Chromium (Cr) poses potential threats on human and ecological health through both food and environmental pathways. However, the response mechanisms of Cr to function-related proteins in human subjects and the differential effects in the case of different valence states of Cr are still lacking. Herein, the binding interactions and mechanisms of lysozyme to Cr(III) and Cr(VI) were investigated by isothermal titration microcalorimetry, multi-spectroscopy, enzyme activity assay, and molecular simulations. Our results indicated that the binding of Cr in different valence states with lysozyme was a relatively affinity process and mainly driven by hydrophobic forces. Both Cr species induced the looseness of backbone for lysozyme and the enhancement of hydrophilicity around the backbone microenvironment. Otherwise, the alpha-helix of lysozyme was decreased with different valence states of Cr, while Cr(III) showed a greater degree of influence and a stronger regularity. Interestingly, no significant changes in the microenvironment around aromatic amino acid residues were observed under the exposure of different valence states of Cr. Differential structural changes of lysozyme resulted in activity alter-ations. At 80 mg center dot L-1 of Cr, Cr (VI) reduced the lysozyme activity to 81.44% of the control, while the lysozyme activity was significantly reduced to 24.45% under the exposure of Cr (III). Further molecular docking showed that amino acids near the binding sites of different valence states of Cr with lysozyme were not identical, but the enzyme active site, Asp52, was all within the binding pocket. This study elucidates the differential toxicity effects and interaction mechanisms of Cr in different valence states, and offers a comprehensive strategy for exploring the in vitro responses of toxicants exposure.
