Assessment of catechol-O-methyltransferase activity and its inhibition in erythrocytes of animals and humans.
A non-radiometric method to measure the catechol-O-methyltransferase (COMT) activity in erythrocytes was modified to increase its sensitivity four-fold as well as its reproducibility and applicability. The method is based on the COMT-mediated O-methylation of 4-(naphtho [1,2-d] thiazol-2-yl) pyrocatechol, the product of which was determined fluorometrically. COMT activities down to less than 1% of those present at baseline could be measured precisely and accurately. The intra- and inter-assay coefficients were below 3 and 5.3%, respectively. Basal COMT activity and the distribution between soluble and membrane-bound COMT was shown to be variable among different species (ten species tested). The applicability of the method was demonstrated by the characterization of COMT activity-time courses in human erythrocytes after oral administration of the COMT inhibitor tolcapone. The assay developed will be useful in the rapid screening and clinical development of new COMT inhibitors.[1]References
- Assessment of catechol-O-methyltransferase activity and its inhibition in erythrocytes of animals and humans. Zürcher, G., Da Prada, M., Dingemanse, J. Biomed. Chromatogr. (1996) [Pubmed]
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