title: Purification and partial characterization of thioredoxin reductase from Streptomyces aureofaciens.
abstract: Thioredoxin reductase (TrxR) is one of a number of flavoproteins that catalyze the transfer of electrons between pyridine nucleotides and a specific disulfide-containing substrate. Thioredoxin reductase from Streptomyces aureofaciens 3239 has been purified to homogeneity by a two-step chromatographic procedure including anion-exchange chromatography and affinity chromatography on 2'5'-ADP-Sepharose 4B. Molar mass determined by chromatography on Superose 12 HR 10/30 and sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed 69 kDa for the native protein and 34.8 kDa for the enzyme subunit. The isoelectric point determined by isoelectric focusing gel electrophoresis was 4.3. TrxR effectively catalyzed the reduction of DTNB in the presence of S. aureofaciens thioredoxin-1. TrxR activity in the presence of S. aureofaciens thioredoxin-2 was only 1/4 of the activity with thioredoxin-1 (1). The activity of pure TrxR decreased drastically in the presence of NADPH, while NADP+ as well as Streptomyces aureofaciens thioredoxin-1 protected the enzyme from inactivation. These results indicate that thioredoxin reductase activity in bacteria could be modulated by the redox status of NADP+/NADPH and thioredoxin pools.
authors.affiliation: Department of Biochemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovak Republic.
authors.forename: T
authors.initials: T
authors.lastname: Horecká
authors.forename: D
authors.initials: D
authors.lastname: Perecko
authors.forename: E
authors.initials: E
authors.lastname: Kutejová
authors.forename: D
authors.initials: D
authors.lastname: Mikulásová
authors.forename: M
authors.initials: M
authors.lastname: Kollárová
mesh_terms: D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D008970:Molecular Weight; D013304:Streptomyces aureofaciens / Q000201:enzymology*; D013880:Thioredoxin-Disulfide Reductase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
chemical_list: D013880:Thioredoxin-Disulfide Reductase