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Multiple glutathione disulfide removal pathways mediate cytosolic redox homeostasis

Nature Chemical Biology volume 9, pages 119–125 (2013)Cite this article

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Abstract

Glutathione is central to cellular redox chemistry. The majority of glutathione redox research has been based on the chemical analysis of whole-cell extracts, which unavoidably destroy subcellular compartment–specific information. Compartment-specific real-time measurements based on genetically encoded fluorescent probes now suggest that the cytosolic glutathione redox potential is about 100 mV more reducing than previously thought. Using these probes in yeast, we show that even during severe oxidative stress, the cytosolic glutathione disulfide (GSSG) concentration is much more tightly regulated than expected and provides a mechanistic explanation for the discrepancy with conventional measurements. GSSG that is not immediately reduced in the cytosol is rapidly transported into the vacuole by the ABC-C transporter Ycf1. The amount of whole-cell GSSG is entirely dependent on Ycf1 and uninformative about the cytosolic glutathione pool. Applying these insights, we identify Trx2 and Grx2 as efficient backup systems to glutathione reductase for cytosolic GSSG reduction.

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Figure 1: Cytosolic glutathione homeostasis is robustly maintained and involves GSSG compartmentalization.
Figure 2: Cytosolic GSSG is rapidly transported to another subcellular compartment.
Figure 3: Whole-cell GSSG changes are completely dependent on Ycf1-mediated vacuolar accumulation.
Figure 4: Ycf1 mediates rapid compartmentalization of GSSG to the vacuole.
Figure 5: Trx2, but not Trx1, contributes to cytosolic GSSG reduction.
Figure 6: Grx2 has an important backup role in cytosolic GSSG reduction.

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Change history

  • 21 December 2012

    In the version of this article initially published online, equation (5) in the Methods section was written incorrectly. This error has been corrected in the PDF and HTML versions of this article.

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Acknowledgements

B.M. was funded by a European Molecular Biology Organization short-term fellowship (313-2009) and a DKFZ visiting scientist fellowship. Support by the Chica and Heinz Schaller Foundation is gratefully acknowledged. We thank B. Schwappach for critical reading of the manuscript.

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Authors and Affiliations

  1. Division of Redox Regulation, German Cancer Research Center (DKFZ), DKFZ–ZMBH Alliance, Heidelberg, Germany

    Bruce Morgan, Daria Ezeriņa, Theresa N E Amoako & Tobias P Dick

  2. Cellular Biochemistry, University of Kaiserslautern, Kaiserslautern, Germany

    Jan Riemer

  3. Zentrum für Molekulare Biologie der Universität Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany

    Matthias Seedorf

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Contributions

B.M., D.E. and T.N.E.A. performed all experiments. J.R. and M.S. provided materials and technical expertise, and contributed to experimental design. T.P.D. and B.M. conceived the project, analyzed the data and wrote the manuscript.

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Correspondence to Tobias P Dick.

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Morgan, B., Ezeriņa, D., Amoako, T. et al. Multiple glutathione disulfide removal pathways mediate cytosolic redox homeostasis. Nat Chem Biol 9, 119–125 (2013). https://doi.org/10.1038/nchembio.1142

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