title: Assessment of mitochondrial protein glutathionylation as signaling for CO pathway.
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.
authors: Chronic Diseases Research Center (CEDOC), Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo Mártires da Pátria 130, 1169-056, Lisboa, Portugal.
authors: Ana S
authors: AS
authors: Almeida
authors: Helena L A
authors: HL
authors: Vieira
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002248:Carbon Monoxide / Q000378:metabolism*; D002458:Cell Fractionation / Q000379:methods; D002540:Cerebral Cortex / Q000166:cytology / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism*; D006801:Humans; D047468:Immunoprecipitation; D008928:Mitochondria / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000378:metabolism*; D010084:Oxidation-Reduction; D017382:Reactive Oxygen Species; D015398:Signal Transduction
chemical_list: D024101:Mitochondrial Proteins; D017382:Reactive Oxygen Species; D002248:Carbon Monoxide; D005978:Glutathione