Kinetic characterization of wild-type and mutant human thioredoxin glutathione reductase defines its reaction and regulatory mechanisms.
Identifieur interne : 000243 ( Main/Exploration ); précédent : 000242; suivant : 000244Kinetic characterization of wild-type and mutant human thioredoxin glutathione reductase defines its reaction and regulatory mechanisms.
Auteurs : Christina Brandstaedter [Allemagne] ; Karin Fritz-Wolf [Allemagne] ; Stine Weder [Allemagne] ; Marina Fischer [Allemagne] ; Beate Hecker [Allemagne] ; Stefan Rahlfs [Allemagne] ; Katja Becker [Allemagne]Source :
- The FEBS journal [ 1742-4658 ] ; 2018.
Descripteurs français
- KwdFr :
- Biocatalyse (MeSH), Complexes multienzymatiques (composition chimique), Complexes multienzymatiques (génétique), Complexes multienzymatiques (métabolisme), Conformation des protéines (MeSH), Dimérisation (MeSH), Disulfures (composition chimique), Disulfures (métabolisme), Domaine catalytique (MeSH), Fragments peptidiques (composition chimique), Fragments peptidiques (génétique), Fragments peptidiques (métabolisme), Glutathion (composition chimique), Glutathion (métabolisme), Humains (MeSH), Modèles moléculaires (MeSH), Motifs et domaines d'intéraction protéique (MeSH), Mutagenèse dirigée (MeSH), Mutation (MeSH), NADH, NADPH oxidoreductases (composition chimique), NADH, NADPH oxidoreductases (génétique), NADH, NADPH oxidoreductases (métabolisme), Nitro-benzoates (composition chimique), Nitro-benzoates (métabolisme), Oxydoréduction (MeSH), Protéines de fusion recombinantes (composition chimique), Protéines de fusion recombinantes (métabolisme), Sites de fixation (MeSH), Spécificité du substrat (MeSH), Substitution d'acide aminé (MeSH), Thiols (composition chimique), Thiols (métabolisme), Thiorédoxines (composition chimique), Thiorédoxines (métabolisme), Éthanol (analogues et dérivés), Éthanol (composition chimique), Éthanol (métabolisme).
- MESH :
- analogues et dérivés : Éthanol.
- composition chimique : Complexes multienzymatiques, Disulfures, Fragments peptidiques, Glutathion, NADH, NADPH oxidoreductases, Nitro-benzoates, Protéines de fusion recombinantes, Thiols, Thiorédoxines, Éthanol.
- génétique : Complexes multienzymatiques, Fragments peptidiques, NADH, NADPH oxidoreductases.
- métabolisme : Complexes multienzymatiques, Disulfures, Fragments peptidiques, Glutathion, NADH, NADPH oxidoreductases, Nitro-benzoates, Protéines de fusion recombinantes, Thiols, Thiorédoxines, Éthanol.
- Biocatalyse, Conformation des protéines, Dimérisation, Domaine catalytique, Humains, Modèles moléculaires, Motifs et domaines d'intéraction protéique, Mutagenèse dirigée, Mutation, Oxydoréduction, Sites de fixation, Spécificité du substrat, Substitution d'acide aminé.
English descriptors
- KwdEn :
- Amino Acid Substitution (MeSH), Binding Sites (MeSH), Biocatalysis (MeSH), Catalytic Domain (MeSH), Dimerization (MeSH), Disulfides (chemistry), Disulfides (metabolism), Ethanol (analogs & derivatives), Ethanol (chemistry), Ethanol (metabolism), Glutathione (chemistry), Glutathione (metabolism), Humans (MeSH), Models, Molecular (MeSH), Multienzyme Complexes (chemistry), Multienzyme Complexes (genetics), Multienzyme Complexes (metabolism), Mutagenesis, Site-Directed (MeSH), Mutation (MeSH), NADH, NADPH Oxidoreductases (chemistry), NADH, NADPH Oxidoreductases (genetics), NADH, NADPH Oxidoreductases (metabolism), Nitrobenzoates (chemistry), Nitrobenzoates (metabolism), Oxidation-Reduction (MeSH), Peptide Fragments (chemistry), Peptide Fragments (genetics), Peptide Fragments (metabolism), Protein Conformation (MeSH), Protein Interaction Domains and Motifs (MeSH), Recombinant Fusion Proteins (chemistry), Recombinant Fusion Proteins (metabolism), Substrate Specificity (MeSH), Sulfhydryl Compounds (chemistry), Sulfhydryl Compounds (metabolism), Thioredoxins (chemistry), Thioredoxins (metabolism).
