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SLC33A1 exports oxidized glutathione to maintain endoplasmic reticulum redox homeostasis

ABSTRACT: The endoplasmic reticulum (ER) requires an oxidative environment to support the efficient maturation of secretory and membrane proteins. This is in part established by glutathione, a redox-active metabolite present in reduced (GSH) and oxidized (GSSG) forms. The ER maintains a higher GSSG:GSH ratio than the cytosol; however, the mechanisms controlling ER redox balance remain poorly understood. To address this, we developed a method for the rapid immunopurification of the ER, enabling comprehensive profiling of its proteome and metabolome. Combining this approach with CRISPR screening, we identified SLC33A1 as the major ER GSSG exporter in mammalian cells. Loss of SLC33A1 leads to GSSG accumulation in the ER and a liposome-based assay demonstrates that SLC33A1 directly transports GSSG. Cryo-EM structures and molecular dynamics simulations reveal how SLC33A1 binds GSSG and identify residues critical for its transport. Finally, an imbalance in GSSG:GSH ratio induces ER stress and dependency on the ER-associated degradation (ERAD) pathway, driven by a shift in protein disulfide isomerases (PDIs) toward their oxidized forms. Altogether, our work establishes SLC33A1-mediated GSSG export as a key mechanism for ER redox homeostasis and protein maturation.

INSTRUMENT(S):

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Cell Culture

SUBMITTER: Henry Sanford

LAB HEAD: Ekaterina V. Vinogradova

PROVIDER: PXD074465 | Pride | 2026-02-19

REPOSITORIES: Pride

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	20241023_HK_2_81_SL1_01.raw	Raw
	20241023_HK_2_81_SL1_02.raw	Raw
	20241023_HK_2_81_SL1_03.raw	Raw
	20241023_HK_2_81_SL1_04.raw	Raw
	20241023_HK_2_81_SL1_05.raw	Raw

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