title: Purification and biochemical characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase from rat lung and inhibition effects of some antibiotics.
abstract: G6PD, 6PGD and GR have been purified separately in the single step from rat lung using 2', 5'-ADP Sepharose 4B affinity chromatography. The purified enzymes showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of the enzymes were estimated to be 134 kDa for G6PD, 107 kDa for 6PGD and 121 kDa for GR by Sephadex G-150 gel filtration chromatography, and the subunit molecular weights was respectively found to be 66, 52 and 63 kDa by SDS-PAGE. Optimum pH, stable pH, optimum ionic strength, optimum temperature, KM and Vmax values for substrates were determined. Product inhibition studies were also performed. The enzymes were inhibited by levofloxacin, furosemide, ceftazidime, cefuroxime and gentamicin as in vitro with IC50 values in the range of 0.07-30.13 mM. In vivo studies demonstrated that lung GR was inhibited by furosemide and lung 6PGD was inhibited by levofloxacin.
authors.affiliation: a Department of Chemistry , Faculty of Science, Cankiri Karatekin University , Cankiri , Turkey and.
authors.forename: Sevki
authors.initials: S
authors.lastname: Adem
authors.affiliation: b Department of Chemistry , Faculty of Art and Science, Bingol University , Bingol , Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Ciftci
mesh_terms: D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D005980:Glutathione Reductase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D008168:Lung / Q000201:enzymology*; D010734:Phosphogluconate Dehydrogenase / Q000037:antagonists & inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000900:Anti-Bacterial Agents; D004791:Enzyme Inhibitors; D010734:Phosphogluconate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D005980:Glutathione Reductase
keywords: 6-phosphogluconate dehydrogenase (6PGD); Drug inhibition; glucose 6-phosphate dehydrogenase (G6PD); glutathione reductase (GR); lung; protein purification and characterization; rats (Sprague–Dawley)