Kinetic and equilibrium studies on the interaction of reduced flavoprotein D-amino acid oxidase with pyridine carboxylates.
The equilibrium constants and the rate constants (binding and dissociation constants) between reduced D-amino acid oxidase and pyridine carboxylates were obtained at various pH values (from pH 6.0 to 8.3). The pH dependence of the constants is consistent with the previous conclusion from a resonance Raman study that pyridine carboxylates in the form of a cation protonated at the N atom can bind to the reduced enzyme, but those in the neutral form cannot bind, showing that the positive charge of cationic pyridine carboxylates interacts with the negative charge of the anionic reduced flavin in the reduced enzyme. The binding rate constants of picolinate and nicotinate in the cationic form for the reduced enzyme were quite similar to each other, but the dissociation rate constant of picolinate is several times smaller than that of nicotinate. Thus, it is concluded that the difference in affinity of picolinate and nicotinate for the reduced enzyme is derived from the difference of the dissociation rate constants.
D000818:Animals; D003605:D-Amino-Acid Oxidase / Q000378:metabolism*; D005420:Flavoproteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007668:Kidney / Q000201:enzymology; D007700:Kinetics; D009525:Niacin / Q000378:metabolism*; D010084:Oxidation-Reduction; D010848:Picolinic Acids / Q000378:metabolism*; D011485:Protein Binding; D013552:Swine
D005420:Flavoproteins; D010848:Picolinic Acids; D009525:Niacin; D003605:D-Amino-Acid Oxidase; C030614:picolinic acid