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Assessment of Mitochondrial Protein Glutathionylation as Signaling for CO Pathway

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Mitochondrial Medicine

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1264))

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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 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.

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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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Authors and Affiliations

  1. Chronic Diseases Research Center (CEDOC), Faculdade de Ciências Médicas, Universidade Nova de Lisboa, Campo Mártires da Pátria 130, 1169-056, Lisboa, Portugal

    Ana S. Almeida & Helena L. A. Vieira

  2. Instituto de Biologia Experimental e Tecnológica (IBET), Av. da República, 2780-157, Oeiras, Portugal

    Ana S. Almeida & Helena L. A. Vieira

  3. Instituto de Tecnologia Química e Biológica (ITQB), Universidade Nova de Lisboa, Apt 127, 2781-901, Oeiras, Portugal

    Ana S. Almeida

Authors
  1. Ana S. Almeida
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  2. Helena L. A. Vieira
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Corresponding author

Correspondence to Helena L. A. Vieira .

Editor information

Editors and Affiliations

  1. Midwestern University, Glendale, Arizona, USA

    Volkmar Weissig

  2. ISANH, Paris, France

    Marvin Edeas

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Almeida, A.S., Vieira, H.L.A. (2015). Assessment of Mitochondrial Protein Glutathionylation as Signaling for CO Pathway. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine. Methods in Molecular Biology, vol 1264. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2257-4_30

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  • DOI: https://doi.org/10.1007/978-1-4939-2257-4_30

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2256-7

  • Online ISBN: 978-1-4939-2257-4

  • eBook Packages: Springer Protocols

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