Effects of different valences of cerium ion on conformation of Horseradish Peroxidase

Our previous studies demonstrated that Ce4+ could induce reactive oxygen species (ROS) burst as a signal to promote pacilitaxel biosynthesis in suspension cultured Taxus cuspidate cells. To further understand the mechanism of cerium ions inducing ROS burst, circular dichroism (CD), synchronous fluorescence, and electron paramagnetic resonance (EPR) were used to detect them inducing conformational change of horseradish peroxidase (HRP). Horseradish peroxidase activity was reduced by 78% by 0.1 mmol/L Ce4+, whereas it was only reduced by 28% by 0.1 mmol/L Ce3+. Circular dichroism spectra showed that the percentage of transition from helical content and other structure to β strands and β turns was 23.1 when induced by Ce4+, whereas it was only 13.2 when induced by Ce3+. In synchronous fluorescence spectra, Ce4+ led to red shift and intensity-elevation of tryptophan fluorescence emission maximum, whereas in the case of Ce3+, the results were a contrast to the above. Furthermore, g factor (gx and gy) in electron paramagnetic resonance (EPR) induced by Ce4+ and Ce3+ was significantly different. These results indicated that the different valence of cerium ion induced various conformations of HRP, and Ce4+ was more effective than Ce3+. This suggested that Ce4+ affected the burst of ROS through changing the conformation of oxidoreductase.