/sup 32/P-postlabeling analysis of non-radioactive aromatic carcinogen--DNA adducts

Journal Article · Fri Jan 01 04:00:00 EST 1982 · Carcinogenesis (N.Y.); (United States)

DOI:https://doi.org/10.1093/carcin/3.9.1081· OSTI ID:6521583

Gupta, R C; Reddy, M V; Randerath, K

A newly developed enzymatic /sup 32/P-postlabeling method was applied to the analysis of DNA's containing non-radioactive arylamine, arylamide, and polycyclic aromatic hydrocarbon adducts. DNA reacted in vitro with N-hydroxy-2-amino-fluorene, N-acetoxy-2-acetylaminofluorene, and 7 beta,8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene, respectively, as well as DNA preparations from the liver of rats treated with N-hydroxy-2-acetylaminofluorene and benzo(a)pyrene, respectively, were enzymatically digested to deoxyribonucleoside 3'-monophosphates, which were then converted to (5'-/sup 32/P)deoxyribonucleoside 3',5'-bisphosphates by T4 polynucleotide kinase-catalyzed (/sup 32/P)phosphate transfer from (gamma-/sup 32/P)ATP. The /sup 32/P-labeled nucleotides were resolved by anion-exchange t.l.c. on polyethyleneimine-cellulose and detected by autoradiography. Aromatic adduct nucleotides were found to be retained at the origin in aqueous electrolyte solutions, but to migrate as distinct spots in solvents containing 7-8.5 M urea. Advantage was taken of this observation to remove /sup 32/P-labeled normal DNA nucleotides from adduct nucleotides. This purification enabled the detection of a single adduct in 10(7)-10(8) normal nucleotides. The method appears applicable to the ultrasensitive detection of a large number of carcinogen--DNA adducts of diverse structure without requiring radioactive carcinogens or specific antibodies.

• Research Organization: Department of Pharmacology, Baylor College of Medicine, Texas Medical Center, Houston
• OSTI ID: 6521583
• Journal Information: Carcinogenesis (N.Y.); (United States), Journal Name: Carcinogenesis (N.Y.); (United States) Vol. 3:9; ISSN CRNGD
• Country of Publication: United States
• Language: English

Related Subjects

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560305* -- Chemicals Metabolism & Toxicology-- Vertebrates-- (-1987)
62 RADIOLOGY AND NUCLEAR MEDICINE
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
AROMATICS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOCHEMISTRY
BIOLOGICAL EFFECTS
BIOLOGICAL PATHWAYS
CARCINOGENESIS
CARCINOGENS
CHEMICAL PREPARATION
CHEMISTRY
DAYS LIVING RADIOISOTOPES
DNA
HYDROCARBONS
ISOTOPE APPLICATIONS
ISOTOPES
LABELLING
LIGHT NUCLEI
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PATHOGENESIS
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
POLYCYCLIC AROMATIC HYDROCARBONS
RADIOISOTOPES
SYNTHESIS