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Competition between glutathione and protein thiols for disulphide-bond formation

Nature Cell Biology volume 1, pages 130–135 (1999)Cite this article

Abstract

It has long been assumed that the oxidized form of glutathione, the tripeptide glutamate–cysteine–glycine, is a source of oxidizing equivalents needed for the formation of disulphide bonds in proteins within the endoplasmic reticulum (ER), although the in vivo function of glutathione in the ER has never been studied directly. Here we show that the major pathway for oxidation in the yeast ER, defined by the protein Ero1, is responsible for the oxidation of both glutathione and protein thiols. However, mutation and overexpression studies show that glutathione competes with protein thiols for the oxidizing machinery. Thus, contrary to expectation, cellular glutathione contributes net reducing equivalents to the ER; these reducing equivalents can buffer the ER against transient hyperoxidizing conditions.

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Figure 1: The production of oxidized glutathione (GSSG) is coupled to Ero1 activity.
Figure 2: Growth of ero1-1 cells at high temperature is restored by gsh1::LEU2.
Figure 3: CPY oxidation is restored in the mutant by decreased intracellular glutathione amounts.
Figure 4: Induction of the unfolded-protein response (UPR) is correlated to intracellular glutathione levels.
Figure 5: Diamide blocks CPY transport by generating hyperoxidizing conditions in the ER.
Figure 6: CPY transport in cells is more sensitive to hyperoxidizing conditions than in wild-type cells.
Figure 7: Overview of glutathione and protein disulphide-bond formation in the ER.

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Acknowledgements

We thank H. Ploegh for anti-CPY antibody; R. Sauer and A. Frand for helpful discussions and critical reviews of this manuscript; and all members of the Kaiser laboratory for help and suggestions throughout the course of this project. This work was supported by a grant from the National Institutes of General Medical Sciences (GM46941). J. W. C. was supported by an NIH National Research Service Award (GM18715).

Correspondence and requests for materials should be addressed to C.A.K.

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  1. Department of Biology, Room 68-533, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, 02139, Massachusetts, USA

    John W. Cuozzo & Chris A. Kaiser

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Correspondence to Chris A. Kaiser.

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Cuozzo, J., Kaiser, C. Competition between glutathione and protein thiols for disulphide-bond formation. Nat Cell Biol 1, 130–135 (1999). https://doi.org/10.1038/11047

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