Real-time imaging of the intracellular glutathione redox potential.
Identifieur interne : 000B79 ( Main/Exploration ); précédent : 000B78; suivant : 000B80Real-time imaging of the intracellular glutathione redox potential.
Auteurs : Marcus Gutscher [Allemagne] ; Anne-Laure Pauleau ; Laurent Marty ; Thorsten Brach ; Guido H. Wabnitz ; Yvonne Samstag ; Andreas J. Meyer ; Tobias P. DickSource :
- Nature methods [ 1548-7105 ] ; 2008.
Descripteurs français
- KwdFr :
- Apoptose (MeSH), Biotechnologie (méthodes), Cellules HeLa (MeSH), Disulfures (composition chimique), Glutarédoxines (composition chimique), Glutathion (composition chimique), Humains (MeSH), Mitochondries (métabolisme), Oxydoréduction (MeSH), Oxygène (métabolisme), Protéines de fusion recombinantes (composition chimique), Protéines à fluorescence verte (composition chimique), Protéines à fluorescence verte (métabolisme), Sensibilité et spécificité (MeSH), Techniques de biocapteur (MeSH), Thiorédoxines (composition chimique).
- MESH :
- composition chimique : Disulfures, Glutarédoxines, Glutathion, Protéines de fusion recombinantes, Protéines à fluorescence verte, Thiorédoxines.
- métabolisme : Mitochondries, Oxygène, Protéines à fluorescence verte.
- méthodes : Biotechnologie.
- Apoptose, Cellules HeLa, Humains, Oxydoréduction, Sensibilité et spécificité, Techniques de biocapteur.
English descriptors
- KwdEn :
- Apoptosis (MeSH), Biosensing Techniques (MeSH), Biotechnology (methods), Disulfides (chemistry), Glutaredoxins (chemistry), Glutathione (chemistry), Green Fluorescent Proteins (chemistry), Green Fluorescent Proteins (metabolism), HeLa Cells (MeSH), Humans (MeSH), Mitochondria (metabolism), Oxidation-Reduction (MeSH), Oxygen (metabolism), Recombinant Fusion Proteins (chemistry), Sensitivity and Specificity (MeSH), Thioredoxins (chemistry).
- MESH :
- chemical , chemistry : Disulfides, Glutaredoxins, Glutathione, Green Fluorescent Proteins, Recombinant Fusion Proteins, Thioredoxins.
- chemical , metabolism : Green Fluorescent Proteins, Oxygen.
- metabolism : Mitochondria.
- methods : Biotechnology.
- Apoptosis, Biosensing Techniques, HeLa Cells, Humans, Oxidation-Reduction, Sensitivity and Specificity.
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.
DOI: 10.1038/nmeth.1212
PubMed: 18469822
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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Wabnitz</name></author><author><name sortKey="Samstag, Yvonne" sort="Samstag, Yvonne" uniqKey="Samstag Y" first="Yvonne" last="Samstag">Yvonne Samstag</name></author><author><name sortKey="Meyer, Andreas J" sort="Meyer, Andreas J" uniqKey="Meyer A" first="Andreas J" last="Meyer">Andreas J. Meyer</name></author><author><name sortKey="Dick, Tobias P" sort="Dick, Tobias P" uniqKey="Dick T" first="Tobias P" last="Dick">Tobias P. Dick</name></author></analytic><series><title level="j">Nature methods</title><idno type="eISSN">1548-7105</idno><imprint><date when="2008" type="published">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Apoptosis (MeSH)</term><term>Biosensing Techniques (MeSH)</term><term>Biotechnology (methods)</term><term>Disulfides (chemistry)</term><term>Glutaredoxins (chemistry)</term><term>Glutathione (chemistry)</term><term>Green Fluorescent Proteins (chemistry)</term><term>Green Fluorescent Proteins (metabolism)</term><term>HeLa Cells (MeSH)</term><term>Humans (MeSH)</term><term>Mitochondria (metabolism)</term><term>Oxidation-Reduction (MeSH)</term><term>Oxygen (metabolism)</term><term>Recombinant Fusion Proteins (chemistry)</term><term>Sensitivity and Specificity (MeSH)</term><term>Thioredoxins (chemistry)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Apoptose (MeSH)</term><term>Biotechnologie (méthodes)</term><term>Cellules HeLa (MeSH)</term><term>Disulfures (composition chimique)</term><term>Glutarédoxines (composition chimique)</term><term>Glutathion (composition chimique)</term><term>Humains (MeSH)</term><term>Mitochondries (métabolisme)</term><term>Oxydoréduction (MeSH)</term><term>Oxygène (métabolisme)</term><term>Protéines de fusion recombinantes (composition chimique)</term><term>Protéines à fluorescence verte (composition chimique)</term><term>Protéines à fluorescence verte (métabolisme)</term><term>Sensibilité et spécificité (MeSH)</term><term>Techniques de biocapteur (MeSH)</term><term>Thiorédoxines (composition chimique)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Disulfides</term><term>Glutaredoxins</term><term>Glutathione</term><term>Green Fluorescent Proteins</term><term>Recombinant Fusion Proteins</term><term>Thioredoxins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Green Fluorescent Proteins</term><term>Oxygen</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Disulfures</term><term>Glutarédoxines</term><term>Glutathion</term><term>Protéines de fusion recombinantes</term><term>Protéines à fluorescence verte</term><term>Thiorédoxines</term></keywords><keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Mitochondria</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Biotechnology</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Mitochondries</term><term>Oxygène</term><term>Protéines à fluorescence verte</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Biotechnologie</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Apoptosis</term><term>Biosensing Techniques</term><term>HeLa Cells</term><term>Humans</term><term>Oxidation-Reduction</term><term>Sensitivity and Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Apoptose</term><term>Cellules HeLa</term><term>Humains</term><term>Oxydoréduction</term><term>Sensibilité et spécificité</term><term>Techniques de biocapteur</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">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.