Abstract
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 specific antibody coupled to a solid matrix that binds to glutathione antigen. The precipitated protein is further identified and quantified by immunoblotting analysis.
Original language | English |
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Title of host publication | Probing Mitochondrial Function |
Publisher | Springer New York |
Pages | 343-350 |
Number of pages | 8 |
Volume | 1 |
ISBN (Electronic) | 9781493922574 |
ISBN (Print) | 9781493922567 |
DOIs | |
Publication status | Published - 28 Jan 2015 |
Keywords
- Carbon monoxide
- Glutathione
- Glutathionylation
- Immunoprecipitation
- Mitochondria