Radiometric assay for phenylethanolamine N-methyltransferase and catechol O-methyltransferase in a single tissue sample: application to rat hypothalamic nuclei, pineal gland, and heart
Journal Article
·
· Anal. Biochem.; (United States)
A simple and highly sensitive method for simultaneous assay of phenylethanolamine N-methyltransferase (PNMT) and catechol O-methyltransferase (COMT) is described. These enzymes are determined in a single tissue homogenate using S-(methyl-/sup 3/H) adenosyl-L-methionine as methyl donor and sequentially incubating with the substrates phenylethanolamine and epinephrine. The radioactive products of the enzymatic reactions, N-methylphenylethanolamine and metanephrine, are extracted and then separated by thin-layer chromatography. The identity of the reaction products has been established chromatographically and the conditions for both enzymatic reactions in the assay procedure have been defined. Measurement of PNMT activity in the rat pineal gland or in minute fragments of other tissues (e.g., brain nuclei) has not been possible using previously described methods. Activities of PNMT and COMT in the rat pineal gland, various hypothalamic nuclei, and the auricular and ventricular myocardia are herein reported.
- Research Organization:
- Institute of Experimental Endocrinology, Bratislava, Czechoslovakia
- OSTI ID:
- 5856851
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 164:2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
BODY
BRAIN
CARBOXYLIC ACIDS
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
CELL NUCLEI
CENTRAL NERVOUS SYSTEM
CHROMATOGRAPHY
DRUGS
ENZYME ACTIVITY
ENZYMES
GLANDS
HEART
HYPOTHALAMUS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPOTROPIC FACTORS
MAMMALS
METHIONINE
MUSCLES
MYOCARDIUM
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PINEAL GLAND
RADIOASSAY
RATS
RODENTS
SEPARATION PROCESSES
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES
59 BASIC BIOLOGICAL SCIENCES
AMINO ACIDS
ANIMALS
BODY
BRAIN
CARBOXYLIC ACIDS
CARDIOVASCULAR SYSTEM
CELL CONSTITUENTS
CELL NUCLEI
CENTRAL NERVOUS SYSTEM
CHROMATOGRAPHY
DRUGS
ENZYME ACTIVITY
ENZYMES
GLANDS
HEART
HYPOTHALAMUS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIPOTROPIC FACTORS
MAMMALS
METHIONINE
MUSCLES
MYOCARDIUM
NERVOUS SYSTEM
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANIC SULFUR COMPOUNDS
ORGANS
PINEAL GLAND
RADIOASSAY
RATS
RODENTS
SEPARATION PROCESSES
THIN-LAYER CHROMATOGRAPHY
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES