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Assessment of mitochondrial protein glutathionylation as signaling for CO pathway

Methods in Molecular Biology, ISSN: 1064-3745, Vol: 1264, Page: 343-350
2015
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Book Chapter Description

Protein glutathionylation is a posttranslational process that regulates protein function in response to redox cellular changes. Furthermore, carbon monoxide-induced cellular pathways involve reactive oxygen species (ROS) signaling and mitochondrial protein glutathionylation. Herein, it is described a technique to assess mitochondrial glutathionylation due to low concentrations of CO exposure. Mitochondria are isolated from cell culture or tissue, followed by an immunoprecipitation assay, which allows the capture of any glutathionylated mitochondrial protein using a specifi c antibody coupled to a solid matrix that binds to glutathione antigen. The precipitated protein is further identifi ed and quantifi ed by immunoblotting analysis.

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