State-selective metabolic labeling of cellular proteins

ACS Chem Biol. 2012 Aug 17;7(8):1326-30. doi: 10.1021/cb300238w. Epub 2012 Jun 12.

Abstract

Transcriptional activity from a specified promoter can provide a useful marker for the physiological state of a cell. Here we introduce a method for selective tagging of proteins made in cells in which specified promoters are active. Tagged proteins can be modified with affinity reagents for enrichment or with fluorescent dyes for visualization. The method allows state-selective analysis of the proteome, whereby proteins synthesized in predetermined physiological states can be identified. The approach is demonstrated by proteome-wide labeling of bacterial proteins upon activation of the P(BAD) promoter and the SoxRS regulon and provides a basis for analysis of more complex systems including spatially heterogeneous microbial cultures and biofilms.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arabinose / genetics
  • Bacterial Proteins / chemistry
  • Biofilms
  • Cell Survival
  • Cloning, Molecular
  • Escherichia coli / metabolism
  • Fluorescent Dyes / chemistry
  • Gene Expression Regulation
  • Genetic Techniques
  • Ligands
  • Magnetic Resonance Spectroscopy / methods
  • Models, Biological
  • Models, Genetic
  • Promoter Regions, Genetic
  • Proteomics / methods
  • Regulon
  • Transcription, Genetic*

Substances

  • Bacterial Proteins
  • Fluorescent Dyes
  • Ligands
  • Arabinose