Journal article
A PDI-catalyzed thiol-disulfide switch regulates the production of hydrogen peroxide by human Ero1.
- Ramming T Division of Molecular & Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, 4056 Basel, Switzerland.
- Okumura M Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai 980-8577, Japan.
- Kanemura S Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai 980-8577, Japan.
- Baday S Swiss Institutes of Bioinformatics, University of Basel, 4056 Basel, Switzerland; Biozentrum, University of Basel, 4056 Basel, Switzerland.
- Birk J Division of Molecular & Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, 4056 Basel, Switzerland.
- Moes S Biozentrum, University of Basel, 4056 Basel, Switzerland.
- Spiess M Biozentrum, University of Basel, 4056 Basel, Switzerland.
- Jenö P Biozentrum, University of Basel, 4056 Basel, Switzerland.
- Bernèche S Swiss Institutes of Bioinformatics, University of Basel, 4056 Basel, Switzerland; Biozentrum, University of Basel, 4056 Basel, Switzerland.
- Inaba K Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, Sendai 980-8577, Japan.
- Appenzeller-Herzog C Division of Molecular & Systems Toxicology, Department of Pharmaceutical Sciences, University of Basel, 4056 Basel, Switzerland. Electronic address: Christian.Appenzeller@sbl.ch.
- 2015-02-21
Published in:
- Free radical biology & medicine. - 2015
Disulfide bond formation
Endoplasmic reticulum
Ero1
Hydrogen peroxide
Oxidative folding
Peroxidase: Free radicals
Apoptosis
Blotting, Western
Catalysis
Catalytic Domain
Cell Proliferation
Cells, Cultured
Disulfides
Endoplasmic Reticulum
Flavin-Adenine Dinucleotide
Fluorescent Antibody Technique
HeLa Cells
Humans
Hydrogen Peroxide
Membrane Glycoproteins
Membrane Proteins
Molecular Chaperones
Oxidation-Reduction
Oxidoreductases
Oxygen
Protein Conformation
Protein Disulfide-Isomerases
RNA, Messenger
RNA, Small Interfering
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
English
Oxidative folding in the endoplasmic reticulum (ER) involves ER oxidoreductin 1 (Ero1)-mediated disulfide formation in protein disulfide isomerase (PDI). In this process, Ero1 consumes oxygen (O2) and releases hydrogen peroxide (H2O2), but none of the published Ero1 crystal structures reveal any potential pathway for entry and exit of these reactants. We report that additional mutation of the Cys(208)-Cys(241) disulfide in hyperactive Ero1α (Ero1α-C104A/C131A) potentiates H2O2 production, ER oxidation, and cell toxicity. This disulfide clamps two helices that seal the flavin cofactor where O2 is reduced to H2O2. Through its carboxyterminal active site, PDI unlocks this seal by forming a Cys(208)/Cys(241)-dependent mixed-disulfide complex with Ero1α. The H2O2-detoxifying glutathione peroxidase 8 also binds to the Cys(208)/Cys(241) loop region. Supported by O2 diffusion simulations, these data describe the first enzymatically controlled O2 access into a flavoprotein active site, provide molecular-level understanding of Ero1α regulation and H2O2 production/detoxification, and establish the deleterious consequences of constitutive Ero1 activity.
- Language
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- English
- Open access status
- closed
- Identifiers
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- DOI 10.1016/j.freeradbiomed.2015.02.011
- PMID 25697776
- Persistent URL
- https://sonar.ch/global/documents/232141
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