New spectrophotometric and radiochemical assays for acetyl-CoA: arylamine N-acetyltransferase applicable to a variety of arylamines
Journal Article
·
· Anal. Biochem.; (United States)
Simple and sensitive spectrophotometric and radiochemical procedures are described for the assay of acetyl-CoA:arylamine N-acetyltransferase (NAT), which catalyzes the reaction acetyl-CoA + arylamine----N-acetylated arylamine + CoASH. The methods are applicable to crude tissue homogenates and blood lysates. The spectrophotometric assay is characterized by two features: (i) NAT activity is measured by quantifying the disappearance of the arylamine substrate as reflected by decreasing Schiff's base formation with dimethylaminobenzaldehyde. (ii) During the enzymatic reaction, the inhibitory product CoASH is recycled by the system acetyl phosphate/phosphotransacetylase to the substrate acetyl-CoA. The radiochemical procedure depends on enzymatic synthesis of (/sup 3/H)acetyl-CoA in the assay using (/sup 3/H)acetate, ATP, CoASH, and acetyl-CoA synthetase. NAT activity is measured by quantifying N-(/sup 3/H)acetylarylamine after separation from (/sup 3/H)acetate by extraction. Product inhibition by CoASH is prevented in this system by the use of acetyl-CoA synthetase.
- Research Organization:
- Univ. of Michigan Medical School, Ann Arbor
- OSTI ID:
- 5253617
- Journal Information:
- Anal. Biochem.; (United States), Journal Name: Anal. Biochem.; (United States) Vol. 2; ISSN ANBCA
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ACETATES
AMINES
ANIMALS
BIOASSAY
BODY
CARBOXYLIC ACID SALTS
CATALYSIS
CHEMISTRY
DIGESTIVE SYSTEM
ENZYME ACTIVITY
ENZYMES
GLANDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIGASES
LIVER
MAMMALS
MICE
ORGANIC COMPOUNDS
ORGANS
RABBITS
RADIOCHEMISTRY
RODENTS
SPECTROPHOTOMETRY
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES
59 BASIC BIOLOGICAL SCIENCES
ACETATES
AMINES
ANIMALS
BIOASSAY
BODY
CARBOXYLIC ACID SALTS
CATALYSIS
CHEMISTRY
DIGESTIVE SYSTEM
ENZYME ACTIVITY
ENZYMES
GLANDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIGASES
LIVER
MAMMALS
MICE
ORGANIC COMPOUNDS
ORGANS
RABBITS
RADIOCHEMISTRY
RODENTS
SPECTROPHOTOMETRY
TRACER TECHNIQUES
TRANSFERASES
TRITIUM COMPOUNDS
VERTEBRATES