GlutaredoxinV1, Main, Exploration, bibRecord, 000694

Transient light-induced intracellular oxidation revealed by redox biosensor.

Identifieur interne : 000694 ( Main/Exploration ); précédent : 000693; suivant : 000695

Transient light-induced intracellular oxidation revealed by redox biosensor.

Auteurs : Vladimir L. Kolossov [États-Unis] ; Jessica N. Beaudoin ; William P. Hanafin ; Stephen J. Diliberto ; Paul J A. Kenis ; H Rex Gaskins

Source :

Biochemical and biophysical research communications [ 1090-2104 ] ; 2013.

RBID : pubmed:24025674

Descripteurs français

English descriptors

KwdEn :
- Animals (MeSH), Biosensing Techniques (MeSH), CHO Cells (MeSH), Cricetinae (MeSH), Cricetulus (MeSH), Cytoplasm (metabolism), Cytosol (metabolism), Disulfides (metabolism), Glutaredoxins (chemistry), Glutaredoxins (genetics), Glutaredoxins (metabolism), Green Fluorescent Proteins (chemistry), Green Fluorescent Proteins (genetics), Green Fluorescent Proteins (metabolism), Humans (MeSH), Light (MeSH), Oxidation-Reduction (MeSH), Recombinant Fusion Proteins (chemistry), Recombinant Fusion Proteins (genetics), Recombinant Fusion Proteins (metabolism), Sulfhydryl Compounds (metabolism).
MESH :
- chemical , chemistry : Glutaredoxins, Green Fluorescent Proteins, Recombinant Fusion Proteins.
- chemical , genetics : Glutaredoxins, Green Fluorescent Proteins, Recombinant Fusion Proteins.
- chemical , metabolism : Disulfides, Glutaredoxins, Green Fluorescent Proteins, Recombinant Fusion Proteins, Sulfhydryl Compounds.
- metabolism : Cytoplasm, Cytosol.
- Animals, Biosensing Techniques, CHO Cells, Cricetinae, Cricetulus, Humans, Light, Oxidation-Reduction.

Abstract

We have implemented a ratiometric, genetically encoded redox-sensitive green fluorescent protein fused to human glutaredoxin (Grx1-roGFP2) to monitor real time intracellular glutathione redox potentials of mammalian cells. This probe enabled detection of media-dependent oxidation of the cytosol triggered by short wavelength excitation. The transient nature of light-induced oxidation was revealed by time-lapse live cell imaging when time intervals of less than 30s were implemented. In contrast, transient ROS generation was not observed with the parental roGFP2 probe without Grx1, which exhibits slower thiol-disulfide exchange. These data demonstrate that the enhanced sensitivity of the Grx1-roGFP2 fusion protein enables the detection of short-lived ROS in living cells. The superior sensitivity of Grx1-roGFP2, however, also enhances responsiveness to environmental cues introducing a greater likelihood of false positive results during image acquisition.

DOI: 10.1016/j.bbrc.2013.09.011
PubMed: 24025674
PubMed Central: PMC3830497

Affiliations:

Links toward previous steps (curation, corpus...)

to stream Main, to step Corpus: 000707
to stream Main, to step Curation: 000707

Le document en format XML

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<term>Cricetinae (MeSH)</term>
<term>Cricetulus (MeSH)</term>
<term>Cytoplasm (metabolism)</term>
<term>Cytosol (metabolism)</term>
<term>Disulfides (metabolism)</term>
<term>Glutaredoxins (chemistry)</term>
<term>Glutaredoxins (genetics)</term>
<term>Glutaredoxins (metabolism)</term>
<term>Green Fluorescent Proteins (chemistry)</term>
<term>Green Fluorescent Proteins (genetics)</term>
<term>Green Fluorescent Proteins (metabolism)</term>
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<term>Cellules CHO (MeSH)</term>
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<term>Glutarédoxines (métabolisme)</term>
<term>Humains (MeSH)</term>
<term>Lumière (MeSH)</term>
<term>Oxydoréduction (MeSH)</term>
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<term>Protéines de fusion recombinantes (génétique)</term>
<term>Protéines de fusion recombinantes (métabolisme)</term>
<term>Protéines à fluorescence verte (composition chimique)</term>
<term>Protéines à fluorescence verte (génétique)</term>
<term>Protéines à fluorescence verte (métabolisme)</term>
<term>Techniques de biocapteur (MeSH)</term>
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<term>Green Fluorescent Proteins</term>
<term>Recombinant Fusion Proteins</term>
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<term>Glutaredoxins</term>
<term>Green Fluorescent Proteins</term>
<term>Recombinant Fusion Proteins</term>
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<term>Protéines à fluorescence verte</term>
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<term>Cricetulus</term>
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<front><div type="abstract" xml:lang="en">We have implemented a ratiometric, genetically encoded redox-sensitive green fluorescent protein fused to human glutaredoxin (Grx1-roGFP2) to monitor real time intracellular glutathione redox potentials of mammalian cells. This probe enabled detection of media-dependent oxidation of the cytosol triggered by short wavelength excitation. The transient nature of light-induced oxidation was revealed by time-lapse live cell imaging when time intervals of less than 30s were implemented. In contrast, transient ROS generation was not observed with the parental roGFP2 probe without Grx1, which exhibits slower thiol-disulfide exchange. These data demonstrate that the enhanced sensitivity of the Grx1-roGFP2 fusion protein enables the detection of short-lived ROS in living cells. The superior sensitivity of Grx1-roGFP2, however, also enhances responsiveness to environmental cues introducing a greater likelihood of false positive results during image acquisition.</div>
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<Abstract><AbstractText>We have implemented a ratiometric, genetically encoded redox-sensitive green fluorescent protein fused to human glutaredoxin (Grx1-roGFP2) to monitor real time intracellular glutathione redox potentials of mammalian cells. This probe enabled detection of media-dependent oxidation of the cytosol triggered by short wavelength excitation. The transient nature of light-induced oxidation was revealed by time-lapse live cell imaging when time intervals of less than 30s were implemented. In contrast, transient ROS generation was not observed with the parental roGFP2 probe without Grx1, which exhibits slower thiol-disulfide exchange. These data demonstrate that the enhanced sensitivity of the Grx1-roGFP2 fusion protein enables the detection of short-lived ROS in living cells. The superior sensitivity of Grx1-roGFP2, however, also enhances responsiveness to environmental cues introducing a greater likelihood of false positive results during image acquisition.</AbstractText>
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Transient light-induced intracellular oxidation revealed by redox biosensor.

Transient light-induced intracellular oxidation revealed by redox biosensor.

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