Red fluorescent redox-sensitive biosensor Grx1-roCherry.
Identifieur interne : 000128 ( Main/Exploration ); précédent : 000127; suivant : 000129Red fluorescent redox-sensitive biosensor Grx1-roCherry.
Auteurs : Arina G. Shokhina [Russie] ; Alexander I. Kostyuk [Russie] ; Yulia G. Ermakova [Allemagne] ; Anastasiya S. Panova [Russie] ; Dmitry B. Staroverov [Russie] ; Evgeny S. Egorov [Russie] ; Mikhail S. Baranov [Russie] ; Gijsbert J. Van Belle [Allemagne] ; Dörthe M. Katschinski [Allemagne] ; Vsevolod V. Belousov [Russie] ; Dmitry S. Bilan [Russie]Source :
- Redox biology [ 2213-2317 ] ; 2019.
Descripteurs français
- KwdFr :
- Animaux (MeSH), Cellules HEK293 (MeSH), Cellules HeLa (MeSH), Danio zébré (MeSH), Disulfure de glutathion (métabolisme), Expression des gènes (MeSH), Glutarédoxines (génétique), Glutarédoxines (métabolisme), Glutathion (métabolisme), Glycolyse (MeSH), Gènes rapporteurs (MeSH), Humains (MeSH), Hypoxie (métabolisme), Oxydoréduction (MeSH), Protéines de fusion recombinantes (MeSH), Protéines luminescentes (génétique), Protéines luminescentes (métabolisme), Souris (MeSH), Stress oxydatif (MeSH), Techniques de biocapteur (MeSH).
- MESH :
- génétique : Glutarédoxines, Protéines luminescentes.
- métabolisme : Disulfure de glutathion, Glutarédoxines, Glutathion, Hypoxie, Protéines luminescentes.
- Animaux, Cellules HEK293, Cellules HeLa, Danio zébré, Expression des gènes, Glycolyse, Gènes rapporteurs, Humains, Oxydoréduction, Protéines de fusion recombinantes, Souris, Stress oxydatif, Techniques de biocapteur.
English descriptors
- KwdEn :
- Animals (MeSH), Biosensing Techniques (MeSH), Gene Expression (MeSH), Genes, Reporter (MeSH), Glutaredoxins (genetics), Glutaredoxins (metabolism), Glutathione (metabolism), Glutathione Disulfide (metabolism), Glycolysis (MeSH), HEK293 Cells (MeSH), HeLa Cells (MeSH), Humans (MeSH), Hypoxia (metabolism), Luminescent Proteins (genetics), Luminescent Proteins (metabolism), Mice (MeSH), Oxidation-Reduction (MeSH), Oxidative Stress (MeSH), Recombinant Fusion Proteins (MeSH), Zebrafish (MeSH).
- MESH :
- chemical , genetics : Glutaredoxins, Luminescent Proteins.
- chemical , metabolism : Glutaredoxins, Glutathione, Glutathione Disulfide, Luminescent Proteins.
- metabolism : Hypoxia.
- Animals, Biosensing Techniques, Gene Expression, Genes, Reporter, Glycolysis, HEK293 Cells, HeLa Cells, Humans, Mice, Oxidation-Reduction, Oxidative Stress, Recombinant Fusion Proteins, Zebrafish.
Abstract
Redox-sensitive fluorescent proteins (roFPs) are a powerful tool for imaging intracellular redox changes. The structure of these proteins contains a pair of cysteines capable of forming a disulfide upon oxidation that affects the protein conformation and spectral characteristics. To date, a palette of such biosensors covers the spectral range from blue to red. However, most of the roFPs suffer from either poor brightness or high pH-dependency, or both. Moreover, there is no roRFP with the redox potential close to that of 2GSH/GSSG redox pair. In the present work, we describe Grx1-roCherry, the first red roFP with canonical FP topology and fluorescent excitation/emission spectra of typical RFP. Grx1-roCherry, with a midpoint redox potential of - 311 mV, is characterized by high brightness and increased pH stability (pKa 6.7). We successfully used Grx1-roCherry in combination with other biosensors in a multiparameter imaging mode to demonstrate redox changes in cells under various metabolic perturbations, including hypoxia/reoxygenation. In particular, using simultaneous expression of Grx1-roCherry and its green analog in various compartments of living cells, we demonstrated that local H2O2 production leads to compartment-specific and cell-type-specific changes in the 2GSH/GSSG ratio. Finally, we demonstrate the utility of Grx1-roCherry for in vivo redox imaging.
