Determination of catechol-O-methyltransferase activity in brain tissue by high-performance liquid chromatography with on-line radiochemical detection
A sensitive assay for catechol-O-methyltransferase (COMT) activity by high-performance liquid chromatography with on-line radiochemical detection was described. The method was based on the measurement of /sup 3/H- labeled 3-O- and 4-O-methylated products of the substrate, 3,4- dihydroxybenzoic acid, using S-adenosyl-L-(methyl-/sup 3/H)methionine as the methyl donor, or the measurement of /sup 14/C-labeled 3-O- and 4-O-methylated products of the substrate, (7-/sup 14/C)dopamine. The reaction products were determined from the incubation mixture after removal of protein by injecting an aliquot into the liquid chromatograph. The detection limit with counting efficiency of 40% was 0.45 pmol /sup 3/H-labeled product, and 0. 04 pmol /sup 14/C-labeled product with 61% counting efficiency. The method is suitable for assaying membrane-bound and soluble COMT activities in the brain tissue and for calculation of meta/para product ratios.
- Research Organization:
- Orion Pharmaceutical Co. Research Center, Espoo, Finland
- OSTI ID:
- 5554387
- Journal Information:
- Anal. Biochem.; (United States), Vol. 144:1
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
TRANSFERASES
ENZYME ACTIVITY
BENZOIC ACID
BIOASSAY
BIOCHEMICAL REACTION KINETICS
BRAIN
CARBON 14 COMPOUNDS
DOPAMINE
LIQUID COLUMN CHROMATOGRAPHY
METHIONINE
SUBSTRATES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
AMINES
AMINO ACIDS
AROMATICS
AUTONOMIC NERVOUS SYSTEM AGENTS
BODY
CARBOXYLIC ACIDS
CARDIOTONICS
CARDIOVASCULAR AGENTS
CENTRAL NERVOUS SYSTEM
CHROMATOGRAPHY
DRUGS
ENZYMES
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
KINETICS
LABELLED COMPOUNDS
LIPOTROPIC FACTORS
MONOCARBOXYLIC ACIDS
NERVOUS SYSTEM
NEUROREGULATORS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PHENOLS
POLYPHENOLS
REACTION KINETICS
SEPARATION PROCESSES
SYMPATHOMIMETICS
550201* - Biochemistry- Tracer Techniques