caa a22 4500 378896040 CHVBK 20180305123506.0 cr unu---uuuuu 161128e20041201xx s 000 0 eng 10.1515/BC.2004.153 doi (NATIONALLICENCE)gruyter-10.1515/BC.2004.153 Novel thioredoxin targets in Dictyostelium discoideum identified by two-hybrid analysis: interactions of thioredoxin with elongation factor 1α and yeast alcohol dehydrogenase [Elektronische Daten] [Thomas Brodegger, Anja Stockmann, Jürgen Oberstraß, Wolfgang Nellen, Hartmut Follmann] Thioredoxins (Trx) are ubiquitous dicysteine proteins capable of modulating enzymes and other cellular targets through specific disulfide-dithiol redox changes. They are unique in that a large number of very diverse metabolic systems are addressed and redox-regulated in bacteria, animal, and plant cells, but the finite number of thioredoxin interaction partners is still unknown. Two-hybrid methodology should provide a rational way to establish thioredoxin functions in a given organism. We report a search for physiological target proteins of thioredoxin1 in the social amoeba Dictyostelium discoideum, which possesses three developmentally regulated thioredoxin genes, all of which lack functional characterisation. A two-hybrid approach identified at least seven bona fide thioredoxin partners, including oxidoreductases, proteins of the ribosomal translation apparatus, and the cytoskeletal protein filopodin. With the exception of ribonucleotide reductase, none of these systems had previously been linked to specific redox modulation. Molecular interactions in two of the new thioredoxin/target protein couples were verified by biochemical studies: (1) thioredoxin1 and the abundant elongation factor 1α from D. discoideum form the mixed heterodisulfide characteristic of the thioredoxin mechanism of action; and (2) reduced thioredoxin, but not glutathione, strongly inhibits yeast alcohol dehydrogenase catalysis of ethanol oxidation. ©2004 by Walter de Gruyter Berlin New York Biochemistry nationallicence Molecular biology nationallicence Cellular biology nationallicence alcohol dehydrogenase nationallicence disulfide formation nationallicence elongation factors nationallicence enzyme regulation nationallicence thioredoxins nationallicence thioredoxin oligomers nationallicence Brodegger Thomas Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut Stockmann Anja Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut Oberstraß Jürgen Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut Nellen Wolfgang Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut Follmann Hartmut Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut Biological Chemistry Walter de Gruyter 385/12(2004-12-01), 1185-1192 1431-6730 385:12<1185 2004 385 bchm https://doi.org/10.1515/BC.2004.153 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/BC.2004.153 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Brodegger Thomas Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut NATIONALLICENCE 700 1- Stockmann Anja Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut NATIONALLICENCE 700 1- Oberstraß Jürgen Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut NATIONALLICENCE 700 1- Nellen Wolfgang Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut NATIONALLICENCE 700 1- Follmann Hartmut Departments of Biochemistry and Genetics, Natural Sciences Faculty, University of Kassel, D-34109 Kassel, Germany aut NATIONALLICENCE 773 0- Biological Chemistry Walter de Gruyter 385/12(2004-12-01), 1185-1192 1431-6730 385:12<1185 2004 385 bchm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter