sup 32 P-Postlabeling measurement of adenine N-1-oxide in cellular DNA exposed to hydrogen peroxide
- Centre d'Etudes Nucleaires de Grenoble (France)
A {sup 32}P-postlabeling assay has been developed for monitoring the formation within DNA of adenine N-1-oxide, the specific H{sub 2}O{sub 2}-mediated oxidation product under nonradical conditions. This has required the chemical synthesis of both 2{prime}-deoxyadenosine N-1-oxide 3{prime}-monophosphate and 2{prime}-deoxyadenosine N-1-oxide 5{prime}-monophosphate, the substrate and the product of polynucleotide kinase mediated phosphorylation. Isolation of the substrate from the other nucleotides was found to be necessary in order to improve the rate of phosphorylation and to prevent self-radiolysis processes. ({sup 32}P)-2{prime}-deoxyadenosine N-1-oxide 5{prime}-monophosphate was obtained after successive ion exchange and reverse-phase HPLC and was characterized by a microreaction. The sensitivity of the assay, which is close to 1 modified adenine N-1-oxide/10{sup 6} bases, allowed the determination of this lesion within the DNA of cells exposed to nonlethal levels of H{sub 2}O{sub 2}.
- OSTI ID:
- 6411490
- Journal Information:
- Chemical Research in Toxicology; (USA), Vol. 3:2; ISSN 0893-228X
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
DNA
OXIDATION
ADENINES
HYDROGEN PEROXIDE
ION EXCHANGE
LIQUID COLUMN CHROMATOGRAPHY
PHOSPHORUS 32
AMINES
ANTIMETABOLITES
AROMATICS
AZAARENES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMICAL REACTIONS
CHROMATOGRAPHY
DAYS LIVING RADIOISOTOPES
DRUGS
HETEROCYCLIC COMPOUNDS
HYDROGEN COMPOUNDS
ISOTOPES
LIGHT NUCLEI
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
OXYGEN COMPOUNDS
PEROXIDES
PHOSPHORUS ISOTOPES
PURINES
RADIOISOTOPES
SEPARATION PROCESSES
560300* - Chemicals Metabolism & Toxicology