</div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">18469822</PMID><DateCompleted><Year>2008</Year><Month>06</Month><Day>24</Day></DateCompleted><DateRevised><Year>2013</Year><Month>11</Month><Day>21</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1548-7105</ISSN><JournalIssue CitedMedium="Internet"><Volume>5</Volume><Issue>6</Issue><PubDate><Year>2008</Year><Month>Jun</Month></PubDate></JournalIssue><Title>Nature methods</Title><ISOAbbreviation>Nat Methods</ISOAbbreviation></Journal><ArticleTitle>Real-time imaging of the intracellular glutathione redox potential.</ArticleTitle><Pagination><MedlinePgn>553-9</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1038/nmeth.1212</ELocationID><Abstract><AbstractText>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.</AbstractText></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Gutscher</LastName><ForeName>Marcus</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Redox Regulation Research Group, German Cancer Research Center (DKFZ/A160), Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Pauleau</LastName><ForeName>Anne-Laure</ForeName><Initials>AL</Initials></Author><Author ValidYN="Y"><LastName>Marty</LastName><ForeName>Laurent</ForeName><Initials>L</Initials></Author><Author ValidYN="Y"><LastName>Brach</LastName><ForeName>Thorsten</ForeName><Initials>T</Initials></Author><Author ValidYN="Y"><LastName>Wabnitz</LastName><ForeName>Guido H</ForeName><Initials>GH</Initials></Author><Author ValidYN="Y"><LastName>Samstag</LastName><ForeName>Yvonne</ForeName><Initials>Y</Initials></Author><Author ValidYN="Y"><LastName>Meyer</LastName><ForeName>Andreas J</ForeName><Initials>AJ</Initials></Author><Author ValidYN="Y"><LastName>Dick</LastName><ForeName>Tobias P</ForeName><Initials>TP</Initials></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2008</Year><Month>05</Month><Day>11</Day></ArticleDate></Article><MedlineJournalInfo><Country>United States</Country><MedlineTA>Nat Methods</MedlineTA><NlmUniqueID>101215604</NlmUniqueID><ISSNLinking>1548-7091</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance 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MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006367" MajorTopicYN="N">HeLa Cells</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008928" MajorTopicYN="N">Mitochondria</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010084" MajorTopicYN="Y">Oxidation-Reduction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010100" MajorTopicYN="N">Oxygen</DescriptorName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011993" MajorTopicYN="N">Recombinant Fusion Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D012680" MajorTopicYN="N">Sensitivity and Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013879" MajorTopicYN="N">Thioredoxins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName></MeshHeading></MeshHeadingList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2008</Year><Month>01</Month><Day>15</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2008</Year><Month>04</Month><Day>07</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2008</Year><Month>5</Month><Day>13</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2008</Year><Month>6</Month><Day>25</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2008</Year><Month>5</Month><Day>13</Day><Hour>9</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">18469822</ArticleId><ArticleId IdType="pii">nmeth.1212</ArticleId><ArticleId IdType="doi">10.1038/nmeth.1212</ArticleId></ArticleIdList></PubmedData></pubmed><affiliations><list><country><li>Allemagne</li></country><region><li>Bade-Wurtemberg</li><li>District de Karlsruhe</li></region><settlement><li>Heidelberg</li></settlement></list><tree><noCountry><name sortKey="Brach, Thorsten" sort="Brach, Thorsten" uniqKey="Brach T" first="Thorsten" last="Brach">Thorsten Brach</name><name sortKey="Dick, Tobias P" sort="Dick, Tobias P" uniqKey="Dick T" first="Tobias P" last="Dick">Tobias P. Dick</name><name sortKey="Marty, Laurent" sort="Marty, Laurent" uniqKey="Marty L" first="Laurent" last="Marty">Laurent Marty</name><name sortKey="Meyer, Andreas J" sort="Meyer, Andreas J" uniqKey="Meyer A" first="Andreas J" last="Meyer">Andreas J. Meyer</name><name sortKey="Pauleau, Anne Laure" sort="Pauleau, Anne Laure" uniqKey="Pauleau A" first="Anne-Laure" last="Pauleau">Anne-Laure Pauleau</name><name sortKey="Samstag, Yvonne" sort="Samstag, Yvonne" uniqKey="Samstag Y" first="Yvonne" last="Samstag">Yvonne Samstag</name><name sortKey="Wabnitz, Guido H" sort="Wabnitz, Guido H" uniqKey="Wabnitz G" first="Guido H" last="Wabnitz">Guido H. Wabnitz</name></noCountry><country name="Allemagne"><region name="Bade-Wurtemberg"><name sortKey="Gutscher, Marcus" sort="Gutscher, Marcus" uniqKey="Gutscher M" first="Marcus" last="Gutscher">Marcus Gutscher</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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