DOI: 10.1016/j.redox.2018.101071
PubMed: 30576927
PubMed Central: PMC6302151
Affiliations:
- Allemagne, Russie
- Bade-Wurtemberg, Basse-Saxe, District de Karlsruhe, District fédéral central
- Göttingen, Heidelberg, Moscou
Links toward previous steps (curation, corpus...)
Le document en format XML
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<author><name sortKey="Egorov, Evgeny S" sort="Egorov, Evgeny S" uniqKey="Egorov E" first="Evgeny S" last="Egorov">Evgeny S. Egorov</name>
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<author><name sortKey="Baranov, Mikhail S" sort="Baranov, Mikhail S" uniqKey="Baranov M" first="Mikhail S" last="Baranov">Mikhail S. Baranov</name>
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<author><name sortKey="Van Belle, Gijsbert J" sort="Van Belle, Gijsbert J" uniqKey="Van Belle G" first="Gijsbert J" last="Van Belle">Gijsbert J. Van Belle</name>
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<author><name sortKey="Katschinski, Dorthe M" sort="Katschinski, Dorthe M" uniqKey="Katschinski D" first="Dörthe M" last="Katschinski">Dörthe M. Katschinski</name>
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<country xml:lang="fr">Allemagne</country>
<wicri:regionArea>Institute for Cardiovascular Physiology, Georg August University Göttingen, Göttingen 37073</wicri:regionArea>
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<author><name sortKey="Belousov, Vsevolod V" sort="Belousov, Vsevolod V" uniqKey="Belousov V" first="Vsevolod V" last="Belousov">Vsevolod V. Belousov</name>
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<country xml:lang="fr">Russie</country>
<wicri:regionArea>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia; Institute for Cardiovascular Physiology, Georg August University Göttingen, Göttingen 37073, Germany; Pirogov Russian National Research Medical University, Moscow 117997</wicri:regionArea>
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<author><name sortKey="Bilan, Dmitry S" sort="Bilan, Dmitry S" uniqKey="Bilan D" first="Dmitry S" last="Bilan">Dmitry S. Bilan</name>
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals (MeSH)</term>
<term>Biosensing Techniques (MeSH)</term>
<term>Gene Expression (MeSH)</term>
<term>Genes, Reporter (MeSH)</term>
<term>Glutaredoxins (genetics)</term>
<term>Glutaredoxins (metabolism)</term>
<term>Glutathione (metabolism)</term>
<term>Glutathione Disulfide (metabolism)</term>
<term>Glycolysis (MeSH)</term>
<term>HEK293 Cells (MeSH)</term>
<term>HeLa Cells (MeSH)</term>
<term>Humans (MeSH)</term>
<term>Hypoxia (metabolism)</term>
<term>Luminescent Proteins (genetics)</term>
<term>Luminescent Proteins (metabolism)</term>
<term>Mice (MeSH)</term>
<term>Oxidation-Reduction (MeSH)</term>
<term>Oxidative Stress (MeSH)</term>
<term>Recombinant Fusion Proteins (MeSH)</term>
<term>Zebrafish (MeSH)</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux (MeSH)</term>
<term>Cellules HEK293 (MeSH)</term>
<term>Cellules HeLa (MeSH)</term>
<term>Danio zébré (MeSH)</term>
<term>Disulfure de glutathion (métabolisme)</term>
<term>Expression des gènes (MeSH)</term>
<term>Glutarédoxines (génétique)</term>
<term>Glutarédoxines (métabolisme)</term>
<term>Glutathion (métabolisme)</term>
<term>Glycolyse (MeSH)</term>
<term>Gènes rapporteurs (MeSH)</term>
<term>Humains (MeSH)</term>
<term>Hypoxie (métabolisme)</term>
<term>Oxydoréduction (MeSH)</term>
<term>Protéines de fusion recombinantes (MeSH)</term>
<term>Protéines luminescentes (génétique)</term>
<term>Protéines luminescentes (métabolisme)</term>
<term>Souris (MeSH)</term>
<term>Stress oxydatif (MeSH)</term>
<term>Techniques de biocapteur (MeSH)</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Glutaredoxins</term>
<term>Luminescent Proteins</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Glutaredoxins</term>
<term>Glutathione</term>
<term>Glutathione Disulfide</term>
<term>Luminescent Proteins</term>
</keywords>
<keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Glutarédoxines</term>
<term>Protéines luminescentes</term>
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<keywords scheme="MESH" qualifier="metabolism" xml:lang="en"><term>Hypoxia</term>
</keywords>
<keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Disulfure de glutathion</term>
