title: Reexamination of the role of interplay between glutathione and protein disulfide isomerase.
abstract: Protein disulfide isomerase (PDI) has an essential role in the process of disulfide bond formation, where it catalyzes disulfide bond formation, reduction, and isomerization. It is thought that the major route for oxidizing dithiols in folding proteins to disulfides is via Ero1-mediated oxidation of PDI. Since the discovery of Ero1, the role of glutathione in disulfide bond formation has been downplayed. In this study, the role of glutathione in disulfide bond formation was reexamined. Here we have studied in vitro the kinetics of the glutathione-mediated oxidation and reduction of the catalytic a domains of human PDI and yeast Pdi1p. The results obtained from stopped-flow and quenched-flow experiments show that the reactions of PDI and Pdi1p are faster and more complex than previously thought. Our results suggest that the kinetics of oxidation of PDI and Pdi1p by oxidized glutathione are remarkably similar, whereas the kinetics of reduction by reduced glutathione shows clear differences. The data generated here on the rapid reactivity of PDI towards glutathione suggest that reevaluation is required for several aspects of the field of catalyzed disulfide bond formation, including the potential physiological role of glutathione.
authors: Department of Biochemistry, University of Oulu, PO Box 3000, Oulu 90014, Finland.
authors: Anna-Kaisa
authors: AK
authors: Lappi
authors: Lloyd W
authors: LW
authors: Ruddock
mesh_terms: D002384:Catalysis; D020134:Catalytic Domain; D004220:Disulfides / Q000737:chemistry / Q000378:metabolism; D005978:Glutathione / Q000737:chemistry* / Q000378:metabolism*; D019803:Glutathione Disulfide / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008954:Models, Biological; D010084:Oxidation-Reduction; D019704:Protein Disulfide-Isomerases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: D004220:Disulfides; D019704:Protein Disulfide-Isomerases; D005978:Glutathione; D019803:Glutathione Disulfide