Assessing the effect of reactive oxygen species on Escherichia coli using a metabolome approach.

A two-dimensional thin-layer chromatographic analysis of [14C]-labelled metabolites in Escherichia coli was employed to follow metabolic shifts in response to superoxide stress. Steady-state challenge with paraquat at concentrations inducing SoxRS-controlled genes resulted in several alterations in metabolite pools, including a striking increase in valine concentration. Elevated valine levels, together with increased glutathione and alkylperoxidase, are proposed to constitute an intracellular protection mechanism against reactive oxygen species. As shown by this example of metabolome analysis, novel cellular responses to environmental challenge can be revealed by following the total complement of metabolites in a cell.

[1]  F. Blattner,et al.  Global regulation of gene expression in Escherichia coli , 1993, Journal of bacteriology.

[2]  M. Elskens,et al.  Metabolism and functions of glutathione in micro-organisms. , 1993, Advances in microbial physiology.

[3]  J. Gebicki,et al.  Formation of peroxides in amino acids and proteins exposed to oxygen free radicals. , 1993, The Biochemical journal.

[4]  T. Ferenci,et al.  Effect of Slow Growth on Metabolism of Escherichia coli, as Revealed by Global Metabolite Pool (“Metabolome”) Analysis , 1998, Journal of bacteriology.

[5]  I. Fridovich,et al.  Fumarase C, the stable fumarase of Escherichia coli, is controlled by the soxRS regulon. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[6]  J. Eaton,et al.  Bacterial glutathione: a sacrificial defense against chlorine compounds , 1996, Journal of bacteriology.

[7]  S. Farr,et al.  Oxidative stress responses in Escherichia coli and Salmonella typhimurium. , 1991, Microbiological reviews.

[8]  T. Ferenci,et al.  Differential expression of mal genes under cAMP and endogenous inducer control in nutrient‐stressed Escherichia coli , 1995, Molecular microbiology.

[9]  B. Ames,et al.  An alkyl hydroperoxide reductase from Salmonella typhimurium involved in the defense of DNA against oxidative damage. Purification and properties. , 1989, The Journal of biological chemistry.

[10]  L. Wodicka,et al.  Genome-wide expression monitoring in Saccharomyces cerevisiae , 1997, Nature Biotechnology.

[11]  R. Hill,et al.  [17] Fumarase: [EC 4.2.1.2 l-Malate hydro-lyase] , 1969 .

[12]  P. Loewen.  Levels of glutathione in Escherichia coli. , 1979, Canadian journal of biochemistry.

[13]  T. Ferenci,et al.  Derepression of LamB protein facilitates outer membrane permeation of carbohydrates into Escherichia coli under conditions of nutrient stress , 1993, Journal of bacteriology.

[14]  O. Brown,et al.  Dihydroxyacid dehydratase: the site of hyperbaric oxygen poisoning in branch-chain amino acid biosynthesis. , 1978, Biochemical and biophysical research communications.

[15]  D. Hochstrasser,et al.  From Proteins to Proteomes: Large Scale Protein Identification by Two-Dimensional Electrophoresis and Arnino Acid Analysis , 1996, Bio/Technology.

[16]  D. Tempest,et al.  Influence of environment on the content and composition of microbial free amino acid pools. , 1970, Journal of general microbiology.

[17]  Jeffrey H. Miller.  Experiments in molecular genetics , 1972 .

[18]  B. Demple,et al.  Redox signaling and gene control in the Escherichia coli soxRS oxidative stress regulon--a review. , 1996, Gene.

[19]  R. Dean,et al.  Biological fate of amino acid, peptide and protein hydroperoxides. , 1995, The Biochemical journal.

[20]  P. Apontoweil,et al.  Glutathione biosynthesis in Escherichia coli K 12. Properties of the enzymes and regulation. , 1975, Biochimica et biophysica acta.

[21]  F. Neidhardt,et al.  Escherichia coli proteome analysis using the gene‐protein database , 1997, Electrophoresis.