The mechanism of CeCl3 on the activiation of alanine aminotransferase from mice.

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 CeCl(3) and ALT. In this report, we investigated the mechanisms of CeCl(3) on ALT activity in vivo and in vitro. Our results showed that Ce(3+) could significantly activate ALT in vivo and in vitro; the kinetics constant (Km) and Vmax were 0.018 microM and 1,380 unit mg(-1) protein min(-1), respectively, at a low concentration of Ce(3+), and 0.027 microM and 624 unit mg(-1) protein min(-1), respectively, at a high concentration of Ce(3+). By UV absorption and fluorescence spectroscopy assays, the Ce(3+) was determined to be directly bound to ALT; the binding site of Ce(3+) to ALT was 1.72, and the binding constants of the binding site were 4.82 x 10(8) and 9.05 x 10(7) L mol(-1). Based on the analysis of the circular dichroism spectra, it was concluded that the binding of Ce(3+) 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 Ce(3+).