Cited by 894Redox biology and oxidative stressSulfur Compounds in BiologyAdvanced Fluorescence Microscopy Techniques

Real-time imaging of the intracellular glutathione redox potential

Gutscher, Marcus, Pauleau, Anne-Laure, Marty, Laurent, Brach, Thorsten, Wabnitz, Guido H, Samstag, Yvonne, Meyer, Andreas J, Dick, Tobias P·2008·Nature Methods

Abstract

Dynamic analysis of redox-based processes in living cells is now restricted by the lack of appropriate redox biosensors. Conventional redox-sensitive GFPs (roGFPs) are limited by undefined specificity and slow response to changes in redox potential. In this study we demonstrate that the fusion of human glutaredoxin-1 (Grx1) to roGFP2 facilitates specific real-time equilibration between the sensor protein and the glutathione redox couple. The Grx1-roGFP2 fusion protein allowed dynamic live imaging of the glutathione redox potential (E(GSH)) in different cellular compartments with high sensitivity and temporal resolution. The biosensor detected nanomolar changes in oxidized glutathione (GSSG) against a backdrop of millimolar reduced glutathione (GSH) on a scale of seconds to minutes. It facilitated the observation of redox changes associated with growth factor availability, cell density, mitochondrial depolarization, respiratory burst activity and immune receptor stimulation.

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Crossref · 10.1038/nmeth.1212

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  • This recombinant sensor apparently equilibrates more quickly with cellular GSSG than redox-sensitive probes that lack the Grx domain in cis with the redox-sensor domain.

    “This recombinant sensor apparently equilibrates more quickly with cellular GSSG than redox-sensitive probes that lack the Grx domain in cis with the redox-sensor domain.”

    From Redox regulation of mitochondrial function (2012) · cites this paper as [163]

  • A similar biodetector, engineered by combining human Grx with a redox-sensitive GFP, was used to image GSSG-redox changes within human cells.

    “A similar biodetector, engineered by combining human Grx with a redox-sensitive GFP, was used to image GSSG-redox changes within human cells (163).”

    From Redox regulation of mitochondrial function (2012) · cites this paper as [163]

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Authors
Gutscher, M., Pauleau, A.-L., Marty, L., Brach, T., Wabnitz, G. H., Samstag, Y., Meyer, A. J., & Dick, T. P.
Affiliations
  • German Cancer Research Center (DE)
  • Heidelberg University (DE)
Corresponding author
Dick, Tobias P
Venue
Nature Methods
Published
2008-05-11
Volume
5
Issue
6
Pages
553-559
ISSN
  • 1548-7091 (ISSN-L)
  • 1548-7105
OpenAlex
W2100811292
Cited by
894
References
37
Citations by year
2026: 38 · 2025: 51 · 2024: 70 · 2023: 43 · 2022: 72 · 2021: 53 · 2020: 58 · 2019: 67 · 2018: 62 · 2017: 67 · 2016: 50 · 2015: 54 · 2014: 48 · 2013: 45 · 2012: 30
FWCI
11.78
Type
journal article
Language
en
Open access
closed
Locations
  • Nature Methods (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Sulfur Compounds in Biology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
Keywords
glutathione redox potential, glutaredoxin 1, roGFP2, ratiometric biosensor, glutathione disulfide, reduced glutathione, mitochondrial depolarization, respiratory burst, cell density, growth factor signaling
MeSH terms
Biotechnology, Disulfides, Glutathione, HeLa Cells, Humans, Mitochondria, Oxidation-Reduction, Oxygen, Recombinant Fusion Proteins, Sensitivity and Specificity, Thioredoxins, Biosensing Techniques, Apoptosis, Green Fluorescent Proteins, Glutaredoxins

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