A PDI-catalyzed thiol-disulfide switch regulates the production of hydrogen peroxide by human Ero1

Thomas Ramming; Masaki Okumura; Shingo Kanemura; Sefer Baday; Julia Birk; Suzette Moes; Martin Spiess; Paul Jenö; Simon Bernèche; Kenji Inaba; Christian Appenzeller-Herzog

doi:10.1016/j.freeradbiomed.2015.02.011

A PDI-catalyzed thiol-disulfide switch regulates the production of hydrogen peroxide by human Ero1

Thomas Ramming
, Masaki Okumura
, Shingo Kanemura
, Sefer Baday
, Julia Birk
, Suzette Moes
, Martin Spiess
, Paul Jenö
, Simon Bernèche
, Kenji Inaba
, Christian Appenzeller-Herzog^*

^*Bu çalışma için yazışmadan sorumlu yazar

University of Basel
Tohoku University
Berufsfachschule Gesundheit Baselland

Araştırma sonucu: Dergiye katkı › Makale › bilirkişi

67 Atıf (Scopus)

Özet

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 Cys²⁰⁸-Cys²⁴¹ 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 Cys²⁰⁸/Cys²⁴¹-dependent mixed-disulfide complex with Ero1α. The H₂O₂-detoxifying glutathione peroxidase 8 also binds to the Cys²⁰⁸/Cys²⁴¹ 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.

Orijinal dil	İngilizce
Sayfa (başlangıç-bitiş)	361-372
Sayfa sayısı	12
Dergi	Free Radical Biology and Medicine
Hacim	83
DOI'lar	https://doi.org/10.1016/j.freeradbiomed.2015.02.011
Yayın durumu	Yayınlandı - 19 May 2015
Harici olarak yayınlandı	Evet

Bibliyografik not

Publisher Copyright:
© 2015 Elsevier Inc. All rights reserved.

Finansman

This paper is dedicated to Emil, who was born during the finalization of the manuscript. We are grateful to Alex Odermatt for generous support. We also thank Adam Lister for editing of the manuscript; Lloyd Ruddock, Neil Bulleid, Jan Riemer, Miklos Geiszt, Roberto Sitia, Hans-Peter Hauri, and Ari Helenius for sharing reagents; and PRACE for awarding us access to resource Lindgren based in Sweden at the PDC Center for High Performance Computing. This work was supported by a Ph.D. fellowship from the Boehringer Ingelheim Fonds (to T.R.), a Grant-in-Aid for JSPS Fellows (to M.O.), a grant for Next Generation World-Leading Researchers from MEXT (to K.I.), and the Swiss National Science Foundation (Ambizione), the University of Basel, and the Freiwillige Akademische Gesellschaft (all to C.A.H.). The synchrotron radiation experiment was performed at BL45XU in SPring-8 with the approval of RIKEN (Proposal No. 2014A1345).

Finansörler	Finansör numarası
Universität Basel
Freiwillige Akademische Gesellschaft
Boehringer Ingelheim Fonds
Japan Society for the Promotion of Science	15H04335, 15J06349, 13J04030
Ministry of Education, Culture, Sports, Science and Technology
Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung	144111, 142964

Belgeye Erişim

10.1016/j.freeradbiomed.2015.02.011

Diğer Dosyalar ve Linkler

Link to publication in Scopus

Alıntı Yap

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title = "A PDI-catalyzed thiol-disulfide switch regulates the production of hydrogen peroxide by human Ero1",

abstract = "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 Cys208-Cys241 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 Cys208/Cys241-dependent mixed-disulfide complex with Ero1α. The H2O2-detoxifying glutathione peroxidase 8 also binds to the Cys208/Cys241 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.",

keywords = "Disulfide bond formation, Endoplasmic reticulum, Ero1, Hydrogen peroxide, Oxidative folding, Peroxidase: Free radicals",

author = "Thomas Ramming and Masaki Okumura and Shingo Kanemura and Sefer Baday and Julia Birk and Suzette Moes and Martin Spiess and Paul Jen{\"o} and Simon Bern{\`e}che and Kenji Inaba and Christian Appenzeller-Herzog",

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month = may,

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doi = "10.1016/j.freeradbiomed.2015.02.011",

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pages = "361--372",

journal = "Free Radical Biology and Medicine",

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T1 - A PDI-catalyzed thiol-disulfide switch regulates the production of hydrogen peroxide by human Ero1

AU - Ramming, Thomas

AU - Okumura, Masaki

AU - Kanemura, Shingo

AU - Baday, Sefer

AU - Birk, Julia

AU - Moes, Suzette

AU - Spiess, Martin

AU - Jenö, Paul

AU - Bernèche, Simon

AU - Inaba, Kenji

AU - Appenzeller-Herzog, Christian

PY - 2015/5/19

Y1 - 2015/5/19

N2 - 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 Cys208-Cys241 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 Cys208/Cys241-dependent mixed-disulfide complex with Ero1α. The H2O2-detoxifying glutathione peroxidase 8 also binds to the Cys208/Cys241 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.

AB - 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 Cys208-Cys241 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 Cys208/Cys241-dependent mixed-disulfide complex with Ero1α. The H2O2-detoxifying glutathione peroxidase 8 also binds to the Cys208/Cys241 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.

KW - Disulfide bond formation

KW - Endoplasmic reticulum

KW - Ero1

KW - Hydrogen peroxide

KW - Oxidative folding

KW - Peroxidase: Free radicals

UR - https://www.scopus.com/pages/publications/84929325257

U2 - 10.1016/j.freeradbiomed.2015.02.011

DO - 10.1016/j.freeradbiomed.2015.02.011

M3 - Article

C2 - 25697776

AN - SCOPUS:84929325257

SN - 0891-5849

VL - 83

SP - 361

EP - 372

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

ER -

A PDI-catalyzed thiol-disulfide switch regulates the production of hydrogen peroxide by human Ero1

Özet

Bibliyografik not

Finansman

Belgeye Erişim

Diğer Dosyalar ve Linkler

Parmak izi

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