- MESH :
- chemical , analogs & derivatives : Ethanol.
- chemical , chemistry : Disulfides, Ethanol, Glutathione, Multienzyme Complexes, NADH, NADPH Oxidoreductases, Nitrobenzoates, Peptide Fragments, Recombinant Fusion Proteins, Sulfhydryl Compounds, Thioredoxins.
- chemical , genetics : Multienzyme Complexes, NADH, NADPH Oxidoreductases, Peptide Fragments.
- chemical , metabolism : Disulfides, Ethanol, Glutathione, Multienzyme Complexes, NADH, NADPH Oxidoreductases, Nitrobenzoates, Peptide Fragments, Recombinant Fusion Proteins, Sulfhydryl Compounds, Thioredoxins.
- Amino Acid Substitution, Binding Sites, Biocatalysis, Catalytic Domain, Dimerization, Humans, Models, Molecular, Mutagenesis, Site-Directed, Mutation, Oxidation-Reduction, Protein Conformation, Protein Interaction Domains and Motifs, Substrate Specificity.
Abstract
In most cells, the thioredoxin (Trx) and glutathione systems are essential in maintaining redox homeostasis. The selenoprotein thioredoxin glutathione reductase (TGR) is a hybrid enzyme in which a glutaredoxin (Grx) domain is linked to a thioredoxin reductase (TrxR). Notably, the protein is also capable of reducing glutathione disulfide (GSSG), thus representing an important link between the two redox systems. In this study, we recombinantly produced human TGR (hTGR wild-type) by fusing its open reading frame with a bacterial selenocysteine insertion sequence element and co-expressing the construct in Escherichia coli together with the selA, selB, and selC genes. Additionally, the Sec→Cys mutant (hTGRU642C ) of the full-length protein, the isolated TrxR domain (hTGR151-643 ) and the Grx domain containing a monothiol active site (hTGR1-150 ) were produced and purified. All four proteins were kinetically characterized in direct comparison using Trx, DTNB, HED, or GSSG as the oxidizing substrate. Interestingly, the HED reduction activity was Sec independent and comparable in the full-length protein and the isolated Grx domain, whereas the TrxR and glutathione reductase reactions were clearly selenocysteine dependent, with the GR reaction requiring the Grx domain. Site-directed mutagenesis studies revealed novel insights into the mechanism of GSSG reduction. Furthermore, we identified several glutathionylation sites in hTGR, including Cys93, Cys133, and Cys619, and an inhibitory effect of these modifications on enzyme activity. In contrast to other TGRs, for example, from platyhelminth parasites, hTGR did not exhibit hysteretic behavior. These findings provide new insights into the reaction mechanism and regulation of monothiol Grx-containing TGRs.
DATABASE
EC numbers: 1.8.1.9; 1.8.1.B1.
