Vol. 301, Issue 1, 315-321, April 2002

Kinetic Interactions of Dopamine and Dobutamine with Human Catechol-O-methyltransferase and Monoamine Oxidase in Vitro

Maohe Yan, Leslie T. Webster, Jr. and Jeffrey L. Blumer

Departments of Pediatrics and Pharmacology, Case Western Reserve University, Division of Pediatric Pharmacology and Critical Care, Rainbow Babies and Children's Hospital of the University Hospitals of Cleveland, Cleveland, Ohio

Dopamine and dobutamine are often infused together into acutely ill patients requiring temporary support of cardiac and renal function, but whether these catecholamines affect the metabolic clearance of each other is not established. We determined the kinetics of dopamine and dobutamine as substrates and inhibitors of each other, i.e., apparent V_max, K_m, and K_i, with crude preparations of human blood mononuclear cell catechol-O-methyltransferase (COMT) and platelet monoamine oxidase (MAO) at pH 7.4 and 37°C. Values of V_max for dopamine and dobutamine as substrates for COMT were 0.45 and 0.59 nmol of 3-O-methyl product formed per milligram of protein per minute, whereas those for K_m were 0.44 and 0.05 mM, respectively. Dopamine and dobutamine were competitive inhibitors of each other in this reaction. The K_i for dopamine as an inhibitor of dobutamine methylation was 1.5 mM, whereas that for dobutamine as an inhibitor of dopamine methylation was 0.015 mM. Dopamine but not dobutamine was a substrate for MAO. The V_max for dihydroxyphenylacetaldehyde formation from dopamine was 0.29 nmol/mg protein/min and the K_m for dopamine was 0.38 mM. Dobutamine was a noncompetitive inhibitor of dopamine oxidation in this reaction (K_i  1.19 mM). The high apparent K_m and K_i values derived for dopamine and dobutamine when tested with these two human enzymes in vitro suggest that these catecholamines do not interfere with the metabolism of each other when both are infused together at therapeutic concentrations.