<term>Glutarédoxines</term>
<term>Glutathion</term>
<term>Hypoxie</term>
<term>Protéines luminescentes</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Biosensing Techniques</term>
<term>Gene Expression</term>
<term>Genes, Reporter</term>
<term>Glycolysis</term>
<term>HEK293 Cells</term>
<term>HeLa Cells</term>
<term>Humans</term>
<term>Mice</term>
<term>Oxidation-Reduction</term>
<term>Oxidative Stress</term>
<term>Recombinant Fusion Proteins</term>
<term>Zebrafish</term>
</keywords>
<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Cellules HEK293</term>
<term>Cellules HeLa</term>
<term>Danio zébré</term>
<term>Expression des gènes</term>
<term>Glycolyse</term>
<term>Gènes rapporteurs</term>
<term>Humains</term>
<term>Oxydoréduction</term>
<term>Protéines de fusion recombinantes</term>
<term>Souris</term>
<term>Stress oxydatif</term>
<term>Techniques de biocapteur</term>
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<front><div type="abstract" xml:lang="en">Redox-sensitive fluorescent proteins (roFPs) are a powerful tool for imaging intracellular redox changes. The structure of these proteins contains a pair of cysteines capable of forming a disulfide upon oxidation that affects the protein conformation and spectral characteristics. To date, a palette of such biosensors covers the spectral range from blue to red. However, most of the roFPs suffer from either poor brightness or high pH-dependency, or both. Moreover, there is no roRFP with the redox potential close to that of 2GSH/GSSG redox pair. In the present work, we describe Grx1-roCherry, the first red roFP with canonical FP topology and fluorescent excitation/emission spectra of typical RFP. Grx1-roCherry, with a midpoint redox potential of - 311 mV, is characterized by high brightness and increased pH stability (pKa 6.7). We successfully used Grx1-roCherry in combination with other biosensors in a multiparameter imaging mode to demonstrate redox changes in cells under various metabolic perturbations, including hypoxia/reoxygenation. In particular, using simultaneous expression of Grx1-roCherry and its green analog in various compartments of living cells, we demonstrated that local H<sub>2</sub>
O<sub>2</sub>
production leads to compartment-specific and cell-type-specific changes in the 2GSH/GSSG ratio. Finally, we demonstrate the utility of Grx1-roCherry for in vivo redox imaging.</div>
</front>
</TEI>
<pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">30576927</PMID>
<DateCompleted><Year>2019</Year>
<Month>04</Month>
<Day>03</Day>
</DateCompleted>
<DateRevised><Year>2020</Year>
<Month>03</Month>
<Day>09</Day>
</DateRevised>
<Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">2213-2317</ISSN>
<JournalIssue CitedMedium="Internet"><Volume>21</Volume>
<PubDate><Year>2019</Year>
<Month>02</Month>
</PubDate>
</JournalIssue>
<Title>Redox biology</Title>
<ISOAbbreviation>Redox Biol</ISOAbbreviation>
</Journal>
<ArticleTitle>Red fluorescent redox-sensitive biosensor Grx1-roCherry.</ArticleTitle>
<Pagination><MedlinePgn>101071</MedlinePgn>
</Pagination>
<ELocationID EIdType="pii" ValidYN="Y">S2213-2317(18)30934-0</ELocationID>
<ELocationID EIdType="doi" ValidYN="Y">10.1016/j.redox.2018.101071</ELocationID>
<Abstract><AbstractText>Redox-sensitive fluorescent proteins (roFPs) are a powerful tool for imaging intracellular redox changes. The structure of these proteins contains a pair of cysteines capable of forming a disulfide upon oxidation that affects the protein conformation and spectral characteristics. To date, a palette of such biosensors covers the spectral range from blue to red. However, most of the roFPs suffer from either poor brightness or high pH-dependency, or both. Moreover, there is no roRFP with the redox potential close to that of 2GSH/GSSG redox pair. In the present work, we describe Grx1-roCherry, the first red roFP with canonical FP topology and fluorescent excitation/emission spectra of typical RFP. Grx1-roCherry, with a midpoint redox potential of - 311 mV, is characterized by high brightness and increased pH stability (pKa 6.7). We successfully used Grx1-roCherry in combination with other biosensors in a multiparameter imaging mode to demonstrate redox changes in cells under various metabolic perturbations, including hypoxia/reoxygenation. In particular, using simultaneous expression of Grx1-roCherry and its green analog in various compartments of living cells, we demonstrated that local H<sub>2</sub>
