Establishment of a system for monitoring endoplasmic reticulum redox state in mammalian cells

• Published in: Laboratory investigation Vol. 93; no. 11; pp. 1254 - 1258
• Main Authors: Kanekura, Kohsuke, Ishigaki, Shinsuke, Merksamer, Philip I, Papa, Feroz R, Urano, Fumihiko
• Format: Journal Article
• Language: English
• Published: New York Nature Publishing Group US 01.11.2013; Nature Publishing Group
• Subjects: 631/443/319/1557; 631/80/470/1463; 692/699; Animals; Cell Line; Computer Systems; Diabetes Mellitus - metabolism; Endoplasmic Reticulum - metabolism; Endoplasmic Reticulum Stress; Green Fluorescent Proteins - metabolism; HEK293 Cells; Homeostasis; Humans; Laboratory Medicine; Medicine; Medicine & Public Health; Mice; Nerve Degeneration - metabolism; Oxidation-Reduction; Pathology; Protein Folding; Rats; Recombinant Proteins - metabolism; research-article; Wolfram Syndrome - metabolism; beta cell; redox regulation; Wolfram syndrome; ER stress; neurodegeneration; neuron; diabetes
• ISSN: 0023-6837; 1530-0307
• DOI: 10.1038/labinvest.2013.112

The endoplasmic reticulum (ER) performs a critical role in the oxidative folding of nascent proteins, such that perturbations to ER homeostasis may lead to protein misfolding and subsequent pathological processes. Among the mechanisms for maintaining ER homeostasis is a redox regulation, which is a critical determinant of the fate of ER-stressed cells. Here, we report the establishment of a system for monitoring the ER redox state in mammalian cells. The new ER redox-sensing system was developed based on the previously described monitoring system in yeast. Our system could successfully monitor the dynamic ER redox state in mammalian cells. Using this system, we find that manipulation of ER oxidases changes the ER redox state. The mammalian ER redox-sensing system could be used to study the mechanisms of ER redox regulation and provide a foundation for an approach to develop novel therapeutic modalities for human diseases related to dysregulated ER homeostasis including diabetes, neurodegeneration, and Wolfram syndrome.