Assessment of Mitochondrial Protein Glutathionylation as Signaling for CO Pathway

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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 as 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 languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc
Pages249-257
Number of pages9
DOIs
Publication statusPublished - 2021

Publication series

NameMethods in Molecular Biology
Volume2276
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Carbon monoxide
  • Glutathione
  • Glutathionylation
  • Immunoprecipitation
  • Mitochondria

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