Interaction of ferredoxin-NADP+ reductase from Anabaena with its substrates.
The interaction of ferredoxin-NADP+ reductase from the cyanobacterium Anabaena variabilis with its substrates, NADP+ and ferredoxin, has been studied by difference absorption spectroscopy. Several structural analogs of NADP+ have been shown to form complexes the stabilities of which are strongly dependent on the ionic strength of the medium. In most cases the binding energy of these complexes and their difference absorption spectra are similar to those reported for the spinach enzyme. However, NADP+ perturbs the absorption spectra of the Anabaena and spinach enzymes in a different way. This difference has been shown to be related to the binding of the nicotinamide ring of NADP+ to the enzymes. These results are interpreted as being due to a different nicotinamide binding site in the two reductases. The enthalpic and entropic components of the Gibbs energy of formation of the NADP+ complex have been estimated. An increase in entropy on NADP+ binding seems to be the main source of stability for the complex. A shift of approximately 40 mV in the redox potential of the couple NADP+/NADPH has been observed to occur upon binding of NADP+ to the oxidized enzyme. This allows us to calculate the binding energy between the reductase and NADPH. The ability of the reductase, ferredoxin, and NADP+ to form a ternary complex indicates that the protein carrier binds to the reductase through a different site than that of the pyridine nucleotide.
Sancho
J
J
Departamento de Bioquímica y Biología Molecular y Cellular, Facultad de Ciencias, Universidad de Zaragoza, Spain.
Gómez-Moreno
C
C
D001665:Binding Sites; D000458:Cyanobacteria / Q000201:enzymology*; D004579:Electron Transport; D005287:Ferredoxin-NADP Reductase / Q000378:metabolism*; D005288:Ferredoxins; D005418:Flavodoxin; D009249:NADP; D010084:Oxidation-Reduction; D013053:Spectrophotometry; D013379:Substrate Specificity; D013816:Thermodynamics
D005288:Ferredoxins; D005418:Flavodoxin; D009249:NADP; D005287:Ferredoxin-NADP Reductase