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Detection of glutathione free radical catalyzed by prostaglandin hydroperoxidase present in intact epidermal cells

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:7035267
; ;
The authors report the application of the electron spin resonance technique to detect free radicals catalyzed by prostaglandin hydroperoxidase in intact cells. Studies were done using skin epithelial cells obtained from hairless mice which possess significant prostaglandin H synthase (PHS) activity. A technique was developed using ram seminal vesicles for magnification of the enzymatic activity based on the ability of some free radicals formed by PHS to oxidize glutathione to a thiyl radical which was trapped by DMPO. Using this approach they detected a GSH-DMPO adduct catalyzed by cellular PHS. The addition of glutathione stimulated the formation of GSH-DMPO adduct which suggests that most of the oxidation of GSH occurred extracellularly. The addition of the peroxidase substrate phenol greatly stimulated the oxidation of /sup 3/H-arachidonic acid to prostaglandins. The addition of glutathione peroxidase completely inhibited the oxidation of arachidonic acid and the formation of the GSH-DMPO radical adduct. These results suggest that prostaglandin H synthase dependent co-oxidation of chemicals can result in the intracellular formation of free radical metabolites.
Research Organization:
National Institute of Environmental Health Sciences, Research Triangle Park, NC
OSTI ID:
7035267
Report Number(s):
CONF-8604222-
Conference Information:
Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:4
Country of Publication:
United States
Language:
English

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Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL TISSUES
ANIMALS
ARACHIDONIC ACID
BIOSYNTHESIS
BODY
CARBOXYLIC ACIDS
DRUGS
ELECTRON SPIN RESONANCE
ENZYME ACTIVITY
ENZYMES
EPITHELIUM
GLUTATHIONE
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
MAGNETIC RESONANCE
MAMMALS
METABOLISM
METABOLITES
MICE
MONOCARBOXYLIC ACIDS
ORGANIC ACIDS
ORGANIC COMPOUNDS
ORGANS
OXIDOREDUCTASES
PEPTIDES
PEROXIDASES
POLYPEPTIDES
PROSTAGLANDINS
PROTEINS
RADICALS
RADIOPROTECTIVE SUBSTANCES
RESONANCE
RODENTS
SKIN
SYNTHESIS
TISSUES
TRACER TECHNIQUES
TRITIUM COMPOUNDS
VERTEBRATES