title: Different consequences of reactions with hydrogen peroxide and t-butyl hydroperoxide in the hyperoxidative inactivation of rat peroxiredoxin-4.
abstract: Eukaryotic typical 2-Cys type peroxiredoxin (Prx) is inactivated by hyperoxidation of the peroxidatic cysteine to a sulphinic acid in a catalytic cycle-dependent manner. This inactivation process has been well documented for cytosolic isoforms of Prx. However, such a hyperoxidative inactivation has not fully been investigated in Prx-4, a secretable endoplasmic reticulum-resident isoform, in spite of being a typical 2-Cys type, and details of this process are reported herein. As has been observed in many peroxiredoxins, the peroxidase activity of Prx-4 was almost completely inhibited in the reaction with t-butyl hydroperoxide. On the other hand, when H(2)O(2) was used as the substrate, the peroxidase activity significantly remained after oxidative damage. In spite of these different consequences, mass spectrometric analyses indicated that both reactions resulted in the same oxidative damage, i.e. sulphinic acid formation at the peroxidatic cysteine, suggesting that another cysteine in the active site confers the peroxidase activity. As suggested by the analyses using cysteine-substituted mutants sulphinic acid formation at the peroxidatic cysteine may play a role in the development of the possible alternative mechanism, thereby sustaining the peroxidase activity that prefers H(2)O(2).
authors.affiliation: Division of Molecular Cell Biology, Department of Biomolecular Sciences, Faculty of Medicine, Saga University, 5-1-1 Nabeshima, Saga 849-8501, Japan. yikeda@med.saga-u.ac.jp
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Ikeda
authors.forename: Miyako
authors.initials: M
authors.lastname: Nakano
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Ihara
authors.forename: Ritsu
authors.initials: R
authors.lastname: Ito
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Taniguchi
authors.forename: Junichi
authors.initials: J
authors.lastname: Fujii
mesh_terms: D000818:Animals; D006861:Hydrogen Peroxide / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D054464:Peroxiredoxins / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020122:tert-Butylhydroperoxide / Q000737:chemistry* / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D020122:tert-Butylhydroperoxide; D006861:Hydrogen Peroxide; D054464:Peroxiredoxins