title: A study of the kinetic mechanism followed by glutathione reductase from mycelium of Phycomyces blakesleeanus.
abstract: An investigation of the reaction mechanism of glutathione reductase isolated from the mycelium of Phycomyces blakesleeanus NRRL 1555(-) was conducted. The enzyme showed GSSG concentration-dependent substrate inhibition by NADPH and pH-dependent substrate inhibition by GSSG. At pH 7.5, the kinetic data were consistent with a basic scheme corresponding to the branching mechanism, involving a ping-pong with formation of a dead-end F.NADPH complex and an ordered sequential mechanism. Both pathways have in common the step in which NADPH binds to the free oxidized form (E) of the glutathione reductase. At low concentrations of GSSG the ping-pong mechanism prevails, whereas at high concentrations the ordered mechanism appears to dominate. The data were analyzed on the basis of the limiting ping-pong mechanism with F.NADPH complex formation and of the hybrid mechanism, and the kinetic constants of the model were calculated. The data obtained at acidic pH values do not rule out the possibility that the kinetic model may be more complicated than the basic scheme studied.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Universidad de León, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Montero
authors.forename: D
authors.initials: D
authors.lastname: de Arriaga
authors.forename: F
authors.initials: F
authors.lastname: Busto
authors.forename: J
authors.initials: J
authors.lastname: Soler
mesh_terms: D005978:Glutathione / Q000031:analogs & derivatives / Q000494:pharmacology; D019803:Glutathione Disulfide; D005980:Glutathione Reductase / Q000378:metabolism*; D007700:Kinetics; D008433:Mathematics; D008962:Models, Theoretical; D009090:Mucorales / Q000201:enzymology*; D009249:NADP / Q000494:pharmacology; D010084:Oxidation-Reduction; D010800:Phycomyces / Q000201:enzymology*
chemical_list: D009249:NADP; D005980:Glutathione Reductase; D005978:Glutathione; D019803:Glutathione Disulfide