O<sub>2</sub>
production leads to compartment-specific and cell-type-specific changes in the 2GSH/GSSG ratio. Finally, we demonstrate the utility of Grx1-roCherry for in vivo redox imaging.</AbstractText>
<CopyrightInformation>Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.</CopyrightInformation>
</Abstract>
<AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Shokhina</LastName>
<ForeName>Arina G</ForeName>
<Initials>AG</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Kostyuk</LastName>
<ForeName>Alexander I</ForeName>
<Initials>AI</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>Ermakova</LastName>
<ForeName>Yulia G</ForeName>
<Initials>YG</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia; European Molecular Biology Laboratory, Heidelberg 69117, Germany.</Affiliation>
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<Author ValidYN="Y"><LastName>Panova</LastName>
<ForeName>Anastasiya S</ForeName>
<Initials>AS</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
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<ForeName>Dmitry B</ForeName>
<Initials>DB</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
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<ForeName>Evgeny S</ForeName>
<Initials>ES</Initials>
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</AffiliationInfo>
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<ForeName>Mikhail S</ForeName>
<Initials>MS</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
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<Author ValidYN="Y"><LastName>van Belle</LastName>
<ForeName>Gijsbert J</ForeName>
<Initials>GJ</Initials>
<AffiliationInfo><Affiliation>Institute for Cardiovascular Physiology, Georg August University Göttingen, Göttingen 37073, Germany.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Katschinski</LastName>
<ForeName>Dörthe M</ForeName>
<Initials>DM</Initials>
<AffiliationInfo><Affiliation>Institute for Cardiovascular Physiology, Georg August University Göttingen, Göttingen 37073, Germany.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Belousov</LastName>
<ForeName>Vsevolod V</ForeName>
<Initials>VV</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia; Institute for Cardiovascular Physiology, Georg August University Göttingen, Göttingen 37073, Germany; Pirogov Russian National Research Medical University, Moscow 117997, Russia. Electronic address: belousov@ibch.ru.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y"><LastName>Bilan</LastName>
<ForeName>Dmitry S</ForeName>
<Initials>DS</Initials>
<AffiliationInfo><Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia; Pirogov Russian National Research Medical University, Moscow 117997, Russia. Electronic address: d.s.bilan@gmail.com.</Affiliation>
</AffiliationInfo>
</Author>
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<Language>eng</Language>
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<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
</PublicationTypeList>
<ArticleDate DateType="Electronic"><Year>2018</Year>
<Month>12</Month>
<Day>07</Day>
</ArticleDate>
</Article>
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<MedlineTA>Redox Biol</MedlineTA>
<NlmUniqueID>101605639</NlmUniqueID>
<ISSNLinking>2213-2317</ISSNLinking>
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</MeshHeading>
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</MeshHeading>
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<Keyword MajorTopicYN="Y">Biosensor</Keyword>
<Keyword MajorTopicYN="Y">Grx1-roCherry</Keyword>
<Keyword MajorTopicYN="Y">Multiparameter imaging</Keyword>
<Keyword MajorTopicYN="Y">Redox-sensitive fluorescent protein</Keyword>
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<name sortKey="Katschinski, Dorthe M" sort="Katschinski, Dorthe M" uniqKey="Katschinski D" first="Dörthe M" last="Katschinski">Dörthe M. Katschinski</name>
<name sortKey="Van Belle, Gijsbert J" sort="Van Belle, Gijsbert J" uniqKey="Van Belle G" first="Gijsbert J" last="Van Belle">Gijsbert J. Van Belle</name>
</country>
</tree>
</affiliations>
</record>
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