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Assessment of Mitochondrial Protein Glutathionylation as Signaling for CO Pathway

  • Ana S. Almeida
  • Helena L. A. VieiraEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1264)

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.

Key words

Glutathionylation Carbon monoxide Mitochondria Glutathione Immunoprecipitation 

Notes

Acknowledgments

This work was supported by the Portuguese Fundação para a Ciência e a Tecnologia (FCT-ANR/NEU-NMC/0022/2012) and the Portuguese Fundação para a Ciência e a Tecnologia for ASA’s SFRH/BD/78440/2011 fellowship.

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Chronic Diseases Research Center (CEDOC), Faculdade de Ciências MédicasUniversidade Nova de LisboaLisboaPortugal
  2. 2.Instituto de Biologia Experimental e Tecnológica (IBET)OeirasPortugal
  3. 3.Instituto de Tecnologia Química e Biológica (ITQB)Universidade Nova de LisboaOeirasPortugal

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