DOI: 10.1111/febs.14357
PubMed: 29222842
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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Giessen</wicri:noRegion></affiliation><affiliation wicri:level="3"><nlm:affiliation>Max-Planck Institute for Medical Research, Heidelberg, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Max-Planck Institute for Medical Research, Heidelberg</wicri:regionArea><placeName><region type="land" nuts="1">Bade-Wurtemberg</region><region type="district" nuts="2">District de Karlsruhe</region><settlement type="city">Heidelberg</settlement></placeName></affiliation></author><author><name sortKey="Weder, Stine" sort="Weder, Stine" uniqKey="Weder S" first="Stine" last="Weder">Stine Weder</name><affiliation wicri:level="1"><nlm:affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University 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sort="Hecker, Beate" uniqKey="Hecker B" first="Beate" last="Hecker">Beate Hecker</name><affiliation wicri:level="1"><nlm:affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen</wicri:regionArea><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion></affiliation></author><author><name sortKey="Rahlfs, Stefan" sort="Rahlfs, Stefan" uniqKey="Rahlfs S" first="Stefan" last="Rahlfs">Stefan Rahlfs</name><affiliation wicri:level="1"><nlm:affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen</wicri:regionArea><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion></affiliation></author><author><name sortKey="Becker, Katja" sort="Becker, Katja" uniqKey="Becker K" first="Katja" last="Becker">Katja Becker</name><affiliation wicri:level="1"><nlm:affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</nlm:affiliation><country xml:lang="fr">Allemagne</country><wicri:regionArea>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen</wicri:regionArea><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion><wicri:noRegion>Justus Liebig University Giessen</wicri:noRegion></affiliation></author></analytic><series><title level="j">The FEBS journal</title><idno type="eISSN">1742-4658</idno><imprint><date when="2018" type="published">2018</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Amino Acid Substitution (MeSH)</term><term>Binding Sites (MeSH)</term><term>Biocatalysis (MeSH)</term><term>Catalytic Domain (MeSH)</term><term>Dimerization (MeSH)</term><term>Disulfides (chemistry)</term><term>Disulfides (metabolism)</term><term>Ethanol (analogs & derivatives)</term><term>Ethanol (chemistry)</term><term>Ethanol (metabolism)</term><term>Glutathione (chemistry)</term><term>Glutathione (metabolism)</term><term>Humans (MeSH)</term><term>Models, Molecular (MeSH)</term><term>Multienzyme Complexes (chemistry)</term><term>Multienzyme Complexes (genetics)</term><term>Multienzyme Complexes (metabolism)</term><term>Mutagenesis, Site-Directed (MeSH)</term><term>Mutation (MeSH)</term><term>NADH, NADPH Oxidoreductases (chemistry)</term><term>NADH, NADPH Oxidoreductases (genetics)</term><term>NADH, NADPH Oxidoreductases (metabolism)</term><term>Nitrobenzoates (chemistry)</term><term>Nitrobenzoates (metabolism)</term><term>Oxidation-Reduction (MeSH)</term><term>Peptide Fragments (chemistry)</term><term>Peptide Fragments (genetics)</term><term>Peptide Fragments (metabolism)</term><term>Protein Conformation (MeSH)</term><term>Protein Interaction Domains and Motifs (MeSH)</term><term>Recombinant Fusion Proteins (chemistry)</term><term>Recombinant Fusion Proteins (metabolism)</term><term>Substrate Specificity (MeSH)</term><term>Sulfhydryl Compounds (chemistry)</term><term>Sulfhydryl Compounds (metabolism)</term><term>Thioredoxins (chemistry)</term><term>Thioredoxins (metabolism)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Biocatalyse (MeSH)</term><term>Complexes multienzymatiques (composition chimique)</term><term>Complexes multienzymatiques (génétique)</term><term>Complexes multienzymatiques (métabolisme)</term><term>Conformation des protéines (MeSH)</term><term>Dimérisation (MeSH)</term><term>Disulfures (composition chimique)</term><term>Disulfures (métabolisme)</term><term>Domaine catalytique (MeSH)</term><term>Fragments peptidiques (composition chimique)</term><term>Fragments peptidiques (génétique)</term><term>Fragments peptidiques (métabolisme)</term><term>Glutathion (composition chimique)</term><term>Glutathion (métabolisme)</term><term>Humains (MeSH)</term><term>Modèles moléculaires (MeSH)</term><term>Motifs et domaines d'intéraction protéique (MeSH)</term><term>Mutagenèse dirigée (MeSH)</term><term>Mutation (MeSH)</term><term>NADH, NADPH oxidoreductases (composition chimique)</term><term>NADH, NADPH oxidoreductases (génétique)</term><term>NADH, NADPH oxidoreductases (métabolisme)</term><term>Nitro-benzoates (composition chimique)</term><term>Nitro-benzoates (métabolisme)</term><term>Oxydoréduction (MeSH)</term><term>Protéines de fusion recombinantes (composition chimique)</term><term>Protéines de fusion recombinantes (métabolisme)</term><term>Sites de fixation (MeSH)</term><term>Spécificité du substrat (MeSH)</term><term>Substitution d'acide aminé (MeSH)</term><term>Thiols (composition chimique)</term><term>Thiols (métabolisme)</term><term>Thiorédoxines (composition chimique)</term><term>Thiorédoxines (métabolisme)</term><term>Éthanol (analogues et dérivés)</term><term>Éthanol (composition chimique)</term><term>Éthanol (métabolisme)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analogs & derivatives" xml:lang="en"><term>Ethanol</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>Disulfides</term><term>Ethanol</term><term>Glutathione</term><term>Multienzyme Complexes</term><term>NADH, NADPH Oxidoreductases</term><term>Nitrobenzoates</term><term>Peptide Fragments</term><term>Recombinant Fusion Proteins</term><term>Sulfhydryl Compounds</term><term>Thioredoxins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Multienzyme Complexes</term><term>NADH, NADPH Oxidoreductases</term><term>Peptide Fragments</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Disulfides</term><term>Ethanol</term><term>Glutathione</term><term>Multienzyme Complexes</term><term>NADH, NADPH Oxidoreductases</term><term>Nitrobenzoates</term><term>Peptide Fragments</term><term>Recombinant Fusion Proteins</term><term>Sulfhydryl Compounds</term><term>Thioredoxins</term></keywords><keywords scheme="MESH" qualifier="analogues et dérivés" xml:lang="fr"><term>Éthanol</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Complexes multienzymatiques</term><term>Disulfures</term><term>Fragments peptidiques</term><term>Glutathion</term><term>NADH, NADPH oxidoreductases</term><term>Nitro-benzoates</term><term>Protéines de fusion recombinantes</term><term>Thiols</term><term>Thiorédoxines</term><term>Éthanol</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Complexes multienzymatiques</term><term>Fragments peptidiques</term><term>NADH, NADPH oxidoreductases</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Complexes multienzymatiques</term><term>Disulfures</term><term>Fragments peptidiques</term><term>Glutathion</term><term>NADH, NADPH oxidoreductases</term><term>Nitro-benzoates</term><term>Protéines de fusion recombinantes</term><term>Thiols</term><term>Thiorédoxines</term><term>Éthanol</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Amino Acid Substitution</term><term>Binding Sites</term><term>Biocatalysis</term><term>Catalytic Domain</term><term>Dimerization</term><term>Humans</term><term>Models, Molecular</term><term>Mutagenesis, Site-Directed</term><term>Mutation</term><term>Oxidation-Reduction</term><term>Protein Conformation</term><term>Protein Interaction Domains and Motifs</term><term>Substrate Specificity</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Biocatalyse</term><term>Conformation des protéines</term><term>Dimérisation</term><term>Domaine catalytique</term><term>Humains</term><term>Modèles moléculaires</term><term>Motifs et domaines d'intéraction protéique</term><term>Mutagenèse dirigée</term><term>Mutation</term><term>Oxydoréduction</term><term>Sites de fixation</term><term>Spécificité du substrat</term><term>Substitution d'acide aminé</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">In most cells, the thioredoxin (Trx) and glutathione systems are essential in maintaining redox homeostasis. The selenoprotein thioredoxin glutathione reductase (TGR) is a hybrid enzyme in which a glutaredoxin (Grx) domain is linked to a thioredoxin reductase (TrxR). Notably, the protein is also capable of reducing glutathione disulfide (GSSG), thus representing an important link between the