Is proteomics a reliable tool to probe the oxidative folding of bacterial membrane proteins?

Vivianne J Goosens, Ruben A T Mars, Michiel Akeroyd, Andre Vente, Annette Dreisbach, Emma L Denham, Thijs R H M Kouwen, Tjeerd van Rij, Maurien Olsthoorn, Jan Maarten van Dijl

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The oxidative folding of proteins involves disulfide bond formation, which is usually catalyzed by thiol-disulfide oxidoreductases (TDORs). In bacteria, this process takes place in the cytoplasmic membrane and other extracytoplasmic compartments. While it is relatively easy to study oxidative folding of water-soluble proteins on a proteome-wide scale, this has remained a major challenge for membrane proteins due to their high hydrophobicity. Here, we have assessed whether proteomic techniques can be applied to probe the oxidative folding of membrane proteins using the Gram-positive bacterium Bacillus subtilis as a model organism. Specifically, we investigated the membrane proteome of a B. subtilis bdbCD mutant strain, which lacks the primary TDOR pair BdbC and BdbD, by gel-free mass spectrometry. In total, 18 membrane-associated proteins showed differing behavior in the bdbCD mutant and the parental strain. These included the ProA protein involved in osmoprotection. Consistent with the absence of ProA, the bdbCD mutant was found to be sensitive to osmotic shock. We hypothesize that membrane proteomics is a potentially effective approach to profile oxidative folding of bacterial membrane proteins.

Original languageEnglish
Pages (from-to)1159-1164
Number of pages6
JournalAntioxidants & Redox Signaling
Volume18
Issue number10
Early online date14 Feb 2013
DOIs
Publication statusPublished - 1 Apr 2013

Keywords

  • Bacillus subtilis/metabolism
  • Bacterial Proteins/chemistry
  • Electrophoresis, Polyacrylamide Gel
  • Membrane Proteins/chemistry
  • Protein Folding
  • Proteomics/methods

Cite this

Goosens, V. J., Mars, R. A. T., Akeroyd, M., Vente, A., Dreisbach, A., Denham, E. L., Kouwen, T. R. H. M., van Rij, T., Olsthoorn, M., & van Dijl, J. M. (2013). Is proteomics a reliable tool to probe the oxidative folding of bacterial membrane proteins? Antioxidants & Redox Signaling, 18(10), 1159-1164. https://doi.org/10.1089/ars.2012.4664