title: Purification and kinetic properties of 6-phosphogluconate dehydrogenase from rat small intestine.
abstract: 6-Phosphogluconate dehydrogenase (6PG) was purified from rat small intestine with 36% yield and a specific activity of 15 U/mg. On SDS/PAGE, one band with a mass of 52 kDa was found. On native PAGE three protein and two activity bands were observed. The pH optimum was 7.35. Using Arrhenius plots, Ea, DeltaH, Q10 and Tm for 6PGD were found to be 7.52 kcal/mol, 6.90 kcal/mol, 1.49 and 49.4 degrees C, respectively. The enzyme obeyed "Rapid Equilibrium Random Bi Bi" kinetic model with Km values of 595 +/- 213 microM for 6PG and 53.03+/-1.99 microM for NADP. 1/Vm versus 1/6PG and 1/NADP plots gave a Vm value of 8.91+/-1.92 U/mg protein. NADPH is the competitive inhibitor with a Ki of 31.91+/-1.31 microM. The relatively small Ki for the 6PGD:NADPH complex indicates the importance of NADPH in the regulation of the pentose phosphate pathway through G6PD and 6PGD.
authors.affiliation: Department of Biochemistrry, Faculty of Medicine, Hacettepe University, 06100, Ankara, Turkey.
authors.forename: Deniz
authors.initials: D
authors.lastname: Ceyhan
authors.forename: Ali
authors.initials: A
authors.lastname: Danişan
authors.forename: I Hamdi
authors.initials: IH
authors.lastname: Oğüş
authors.forename: Nazmi
authors.initials: N
authors.lastname: Ozer
mesh_terms: D000818:Animals; D004789:Enzyme Activation; D005260:Female; D006863:Hydrogen-Ion Concentration; D007421:Intestine, Small / Q000201:enzymology*; D007700:Kinetics; D008970:Molecular Weight; D010734:Phosphogluconate Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D013696:Temperature; D013816:Thermodynamics
chemical_list: D010734:Phosphogluconate Dehydrogenase