two redox systems. In this study, we recombinantly produced human TGR (hTGR wild-type) by fusing its open reading frame with a bacterial selenocysteine insertion sequence element and co-expressing the construct in Escherichia coli together with the selA, selB, and selC genes. Additionally, the Sec→Cys mutant (hTGR<sup>U</sup><sup>642C</sup> ) of the full-length protein, the isolated TrxR domain (hTGR<sup>151-643</sup> ) and the Grx domain containing a monothiol active site (hTGR<sup>1-150</sup> ) were produced and purified. All four proteins were kinetically characterized in direct comparison using Trx, DTNB, HED, or GSSG as the oxidizing substrate. Interestingly, the HED reduction activity was Sec independent and comparable in the full-length protein and the isolated Grx domain, whereas the TrxR and glutathione reductase reactions were clearly selenocysteine dependent, with the GR reaction requiring the Grx domain. Site-directed mutagenesis studies revealed novel insights into the mechanism of GSSG reduction. Furthermore, we identified several glutathionylation sites in hTGR, including Cys93, Cys133, and Cys619, and an inhibitory effect of these modifications on enzyme activity. In contrast to other TGRs, for example, from platyhelminth parasites, hTGR did not exhibit hysteretic behavior. These findings provide new insights into the reaction mechanism and regulation of monothiol Grx-containing TGRs.</div><div type="abstract" xml:lang="en"><p><b>DATABASE</b></p><p>EC numbers: 1.8.1.9; 1.8.1.B1.</p></div></front></TEI><pubmed><MedlineCitation Status="MEDLINE" Owner="NLM"><PMID Version="1">29222842</PMID><DateCompleted><Year>2018</Year><Month>12</Month><Day>11</Day></DateCompleted><DateRevised><Year>2018</Year><Month>12</Month><Day>11</Day></DateRevised><Article PubModel="Print-Electronic"><Journal><ISSN IssnType="Electronic">1742-4658</ISSN><JournalIssue CitedMedium="Internet"><Volume>285</Volume><Issue>3</Issue><PubDate><Year>2018</Year><Month>02</Month></PubDate></JournalIssue><Title>The FEBS journal</Title><ISOAbbreviation>FEBS J</ISOAbbreviation></Journal><ArticleTitle>Kinetic characterization of wild-type and mutant human thioredoxin glutathione reductase defines its reaction and regulatory mechanisms.</ArticleTitle><Pagination><MedlinePgn>542-558</MedlinePgn></Pagination><ELocationID EIdType="doi" ValidYN="Y">10.1111/febs.14357</ELocationID><Abstract><AbstractText>In most cells, the thioredoxin (Trx) and glutathione systems are essential in maintaining redox homeostasis. The selenoprotein thioredoxin glutathione reductase (TGR) is a hybrid enzyme in which a glutaredoxin (Grx) domain is linked to a thioredoxin reductase (TrxR). Notably, the protein is also capable of reducing glutathione disulfide (GSSG), thus representing an important link between the two redox systems. In this study, we recombinantly produced human TGR (hTGR wild-type) by fusing its open reading frame with a bacterial selenocysteine insertion sequence element and co-expressing the construct in Escherichia coli together with the selA, selB, and selC genes. Additionally, the Sec→Cys mutant (hTGR<sup>U</sup><sup>642C</sup> ) of the full-length protein, the isolated TrxR domain (hTGR<sup>151-643</sup> ) and the Grx domain containing a monothiol active site (hTGR<sup>1-150</sup> ) were produced and purified. All four proteins were kinetically characterized in direct comparison using Trx, DTNB, HED, or GSSG as the oxidizing substrate. Interestingly, the HED reduction activity was Sec independent and comparable in the full-length protein and the isolated Grx domain, whereas the TrxR and glutathione reductase reactions were clearly selenocysteine dependent, with the GR reaction requiring the Grx domain. Site-directed mutagenesis studies revealed novel insights into the mechanism of GSSG reduction. Furthermore, we identified several glutathionylation sites in hTGR, including Cys93, Cys133, and Cys619, and an inhibitory effect of these modifications on enzyme activity. In contrast to other TGRs, for example, from platyhelminth parasites, hTGR did not exhibit hysteretic behavior. These findings provide new insights into the reaction mechanism and regulation of monothiol Grx-containing TGRs.</AbstractText><AbstractText Label="DATABASE">EC numbers: 1.8.1.9; 1.8.1.B1.