Molecular Cloning, Expression, purification and Characterization of the Zebrafish Catechol-O-methyltransferases

The zebrafish is emerging as an important animal model for biomedical research. This study represents part of an effort to establish the zebrafish as a model for investigating the methylation pathway of drug metabolism. Two zebrafish catechol O-methyltransferases (COMTs), designated COMT-1 and COMT-2, were cloned, expressed, purified, and characterized. Substrate specificity of these two zebrafish COMTs were analyzed using a panel of catechol drugs including dobutamine, methyldopa, and isoproterenol, as well as endogenous catecholestrogens, catecholamines, and dopa. The expression of COMT-1 was examined from embryogenesis to maturity by employing both the conventional RT-PCR and RT-quantitative PCR. Data on the COMT-1 expression provided insights into the developmental expression of the COMT enzyme, which may serve as a basis for further investigation into its possible protective role against the adverse effects of catechol drugs and other xenobiotic catechols during the developmental process.