- The NADH:ubiquinone oxidoreductase (respiratory complex I) is the main entry point for electrons into the Escherichia coli aerobic respiratory chain. With its sophisticated setup of 13 different subunits and 10 cofactors, it is anticipated that various chaperones are needed for its proper maturation. However, very little is known about the assembly of E. coli complex I, especially concerning the incorporation of the iron-sulfur clusters. To identify iron-sulfur cluster carrier proteins possibly involved in the process, we generated knockout strains of NfuA, BolA, YajL, Mrp, GrxD and IbaG that have been reported either to be involved in the maturation of mitochondrial complex I or to exert influence on the clusters of bacterial complex. We determined the NADH and succinate oxidase activities of membranes from the mutant strains to monitor the specificity of the individual mutations for complex I. The deletion of NfuA, BolA and Mrp led to a decreased stability and partially disturbed assembly of the complex as determined by sucroseThe NADH:ubiquinone oxidoreductase (respiratory complex I) is the main entry point for electrons into the Escherichia coli aerobic respiratory chain. With its sophisticated setup of 13 different subunits and 10 cofactors, it is anticipated that various chaperones are needed for its proper maturation. However, very little is known about the assembly of E. coli complex I, especially concerning the incorporation of the iron-sulfur clusters. To identify iron-sulfur cluster carrier proteins possibly involved in the process, we generated knockout strains of NfuA, BolA, YajL, Mrp, GrxD and IbaG that have been reported either to be involved in the maturation of mitochondrial complex I or to exert influence on the clusters of bacterial complex. We determined the NADH and succinate oxidase activities of membranes from the mutant strains to monitor the specificity of the individual mutations for complex I. The deletion of NfuA, BolA and Mrp led to a decreased stability and partially disturbed assembly of the complex as determined by sucrose gradient centrifugation and native PAGE. EPR spectroscopy of cytoplasmic membranes revealed that the BolA deletion results in the loss of the binuclear Fe/S cluster N1b.…
MetadatenVerfasserangaben: | Sabrina Burschel, Doris Kreuzer Decovic, Franziska Nuber, Marie Stiller, Maud Hofmann, Arkadiusz ZupokORCiDGND, Beata Siemiatkowska, Michal Jakub GorkaGND, Silke LeimkühlerORCiDGND, Thorsten FriedrichORCiD |
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DOI: | https://doi.org/10.1111/mmi.14137 |
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ISSN: | 0950-382X |
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ISSN: | 1365-2958 |
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Pubmed ID: | https://pubmed.ncbi.nlm.nih.gov/30251413 |
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Titel des übergeordneten Werks (Englisch): | Molecular microbiology |
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Untertitel (Englisch): | ubiquinone oxidoreductase (complex I) |
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Verlag: | Wiley |
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Verlagsort: | Hoboken |
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Publikationstyp: | Wissenschaftlicher Artikel |
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Sprache: | Englisch |
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Datum der Erstveröffentlichung: | 24.09.2018 |
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Erscheinungsjahr: | 2018 |
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Datum der Freischaltung: | 25.05.2021 |
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Band: | 111 |
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Ausgabe: | 1 |
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Seitenanzahl: | 15 |
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Erste Seite: | 31 |
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Letzte Seite: | 45 |
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Fördernde Institution: | Deutsche Forschungsgemeinschaft (DFG)German Research Foundation (DFG) [GRK 1976, GRK 2202, SPP 1927 (FR 1140/11-1)]; Spemann Graduate School of Biology and Medicine (SGBM) |
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Organisationseinheiten: | Mathematisch-Naturwissenschaftliche Fakultät / Institut für Biochemie und Biologie |
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DDC-Klassifikation: | 5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie |
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Peer Review: | Referiert |
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Publikationsweg: | Open Access / Bronze Open-Access |
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