Quantitative in vivo measurement of glutathione in Arabidopsis cells.

@article{Meyer2001QuantitativeIV,
  title={Quantitative in vivo measurement of glutathione in Arabidopsis cells.},
  author={Andreas J. Meyer and Mike J. May and Mark D. Fricker},
  journal={The Plant journal : for cell and molecular biology},
  year={2001},
  volume={27 1},
  pages={
          67-78
        }
}
A new, non-destructive assay is described to quantify cytoplasmic glutathione (GSH) levels in vivo in single cells or populations of cells from Arabidopsis suspension cultures. Cytoplasmic GSH was labelled with monochlorobimane (MCB) in situ to give a fluorescent GSH-bimane (GSB) conjugate. At low (10-100 microM) concentrations of MCB, labelling was mediated by a glutathione S-transferase, which confers specificity for GSH. HPLC analysis of MCB-labelled low molecular-weight thiols showed that… Expand
Control of Demand-Driven Biosynthesis of Glutathione in Green Arabidopsis Suspension Culture Cells1
TLDR
In contrast to the other feeding experiments, exclusion of SO4 2− from the medium abolished the second phase completely, suggesting demand-driven GSH biosynthesis is directly coupled to uptake of SO 4 2− and that the linear increase in fluorescence reflects flux through the entire SO42− assimilation pathway. Expand
Cell-specific measurement of cytosolic glutathione in poplar leaves.
TLDR
In the present study in situ labelling with monochlorobimane (MCB) in combination with confocal laser scanning microscopy was used to quantify GSH in each of the main cell types of poplar leaves from fluorescence of the GSB conjugate formed, suggesting that an appropriate GST was not present within the chloroplasts to catalyse the conjugation reaction. Expand
Increased intracellular H₂O₂ availability preferentially drives glutathione accumulation in vacuoles and chloroplasts.
TLDR
High-performance liquid chromatography and enzymatic assays showed that this correlated tightly with the glutathione redox state, pointing to significant in vivo pools of GSSG in cat2 that were unavailable for MCB labelling. Expand
Restricting glutathione biosynthesis to the cytosol is sufficient for normal plant development.
TLDR
The transgenic lines further show that, consistent with the exclusive plastidic localization of GSH1, gamma-EC is exported from the plastids to supply the cytosol with the immediate precursor for GSH biosynthesis, and that there can be efficient re-import of GSF into the plASTids to allow effective control of G SH biosynthesis through feedback inhibition of Gsh1. Expand
Histochemical quantification of glutathione contents in guard cells of Arabidopsis thaliana
TLDR
MCB staining showed that guard cells of ch1-1, a GSH deficient mutant, accumulated significantly lower GSH content than that of wild type guard cells, and GSH monoethyl ester (GSHmee) restored GSH level in the guard cell of ch 1-1 mutant plants, suggesting that GSHcontent in guard cells can be quantified using MCB dye. Expand
The Problem of Glutathione Determination: a Comparative Study on the Measurement of Glutathione from Plant Cells
The level and redox status of glutathione is a good indicator for the rate of oxidative stress and eco-toxicological injury in plant cells and subcellular organelles. Thus the determination of GSHExpand
Subcellular immunocytochemical analysis detects the highest concentrations of glutathione in mitochondria and not in plastids
TLDR
The present results underline the suggestion that subcellular glutathione concentrations are not controlled by a global mechanism but are controlled on an individual basis and it is therefore not possible to conclude from global biochemicalglutathione analysis on the status of the various organellar pools. Expand
Comparative analyses of glutathione system of vacuoles and leucoplasts isolated from the storage parenchyma cells of dormant red beetroots (Beta vulgaris L.).
TLDR
The idea that the glutathione accumulated by central vacuole seems to contribute to the redox processes and to the detoxification, which can take place in this compartment is suggested. Expand
Sub-cellular distribution of glutathione in an Arabidopsis mutant (vtc1) deficient in ascorbate.
TLDR
It is observed, using immunolabelling techniques, that mitochondria showed the highest density of glutathione labelling in both Col-0 and vtc1 plants during all developmental stages and that the lowest density occurred in the chloroplasts, for both lines. Expand
Modification of intracellular levels of glutathione-dependent formaldehyde dehydrogenase alters glutathione homeostasis and root development.
TLDR
Arabidopsis thaliana mutants with modified levels of FALDH show a significant reduction of root length, and this phenotype correlates with an overall decrease of intracellular GSH levels and alteration of spatial distribution of GSH in the root meristem. Expand
...
1
2
3
4
5
...

References

SHOWING 1-10 OF 49 REFERENCES
Measurement of glutathione levels in intact roots of Arabidopsis
TLDR
Cytoplasmic glutathione concentrations in roots of intact Arabidopsis seedlings were found to be 2–3 mm in most cell types, suggesting that additional cell–cell movement and preferential sequestration of GSB can occur during the detoxification process in an intact system. Expand
Oxidative Stimulation of Glutathione Synthesis in Arabidopsis thaliana Suspension Cultures
TLDR
Data suggest that the induction of GSH synthesis by an oxidative stimulus plays a crucial role in determining the susceptibility of cells to oxidative stress and the degree of protection against oxidative injury. Expand
Kinetic analysis of glutathione in anchored cells with monochlorobimane.
TLDR
Laser cytometric analysis of glutathione in anchored cells provides opportunities to monitor individual cellular responses to a variety of experimental manipulations, such as responses to various toxic insults or the protective effects of gap junction-mediated intercellular communication. Expand
Evidence for posttranscriptional activation of gamma-glutamylcysteine synthetase during plant stress responses.
TLDR
The results demonstrate rapid posttranscriptional activation of Arabidopsis gammaECS and the implications for the mechanisms by which GSH concentrations are regulated during plant-stress responses are discussed. Expand
Detoxification of xenobiotics in plant cells by glutathione conjugation and vacuolar compartmentalization: a fluorescent assay using monochlorobimane
TLDR
A cellular pathway for xenobiotic detoxification in plants is demonstrated, starting with conjugation to glutathione in the cytoplasm, followed by the transport of the conjugates into the vacuole. Expand
The use of monochlorobimane to determine hepatic GSH levels and synthesis.
TLDR
The specific reaction of monochlorobimane with GSH is used to analyze hepatic GSH and determine directly the rate of synthesis of GSH in both cell-free conditions and in cell suspensions by monitoring the increase in fluorescent adduct when mBC1 is present in excess in the incubation. Expand
Kinetic analysis of the intracellular conjugation of monochlorobimane by IC-21 murine macrophage glutathione-S-transferase.
TLDR
The values of Vmax for the intra- and extracellular conjugation reactions differ by 700-fold, suggesting the presence of an intracellular activator for this enzyme system. Expand
Regulation of Thiol Contents in Maize Roots by Intermediates and Effectors of Glutathione Synthesis
Summary The regulation of the synthesis and the levels of GSH and other acid-soluble thiols was studied in maize roots by applying GSH, GSH monoethylester, OTC (L-2-oxothiazolidine-4-carboxylicExpand
Glutathione: biosynthesis, metabolism and relationship to stress tolerance explored in transformed plants
TLDR
Since all functions thus far described for GSH in plants are related to the cysteine moiety of the tripeptide, thesehomologues may exercise similar biochemical roles to c-Key words: Oxidative stress, photorespiration, glutathione, EC-gly. Expand
Glutathione biosynthesis in Arabidopsis trichome cells.
TLDR
The results suggest the important physiological significance of trichome cells related to GSH biosynthesis and their possible role as a sink during detoxification processes. Expand
...
1
2
3
4
5
...