Label-Free Quantitation of Ribosomal Proteins from Bacillus subtilis for Antibiotic Research

Methods Mol Biol. 2017:1520:291-306. doi: 10.1007/978-1-4939-6634-9_18.

Abstract

Current research is focusing on ribosome heterogeneity as a response to changing environmental conditions and stresses, such as antibiotic stress. Altered stoichiometry and composition of ribosomal proteins as well as association of additional protein factors are mechanisms for shaping the protein expression profile or hibernating ribosomes. Here, we present a method for the isolation of ribosomes to analyze antibiotic-induced changes in the composition of ribosomes in Bacillus subtilis or other bacteria. Ribosomes and associated proteins are isolated by ultracentrifugation and proteins are identified and quantified using label-free mass spectrometry.

Keywords: Mass spectrometry; Proteomics; Ribosome heterogeneity; Stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anti-Bacterial Agents / pharmacology*
  • Bacillus subtilis / drug effects
  • Bacillus subtilis / growth & development
  • Bacillus subtilis / metabolism*
  • Mass Spectrometry
  • Ribosomal Proteins / metabolism*
  • Ribosomes / metabolism
  • Staining and Labeling*
  • Trypsin / metabolism

Substances

  • Anti-Bacterial Agents
  • Ribosomal Proteins
  • Trypsin