Radiochemical high-performance liquid chromatographic assay for the determination of catechol O-methyltransferase activity towards various substrates
作者： Pia Lautala , Ismo Ulmanen , Jyrki Taskinen
DOI: 10.1016/S0378-4347(99)00455-7
关键词:
摘要: A new chromatographic catechol O-methyltransferase (COMT) assay based on S-adenosyl-L-[methyl-14C]methionine and on-line radioactivity detection was developed. With minor modifications in the mobile phase composition methylation velocities for 30 structurally diverse compounds including simple catechols, neurotransmitters, catecholestrogens catecholic drugs could be measured using human rat recombinant soluble COMT. The enzymes showed very similar substrate selectivities. radiochemical method validated 3,4-dihydroxybenzoic acid as a model it shown that accurate reproducible velocity values achieved both of hydroxyls. proved to suited determining enzyme kinetic parameters can probably further used gathering data differentially substituted catechols order construct proper structure-activity relationships
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