Abstract

We report the application of a covalent probe based on a d-glucosamine scaffold for the profiling of the bacterial pathogen Klebsiella pneumoniae. Incubation of K. pneumoniae lysates with the probe followed by electrophoretic separation and in-gel fluorescence detection allowed the generation of strain-specific signatures and the differentiation of a carbapenem-resistant strain. The labelling profile of the probe was independent of its anomeric configuration and included several low-abundance proteins not readily detectable by conventional protein staining. Initial target identification experiments by mass spectrometry suggest that target proteins include several carbohydrate-recognising proteins, which indicates that the sugar scaffold may have a role for target recognition.

Keywords: Antimicrobial resistance; Bacterial profiling; Carbohydrate; Covalent probe; Klebsiella.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics*
  • Dose-Response Relationship, Drug
  • Fluorescent Dyes / chemical synthesis
  • Fluorescent Dyes / chemistry*
  • Gene Expression Profiling
  • Glucosamine / chemical synthesis
  • Glucosamine / chemistry*
  • Klebsiella pneumoniae / genetics*
  • Klebsiella pneumoniae / isolation & purification
  • Molecular Structure
  • Structure-Activity Relationship

Substances

  • Bacterial Proteins
  • Fluorescent Dyes
  • Glucosamine