title: Dimer-tetramer equilibrium of glutathione reductase from the cyanobacterium Spirulina maxima.
abstract: Glutathione reductase [NAD(P)H:GSSG oxidoreductase; EC 1.6.4.2] from cyanobacterium Spirulina maxima exists as an equilibrium system between a dimer (S20,W = 5.96) and a tetramer (S20,W = 8.49) which has a very slow interconversion rate at neutral pH. Our results showed that the apparent dissociation constant (kd) was 4.61 X 10(-7) M. The proportion of both forms at pH 7.0 did not alter at either 4 or 25 degrees C. However, electrophoretic analysis at various pH values showed that at 25 degrees C a gradual transition takes place between oligomers with an apparent pKa of 7.55. When dimers aggregate to form tetramers, the reaction involves the uptake of eight protons (K = 1.58 X 10(-64) M9). At pH 7.7, the equilibrium shifts completely from dimers-tetramers to dimers when temperature is increased, which would suggest that the dissociation is an endothermic process. Thermodynamic parameters obtained from the temperature study show that the dissociation of glutathione reductase is characterized by positive entropy and enthalpy changes. Neither NADPH nor GSSG have any effect on the dimer-tetramer equilibrium. Measurements of reductase activity indicate that the tetramer is almost certainly active, whereas the dimer is either less active or inactive.
authors.affiliation: Departamento de Bioquímica, Facultad de Medicina, Universidad Nacional Autónoma de México, Mexico City, D.F.
authors.forename: J L
authors.initials: JL
authors.lastname: Rendón
authors.forename: G
authors.initials: G
authors.lastname: Mendoza-Hernández
mesh_terms: D000458:Cyanobacteria / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D005980:Glutathione Reductase / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D013816:Thermodynamics; D014461:Ultracentrifugation / Q000379:methods
chemical_list: D046911:Macromolecular Substances; D005980:Glutathione Reductase