Protein levels of Escherichia coli thioredoxins and glutaredoxins and their relation to null mutants, growth phase, and function.
Levels of Escherichia coli thioredoxin 1 (Trx1), Trx2, glutaredoxin 1 (Grx1), Grx2, and Grx3 have been determined by novel sensitive sandwich enzyme-linked immunosorbent assay. In a wild type strain, levels of Trx1 increased from the exponential to the stationary phase of growth (1.5-fold to 3400 ng/mg), as did levels of Grx2 (from approximately 2500 to approximately 8000 ng/mg). Grx3 and Trx2 levels were quite stable during growth ( approximately 4500 and approximately 200 ng/mg, respectively). Grx1 levels decreased from approximately 600 ng/mg at the exponential phase to approximately 285 ng/mg at the stationary phase. A large elevation of Grx1 (20-30-fold), was observed in null mutants for the thioredoxin system whereas levels of the other redoxins in all combinations of examined null mutants barely exceeded a 2-3-fold increase. Measurements of thymidine incorporation in newly synthesized DNA suggested that mainly Grx1 and, to a lesser extent, Trx1 contribute to the reduction of ribonucleotides. All glutaredoxin species were elevated in catalase-deficient strains, implying an antioxidant role for the glutaredoxins. Trx1, Trx2, and Grx1 levels increased after exposure to hydrogen peroxide and decreased after exposure to mercaptoethanol. The levels of Grx2 and Grx3 behaved exactly the opposite, suggesting that the transcription factor OxyR does not regulate their expression.
Potamitou
Aristi
A
Medical Nobel Institute for Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden.
Holmgren
Arne
A
Vlamis-Gardikas
Alexios
A
D001149:Arsenates / Q000494:pharmacology; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D054477:Glutaredoxins; D005980:Glutathione Reductase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D008623:Mercaptoethanol / Q000494:pharmacology; D009154:Mutation; D010088:Oxidoreductases; D011506:Proteins / Q000378:metabolism*; D013879:Thioredoxins / Q000378:metabolism*; D013936:Thymidine / Q000378:metabolism
D001149:Arsenates; D054477:Glutaredoxins; D011506:Proteins; D013879:Thioredoxins; D008623:Mercaptoethanol; D006861:Hydrogen Peroxide; D010088:Oxidoreductases; D002374:Catalase; D005980:Glutathione Reductase; C025657:arsenic acid; D013936:Thymidine