title: Purification and Characterization of Glucose 6-Phosphate Dehydrogenase, 6-Phosphogluconate Dehydrogenase, and Glutathione Reductase from Rat Heart and Inhibition Effects of Furosemide, Digoxin, and Dopamine on the Enzymes Activities.
abstract: The present study was aimed to investigate characterization and purification of glucose-6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and glutathione reductase from rat heart and the inhibitory effect of three drugs. The purification of the enzymes was performed using 2',5'-ADP sepharose 4B affinity material. The subunit and the natural molecular weights were analyzed by SDS-PAGE and gel filtration. Biochemical characteristics such as the optimum temperature, pH, stable pH, and salt concentration were examined for each enzyme. Types of product inhibition and Ki values with Km and Vmax values of the substrates and coenzymes were determined. According to the obtained Ki and IC50 values, furosemide, digoxin, and dopamine showed inhibitory effect on the enzyme activities at low millimolar concentrations in vitro conditions. Dopamine inhibited the activity of these enzymes as competitive, whereas furosemide and digoxin inhibited the activity of the enzyme as noncompetitive.
authors.affiliation: Department of Chemistry, Faculty of Science, Cankiri Karatekin University, 18000, Cankiri, Turkey. sevkiadem@gmail.com.
authors.forename: Sevki
authors.initials: S
authors.lastname: Adem
authors.affiliation: Department of Chemistry, Arts and Science Faculty, Bingol University, 12000, Bingol, Turkey.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Ciftci
mesh_terms: D000818:Animals; D001667:Binding, Competitive; D004077:Digoxin / Q000737:chemistry*; D004298:Dopamine / Q000737:chemistry*; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000737:chemistry*; D005665:Furosemide / Q000737:chemistry*; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D005980:Glutathione Reductase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008297:Male; D008970:Molecular Weight; D009206:Myocardium / Q000737:chemistry / Q000201:enzymology; D010734:Phosphogluconate Dehydrogenase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification*; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004791:Enzyme Inhibitors; D004077:Digoxin; D005665:Furosemide; D010734:Phosphogluconate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D005980:Glutathione Reductase; D004298:Dopamine
keywords: 6-Phosphogluconate dehydrogenase; Drug inhibition; Glucose 6-phosphate dehydrogenase; Glutathione reductase; Heart; Protein purification and characterization; Rats (Sprague-Dawley)