title: Regulation of the activity of glyceraldehyde 3-phosphate dehydrogenase by glutathione and H2O2.
abstract: The activity lost during storage of a solution of muscle glyceraldehyde 3-phosphate dehydrogenase was rapidly restored on adding a thiol compound, but not arsenite or azide. On treatment with H2O2, the enzyme was partially inactivated and complete loss of activity occurred in the presence of glutathione. Samples of the enzyme pretreated with glutathione followed by removal of the thiol compound by filtration on a Sephadex column showed both full activity and its complete loss on adding H2O2, in the absence of added glutathione. Most of the activity was restored when the H2O2-inactivated enzyme was incubated with glutathione (25 mM) or dithiothreitol (5 mM) whereas arsenite or azide were partly effective and ascorbate was ineffective. The need for incubation for a long time with a strong reducing agent for restoration of activity suggests that the oxidized group (disulfide or sulfenate) must be in a masked state in the H2O2-inactivated enzyme. Analysis by SDS-PAGE gave evidence for the formation of a small quantity of glutathione-reversible disulfide-form of the enzyme. Circular dichroic spectra indicated a decrease in alpha-helical content in the inactivated form of the enzyme. The evidence suggest that glutathione and H2O2 can regulate the active state of this enzyme.
authors.affiliation: Department of Biochemistry, Indian Institute of Science, Bangalore.
authors.forename: V V
authors.initials: VV
authors.lastname: Vaidyanathan
authors.forename: P S
authors.initials: PS
authors.lastname: Sastry
authors.forename: T
authors.initials: T
authors.lastname: Ramasarma
mesh_terms: D004220:Disulfides / Q000032:analysis; D004789:Enzyme Activation; D005978:Glutathione / Q000494:pharmacology*; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000187:drug effects*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013048:Specimen Handling; D013438:Sulfhydryl Compounds / Q000494:pharmacology
chemical_list: D004220:Disulfides; D013438:Sulfhydryl Compounds; D006861:Hydrogen Peroxide; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D005978:Glutathione