title: Reversible oxidation of glyceraldehyde 3-phosphate dehydrogenase thiols in human lung carcinoma cells by hydrogen peroxide.
abstract: Human lung carcinoma cells (A549) were oxidatively stressed with mildly-toxic or non-toxic amounts of hydrogen peroxide (H2O2, 0.1 mM to 120 mM) for 5 min. Hydrogen peroxide exposure resulted in a dose dependent inhibition of binding (pH 7) of the thiol reagent iodoacetic acid (IAA) to a 38 kDa cell protein. Incubation of cells in saline for 60 min following H2O2 removal restored the ability of IAA to bind to the protein. Treatment with 20 mM dithiothreitol or 2 M urea also restored IAA binding, but 10% Triton X102 or 1 mM Brij 58 had no effect. Increasing to pH 11 during the IAA binding also increased thiol availability. Glyceraldehyde 3-phosphate dehydrogenase (EC 1.2.1.12) has been identified as the protein undergoing thiol/disulfide redox status and enzymic activity changes.
authors: Department of Biochemistry and Biophysics, Oregon State University, Corvallis 97331.
authors: A E
authors: AE
authors: Brodie
authors: D J
authors: DJ
authors: Reed
mesh_terms: D002374:Catalase / Q000494:pharmacology; D002460:Cell Line; D004229:Dithiothreitol / Q000494:pharmacology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D007461:Iodoacetates / Q000378:metabolism; D019807:Iodoacetic Acid; D007700:Kinetics; D008175:Lung Neoplasms / Q000201:enzymology*; D010084:Oxidation-Reduction; D011485:Protein Binding; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D007461:Iodoacetates; D013438:Sulfhydryl Compounds; D006861:Hydrogen Peroxide; D002374:Catalase; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D004229:Dithiothreitol; D019807:Iodoacetic Acid