Study on the interaction between pyridoxal and CopC by multi-spectroscopy and docking methods.

Abstract The interaction between pyridoxal hydrochloride (HQ) and apoCopC was investigated using Fourier transform infrared spectroscopy (FTIR), isothermal titration calorimetry (ITC), circular dichroism (CD), fluorescence spectroscopy, three-dimensional (3D) fluorescence spectroscopy, fluorescence lifetime, TNS fluorescence and docking methods. FTIR, CD, TNS fluorescence and fluorescence lifetime experiments suggested that the apoCopC conformation was altered by HQ with an increase in the random coil content and a reduction in the β-sheet content. In addition, the data from fluorescence spectroscopy, 3D fluorescence spectroscopy and molecular docking revealed that the binding site of HQ was located in the hydrophobic area of apoCopC, and a redshift of the HQ fluorescence spectra was observed. Furthermore, ITC and fluorescence quenching data manifested that the binding ratio of HQ and apoCopC was 1:1, and the forming constant was calculated to be (7.06 ± 0.21) × 105 M−1. The thermodynamic parameters Δ H and Δ S suggested that the formation of a CopC-HQ complex depended on the hydrophobic force. Furthermore, the average binding distance between tryptophan in apoCopC and HQ was determined by means of Förster non-radioactive resonance energy transfer and molecular docking. The results agreed well with each other. As a redox switch in the modulation of copper, the interaction of apoCopC with small molecules will affect the action of the redox switch. These findings could provide useful information to illustrate the copper regulation mechanism. Graphical Abstract The interaction between pyridoxal hydrochloride and apoCopC has been investigated in details by means of FTIR, CD, ITC, multi-spectroscopy and docking methods. Unlabelled Image Highlights • 1:1 complex formed from HQ and CopC was investigated. • The binding of HQ can result in the change of conformation of CopC. • Red shift of HQ fluorescence spectra was observed. • The distance between the unique Trp in CopC and HQ was measured. [ABSTRACT FROM AUTHOR]