Development of roGFP2-derived redox probes for measurement of the glutathione redox potential in the cytosol of severely glutathione-deficient rml1 seedlings

@article{Aller2013DevelopmentOR,
  title={Development of roGFP2-derived redox probes for measurement of the glutathione redox potential in the cytosol of severely glutathione-deficient rml1 seedlings},
  author={Isabel Aller and Nicolas Rouhier and Andreas J. Meyer},
  journal={Frontiers in Plant Science},
  year={2013},
  volume={4}
}
Glutathione is important for detoxification, as a cofactor in biochemical reactions and as a thiol-redox buffer. [...] Key Method To allow rapid and specific equilibration with the glutathione pool, fusion constructs with human glutaredoxin 1 (GRX1) were generated and characterized in vitro. GRX1-roGFP2-iL proved to be suitable for in vivo redox potential measurements and extends the range of E GSH values that can be measured in vivo with roGFP2-based probes from about -320 mV for GRX1-roGFP2 down to about -210…Expand
Utilizing redox-sensitive GFP fusions to detect in vivo redox changes in a genetically engineered prokaryote
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The previously characterized reduction-oxidation sensitive green fluorescent protein (roGFP2) was used to elucidate the redox changes of the genetically engineered Escherichia coli strain, SHuffle, demonstrating to be under constitutive oxidative stress and responding transcriptionally in an OxyR-dependent manner. Expand
The Arabidopsis glutathione transferases, AtGSTF8 and AtGSTU19 are involved in the maintenance of root redox homeostasis affecting meristem size and salt stress sensitivity.
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The results indicate that atGSTF8 and especially the AtGSTU19 proteins function in the root fine-tuning the redox homeostasis both under control and salt stress conditions. Expand
Exogenously applied salicylic acid maintains redox homeostasis in salt-stressed Arabidopsis gr1 mutants expressing cytosolic roGFP1
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Arabidopsis glutathione reductase 2 is indispensable in plastids, while mitochondrial glutathione is safeguarded by additional reduction and transport systems.
TLDR
The data demonstrate that GR2 is dual-targeted to plastids and mitochondria, but embryo lethality of gr2 null mutants is caused specifically in plastIDS, and a functional triad of GR2, ATM3 and the thioredoxin system in the mitochondria provides resilience to excessive glutathione oxidation. Expand
Arabidopsis glutathione reductase 2 is indispensable in plastids, while mitochondrial glutathione is safeguarded by additional reduction and transport systems
TLDR
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TLDR
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Glutathione--linking cell proliferation to oxidative stress.
TLDR
It is proposed that regulated ROS production linked to glutathione-mediated signalling events are the hallmark of viable cells within a changing and challenging environment. Expand
Quantitative Monitoring of Subcellular Redox Dynamics in Living Mammalian Cells Using RoGFP2-Based Probes.
TLDR
Step-by-step protocols and tips for the successful use of roGFP2, a redox-sensitive variant of the enhanced green fluorescent protein, to monitor changes in glutathione redox balance and hydrogen peroxide homeostasis in the cytosol, peroxisomes, and mitochondria of mammalian cells are provided. Expand
Nuclear thiol redox systems in plants.
TLDR
An overview of the different thiol-based redox regulated proteins in the nucleus that are involved in various pathways including transcriptional regulation, metabolism and signaling is made. Expand
Crosstalk between the redox signalling and the detoxification: GSTs under redox control?
TLDR
High-throughput quantitative real-time PCR revealed that besides overall higher expression levels, SlGSTs were activated more efficiently in 'Moneymaker' due to stresses, resulting in generally higher GST and glutathione peroxidase activities compared to 'Ailsa Craig'. Expand
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References

SHOWING 1-10 OF 68 REFERENCES
Redox-sensitive GFP in Arabidopsis thaliana is a quantitative biosensor for the redox potential of the cellular glutathione redox buffer.
TLDR
The results with roGFP2 as an artificial GRX target further suggest that redox-triggered changes of biologic processes might be linked directly to the glutathione redox potential via GRX as the mediator. Expand
The NADPH-dependent thioredoxin system constitutes a functional backup for cytosolic glutathione reductase in Arabidopsis
TLDR
Interestingly, dynamic reduction of GSSG formed during induced oxidative stress in gr1 mutants is still possible, although significantly delayed compared with wild-type plants, and it is inferred that there is functional redundancy in this critical pathway. Expand
Real-time imaging of the intracellular glutathione redox potential
TLDR
It is demonstrated that the fusion of human glutaredoxin-1 to roGFP2 facilitates specific real-time equilibration between the sensor protein and the glutathione redox couple, which facilitated the observation of redox changes associated with growth factor availability, cell density, mitochondrial depolarization, respiratory burst activity and immune receptor stimulation. Expand
Redesign of Genetically Encoded Biosensors for Monitoring Mitochondrial Redox Status in a Broad Range of Model Eukaryotes
TLDR
The redesigned probe thus allows extending in vivo observations of mitochondrial redox homeostasis to important nonmammalian model organisms, particularly plants and insects. Expand
Confocal imaging of glutathione redox potential in living plant cells
TLDR
Specific interaction of both roGFPs with glutaredoxin in vitro strongly suggests that in situ both variants preferentially act as sensors for the glutathione redox potential, suggesting that roG FP2 is a reliable probe for dynamic redox imaging in planta. Expand
Glutathione homeostasis and redox-regulation by sulfhydryl groups
TLDR
The central role of glutathione is emphasized by its involvement in signaling processes and the crosstalk of redox signaling processes with other means of signaling including protein glutathionylation and control of transcription factors. Expand
Investigating Mitochondrial Redox Potential with Redox-sensitive Green Fluorescent Protein Indicators*
TLDR
By substitution of surface-exposed residues on the Aequorea victoria green fluorescent protein with cysteines in appropriate positions to form disulfide bonds, reduction-oxidation-sensitive GFPs (roGFPs) have been created and targeted to the mitochondria in HeLa cells. Expand
Development of a family of redox-sensitive green fluorescent protein indicators for use in relatively oxidizing subcellular environments.
TLDR
A new family of GFP-based redox indicators (roGFP1-iX) in which the thermodynamic stability of the disulfide is substantially lowered by insertion of a single amino acid into the main chain, adjacent to cysteine 147, which result in indicators with midpoint potentials of -229 to -246 mV and are better suited for study of relatively oxidizing subcellular compartments. Expand
Imaging Dynamic Redox Changes in Mammalian Cells with Green Fluorescent Protein Indicators*
TLDR
The response of the redox probes under physiological redox changes during superoxide bursts in macrophage cells, hyperoxic and hypoxic conditions, and in responses to H2O2-stimulating agents, e.g. epidermal growth factor and lysophosphatidic acid is investigated. Expand
A fluorometer-based method for monitoring oxidation of redox-sensitive GFP (roGFP) during development and extended dark stress.
TLDR
F fluorometer measurements of roGFP can be used for extended measurements enabling the processing of multiple samples and may be applicable to the analysis of redox changes in response to other stresses or to various mutants. Expand
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