Identification of AHL- and BDSF-Controlled Proteins in Burkholderia cenocepacia by Proteomics

Methods Mol Biol. 2018:1673:193-202. doi: 10.1007/978-1-4939-7309-5_15.

Abstract

We used comparative proteome analysis to determine the target genes of the two quorum sensing (QS) circuits in the opportunistic pathogen Burkholderia cenocepacia: the N-acyl homoserine lactone (AHL)-based CepIR system and the BDSF (B urkholderia diffusible signal factor, cis-2-dodecenoic acid)-based RpfFR system. In this book chapter, we focus on the description of the practical procedure we currently use in the laboratory to perform a sensitive GeLC-MS/MS shotgun proteomics experiment; we also briefly describe the downstream bioinformatic data analysis.

Keywords: Cell fractionation; In-gel tryptic digestion; Mass spectrometry; Quorum sensing; SDS-PAGE.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyl-Butyrolactones / metabolism*
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism*
  • Burkholderia cenocepacia / metabolism*
  • Cell Membrane / metabolism
  • Electrophoresis, Polyacrylamide Gel
  • Fatty Acids, Monounsaturated / metabolism*
  • Intracellular Space / metabolism
  • Mass Spectrometry
  • Peptides / isolation & purification
  • Proteomics / methods*
  • Staining and Labeling
  • Statistics as Topic

Substances

  • 2-dodecenoic acid
  • Acyl-Butyrolactones
  • Bacterial Proteins
  • Fatty Acids, Monounsaturated
  • Peptides