</AbstractText><CopyrightInformation>© 2017 Federation of European Biochemical Societies.</CopyrightInformation></Abstract><AuthorList CompleteYN="Y"><Author ValidYN="Y"><LastName>Brandstaedter</LastName><ForeName>Christina</ForeName><Initials>C</Initials><AffiliationInfo><Affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fritz-Wolf</LastName><ForeName>Karin</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</Affiliation></AffiliationInfo><AffiliationInfo><Affiliation>Max-Planck Institute for Medical Research, Heidelberg, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Weder</LastName><ForeName>Stine</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Fischer</LastName><ForeName>Marina</ForeName><Initials>M</Initials><AffiliationInfo><Affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Hecker</LastName><ForeName>Beate</ForeName><Initials>B</Initials><AffiliationInfo><Affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Rahlfs</LastName><ForeName>Stefan</ForeName><Initials>S</Initials><AffiliationInfo><Affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</Affiliation></AffiliationInfo></Author><Author ValidYN="Y"><LastName>Becker</LastName><ForeName>Katja</ForeName><Initials>K</Initials><AffiliationInfo><Affiliation>Biochemistry and Molecular Biology, Interdisciplinary Research Center, Justus Liebig University Giessen, Germany.</Affiliation></AffiliationInfo></Author></AuthorList><Language>eng</Language><PublicationTypeList><PublicationType UI="D003160">Comparative Study</PublicationType><PublicationType UI="D016428">Journal Article</PublicationType><PublicationType UI="D013485">Research Support, Non-U.S. Gov't</PublicationType></PublicationTypeList><ArticleDate DateType="Electronic"><Year>2017</Year><Month>12</Month><Day>29</Day></ArticleDate></Article><MedlineJournalInfo><Country>England</Country><MedlineTA>FEBS J</MedlineTA><NlmUniqueID>101229646</NlmUniqueID><ISSNLinking>1742-464X</ISSNLinking></MedlineJournalInfo><ChemicalList><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D004220">Disulfides</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009097">Multienzyme Complexes</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D009579">Nitrobenzoates</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D010446">Peptide Fragments</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D011993">Recombinant Fusion Proteins</NameOfSubstance></Chemical><Chemical><RegistryNumber>0</RegistryNumber><NameOfSubstance UI="D013438">Sulfhydryl Compounds</NameOfSubstance></Chemical><Chemical><RegistryNumber>15139-21-6</RegistryNumber><NameOfSubstance UI="C011136">thionitrobenzoic acid</NameOfSubstance></Chemical><Chemical><RegistryNumber>3K9958V90M</RegistryNumber><NameOfSubstance UI="D000431">Ethanol</NameOfSubstance></Chemical><Chemical><RegistryNumber>45543L74BS</RegistryNumber><NameOfSubstance UI="C031319">2-hydroxyethyl disulfide</NameOfSubstance></Chemical><Chemical><RegistryNumber>52500-60-4</RegistryNumber><NameOfSubstance UI="D013879">Thioredoxins</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.6.-</RegistryNumber><NameOfSubstance UI="D009247">NADH, NADPH Oxidoreductases</NameOfSubstance></Chemical><Chemical><RegistryNumber>EC 1.6.4.-</RegistryNumber><NameOfSubstance UI="C466433">thioredoxin glutathione reductase</NameOfSubstance></Chemical><Chemical><RegistryNumber>GAN16C9B8O</RegistryNumber><NameOfSubstance UI="D005978">Glutathione</NameOfSubstance></Chemical></ChemicalList><CitationSubset>IM</CitationSubset><MeshHeadingList><MeshHeading><DescriptorName UI="D019943" MajorTopicYN="N">Amino Acid Substitution</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D001665" MajorTopicYN="N">Binding Sites</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D055162" MajorTopicYN="N">Biocatalysis</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D020134" MajorTopicYN="N">Catalytic Domain</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D019281" MajorTopicYN="N">Dimerization</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D004220" MajorTopicYN="N">Disulfides</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D000431" MajorTopicYN="N">Ethanol</DescriptorName><QualifierName UI="Q000031" MajorTopicYN="N">analogs & derivatives</QualifierName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D005978" MajorTopicYN="N">Glutathione</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D006801" MajorTopicYN="N">Humans</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D008958" MajorTopicYN="Y">Models, Molecular</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009097" MajorTopicYN="N">Multienzyme Complexes</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D016297" MajorTopicYN="N">Mutagenesis, Site-Directed</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009154" MajorTopicYN="N">Mutation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D009247" MajorTopicYN="N">NADH, NADPH Oxidoreductases</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D009579" MajorTopicYN="N">Nitrobenzoates</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D010084" MajorTopicYN="N">Oxidation-Reduction</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D010446" MajorTopicYN="N">Peptide Fragments</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000235" MajorTopicYN="N">genetics</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D011487" MajorTopicYN="N">Protein Conformation</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D054730" MajorTopicYN="N">Protein Interaction Domains and Motifs</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D011993" MajorTopicYN="N">Recombinant Fusion Proteins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013379" MajorTopicYN="N">Substrate Specificity</DescriptorName></MeshHeading><MeshHeading><DescriptorName UI="D013438" MajorTopicYN="N">Sulfhydryl Compounds</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="N">metabolism</QualifierName></MeshHeading><MeshHeading><DescriptorName UI="D013879" MajorTopicYN="N">Thioredoxins</DescriptorName><QualifierName UI="Q000737" MajorTopicYN="N">chemistry</QualifierName><QualifierName UI="Q000378" MajorTopicYN="Y">metabolism</QualifierName></MeshHeading></MeshHeadingList><KeywordList Owner="NOTNLM"><Keyword MajorTopicYN="Y">disulfide reductase</Keyword><Keyword MajorTopicYN="Y">flavoproteins</Keyword><Keyword MajorTopicYN="Y">glutaredoxin</Keyword><Keyword MajorTopicYN="Y">redox metabolism</Keyword><Keyword MajorTopicYN="Y">selenoprotein</Keyword></KeywordList></MedlineCitation><PubmedData><History><PubMedPubDate PubStatus="received"><Year>2017</Year><Month>07</Month><Day>31</Day></PubMedPubDate><PubMedPubDate PubStatus="revised"><Year>2017</Year><Month>11</Month><Day>20</Day></PubMedPubDate><PubMedPubDate PubStatus="accepted"><Year>2017</Year><Month>12</Month><Day>05</Day></PubMedPubDate><PubMedPubDate PubStatus="pubmed"><Year>2017</Year><Month>12</Month><Day>10</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="medline"><Year>2018</Year><Month>12</Month><Day>12</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate><PubMedPubDate PubStatus="entrez"><Year>2017</Year><Month>12</Month><Day>10</Day><Hour>6</Hour><Minute>0</Minute></PubMedPubDate></History><PublicationStatus>ppublish</PublicationStatus><ArticleIdList><ArticleId IdType="pubmed">29222842</ArticleId><ArticleId IdType="doi">10.1111/febs.14357</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><country name="Allemagne"><noRegion><name sortKey="Brandstaedter, Christina" sort="Brandstaedter, Christina" uniqKey="Brandstaedter C" first="Christina" last="Brandstaedter">Christina Brandstaedter</name></noRegion><name sortKey="Becker, Katja" sort="Becker, Katja" uniqKey="Becker K" first="Katja" last="Becker">Katja Becker</name><name sortKey="Fischer, Marina" sort="Fischer, Marina" uniqKey="Fischer M" first="Marina" last="Fischer">Marina Fischer</name><name sortKey="Fritz Wolf, Karin" sort="Fritz Wolf, Karin" uniqKey="Fritz Wolf K" first="Karin" last="Fritz-Wolf">Karin Fritz-Wolf</name><name sortKey="Fritz Wolf, Karin" sort="Fritz Wolf, Karin" uniqKey="Fritz Wolf K" first="Karin" last="Fritz-Wolf">Karin Fritz-Wolf</name><name sortKey="Hecker, Beate" sort="Hecker, Beate" uniqKey="Hecker B" first="Beate" last="Hecker">Beate Hecker</name><name sortKey="Rahlfs, Stefan" sort="Rahlfs, Stefan" uniqKey="Rahlfs S" first="Stefan" last="Rahlfs">Stefan Rahlfs</name><name sortKey="Weder, Stine" sort="Weder, Stine" uniqKey="Weder S" first="Stine" last="Weder">Stine Weder</name></country></tree></affiliations></record>Pour 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