The Mechanism of CeCl₃ on the Activiation of Alanine Aminotransferase from Mice

Abstract

The activity of alanine aminotransferase (ALT; E.C. 2.6.1.2) is often changed upon inflammatory responses in animals. Rare earths was shown to provoke various inflammatory responses both in rats and mice; however, the molecular mechanism by which rare earths exert its toxicity has not been completely understood, especially, we know little about the mechanism of the interaction between CeCl₃ and ALT. In this report, we investigated the mechanisms of CeCl₃ on ALT activity in vivo and in vitro. Our results showed that Ce³⁺ could significantly activate ALT in vivo and in vitro; the kinetics constant (Km) and Vmax were 0.018 µM and 1,380 unit mg⁻¹ protein min⁻¹, respectively, at a low concentration of Ce³⁺, and 0.027 µM and 624 unit mg⁻¹ protein min⁻¹, respectively, at a high concentration of Ce³⁺. By UV absorption and fluorescence spectroscopy assays, the Ce³⁺ was determined to be directly bound to ALT; the binding site of Ce³⁺ to ALT was 1.72, and the binding constants of the binding site were 4.82 × 10⁸ and 9.05 × 10⁷ L mol⁻¹. Based on the analysis of the circular dichroism spectra, it was concluded that the binding of Ce³⁺ altered the secondary structure of ALT, suggesting that the observed enhancement of ALT activity was caused by a subtle structural change in the active site through the formation of the complex with Ce³⁺.