Analysis of binding interaction between puerarin and bovine serum albumin by multi-spectroscopic method.

The interaction of puerarin and bovine serum albumin (BSA) was investigated by means of fluorescence spectroscopy, resonance light-scattering spectroscopy, infrared spectroscopy, and synchronous fluorescence spectra. The apparent binding constants (K(a)) between puerarin and BSA were 1.13 x 10(4) (20 degrees C), and 1.54 x 10(4) lmol(-1) (30 degrees C), and the binding sites values (n) were 0.95+/-0.02. The experimental results showed that the puerarin could be inserted into the BSA, quenching the inner fluorescence by forming the puerarin-BSA complex. The addition of increasing puerarin to BSA solution leads to the gradual decrease in RLS intensity, exhibiting the formation of the aggregate in solution. It was found that both static quenching and non-radiation energy transfer were the main reasons for the fluorescence quenching. The positive entropy change and enthalpy change indicated that the interaction of puerarin and BSA was driven mainly by hydrophobic forces. The process of binding was a spontaneous process in which Gibbs free energy change was negative. The competing binding reaction with BSA between Fe(3+), Cu(2+) and puerarin was investigated. The effect of Fe(3+) and Cu(2+) on the binding of puerarin with BSA is discussed.