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Real-Time Monitoring of Hydrogen Peroxide Levels in Yeast and Mammalian Cells

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Metabolic Reprogramming

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2675))

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Abstract

Hydrogen peroxide (H2O2) is an important signaling molecule involved in regulating antioxidative transcriptional responses, cellular differentiation, and hypoxia response. H2O2 generation and signaling are highly localized processes. Understanding the dynamics of this molecule inside intact cells with subcompartmental resolution is instrumental to unravel its role in cellular signaling. Different genetically encoded fluorescent sensors have been developed over the last few years that enable such non-disruptive monitoring with high spatiotemporal resolution. In this chapter, we describe the use of these genetically encoded sensors to directly monitor H2O2 dynamics in yeast and cultured mammalian cells.

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Acknowledgments

The Deutsche Forschungsgemeinschaft (DFG) funds research in the laboratory of J.R. through grants RI2150/2-2-project number 251546152, RI2150/5-1-project number 435235019, CRC1218 / TP B02-project number 269925409, and RTG2550/1-project number 411422114. G.C. is currently supported by a DFG Walter Benjamin fellowship (CA 2559/1-1) and a MSFHR Research Trainee fellowship (RT-2020-0517). Figure design and original hand-drawn artworks are by G.C.

Author information

Author notes

  1. Gaetano Calabrese and Lianne J. H. C. Jacobs have contributed equally for this chapter.

Authors and Affiliations

  1. Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada

    Gaetano Calabrese

  2. Department for Chemistry, Institute of Biochemistry, University of Cologne, Cologne, Germany

    Lianne J. H. C. Jacobs & Jan Riemer

  3. Cologne Excellence Cluster on Cellular Stress Responses in Aging-Associated Diseases (CECAD), University of Cologne, Cologne, Germany

    Jan Riemer

Authors
  1. Gaetano Calabrese
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  2. Lianne J. H. C. Jacobs
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  3. Jan Riemer
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Corresponding author

Correspondence to Jan Riemer .

Editor information

Editors and Affiliations

  1. Leeds Institute of Medical Research, St. James’s University Hospital, University of Leeds, Leeds, UK

    Salvatore Papa

  2. Center for Genome Engineering and Maintenance, Department of Life Sciences, College of Health, Medicine and Life Sciences, Brunel University London, London, UK

    Concetta Bubici

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Calabrese, G., Jacobs, L.J.H.C., Riemer, J. (2023). Real-Time Monitoring of Hydrogen Peroxide Levels in Yeast and Mammalian Cells. In: Papa, S., Bubici, C. (eds) Metabolic Reprogramming. Methods in Molecular Biology, vol 2675. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3247-5_12

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  • DOI: https://doi.org/10.1007/978-1-0716-3247-5_12

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3246-8

  • Online ISBN: 978-1-0716-3247-5

  • eBook Packages: Springer Protocols

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