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Red fluorescent redox-sensitive biosensor Grx1-roCherry.

Identifieur interne : 000128 ( Main/Exploration ); précédent : 000127; suivant : 000129

Red 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 :

RBID : pubmed:30576927

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English descriptors

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


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<term>Glutaredoxins (metabolism)</term>
<term>Glutathione (metabolism)</term>
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<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>
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<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>
</Author>
<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>
</AffiliationInfo>
</Author>
<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>
</Author>
<Author ValidYN="Y">
<LastName>Staroverov</LastName>
<ForeName>Dmitry B</ForeName>
<Initials>DB</Initials>
<AffiliationInfo>
<Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
</Author>
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<LastName>Egorov</LastName>
<ForeName>Evgeny S</ForeName>
<Initials>ES</Initials>
<AffiliationInfo>
<Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
</Author>
<Author ValidYN="Y">
<LastName>Baranov</LastName>
<ForeName>Mikhail S</ForeName>
<Initials>MS</Initials>
<AffiliationInfo>
<Affiliation>Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Moscow 117997, Russia.</Affiliation>
</AffiliationInfo>
</Author>
<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>
</AuthorList>
<Language>eng</Language>
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<PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType>
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<ArticleDate DateType="Electronic">
<Year>2018</Year>
<Month>12</Month>
<Day>07</Day>
</ArticleDate>
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<Country>Netherlands</Country>
<MedlineTA>Redox Biol</MedlineTA>
<NlmUniqueID>101605639</NlmUniqueID>
<ISSNLinking>2213-2317</ISSNLinking>
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<DescriptorName UI="D000860" MajorTopicYN="N">Hypoxia</DescriptorName>
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<DescriptorName UI="D051379" MajorTopicYN="N">Mice</DescriptorName>
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<name sortKey="Ermakova, Yulia G" sort="Ermakova, Yulia G" uniqKey="Ermakova Y" first="Yulia G" last="Ermakova">Yulia G. Ermakova</name>
</region>
<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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