Disulphide production by Ero1a-PDI relay is rapid and effectively regulated

TY - JOUR

T1 - Disulphide production by Ero1a-PDI relay is rapid and effectively regulated

AU - Appenzeller-Herzog, Christian

AU - Riemer, Jan

AU - Zito, Ester

AU - Chin, King-Tung

AU - Ron, David

AU - Spiess, Martin

AU - Ellgaard, Lars

PY - 2010/10/6

Y1 - 2010/10/6

N2 - The molecular networks that control endoplasmic reticulum (ER) redox conditions in mammalian cells are incompletely understood. Here, we show that after reductive challenge the ER steady-state disulphide content is restored on a time scale of seconds. Both the oxidase Ero1α and the oxidoreductase protein disulphide isomerase (PDI) strongly contribute to the rapid recovery kinetics, but experiments in ERO1-deficient cells indicate the existence of parallel pathways for disulphide generation. We find PDI to be the main substrate of Ero1α, and mixed-disulphide complexes of Ero1 primarily form with PDI, to a lesser extent with the PDI-family members ERp57 and ERp72, but are not detectable with another homologue TMX3. We also show for the first time that the oxidation level of PDIs and glutathione is precisely regulated. Apparently, this is achieved neither through ER import of thiols nor by transport of disulphides to the Golgi apparatus. Instead, our data suggest that a dynamic equilibrium between Ero1-and glutathione disulphide-mediated oxidation of PDIs constitutes an important element of ER redox homeostasis.

AB - The molecular networks that control endoplasmic reticulum (ER) redox conditions in mammalian cells are incompletely understood. Here, we show that after reductive challenge the ER steady-state disulphide content is restored on a time scale of seconds. Both the oxidase Ero1α and the oxidoreductase protein disulphide isomerase (PDI) strongly contribute to the rapid recovery kinetics, but experiments in ERO1-deficient cells indicate the existence of parallel pathways for disulphide generation. We find PDI to be the main substrate of Ero1α, and mixed-disulphide complexes of Ero1 primarily form with PDI, to a lesser extent with the PDI-family members ERp57 and ERp72, but are not detectable with another homologue TMX3. We also show for the first time that the oxidation level of PDIs and glutathione is precisely regulated. Apparently, this is achieved neither through ER import of thiols nor by transport of disulphides to the Golgi apparatus. Instead, our data suggest that a dynamic equilibrium between Ero1-and glutathione disulphide-mediated oxidation of PDIs constitutes an important element of ER redox homeostasis.

KW - Faculty of Science

KW - disulphide-bond formation, endoplasmic reticulum, Ero1, glutathione, protein disulphide isomerase

U2 - 10.1038/emboj.2010.203

DO - 10.1038/emboj.2010.203

M3 - Journal article

C2 - 20802462

SN - 0261-4189

VL - 29

SP - 3318

EP - 3329

JO - EMBO Journal

JF - EMBO Journal

IS - 19

ER -