Detection of guanine-C8-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine adduct as a single spot on thin-layer chromatography by modification of the 32P-postlabeling method

Jpn J Cancer Res. 1994 Feb;85(2):113-7. doi: 10.1111/j.1349-7006.1994.tb02068.x.

Abstract

N-(Deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (dG-C8-PhIP) has been shown to be a major adduct in DNA of rats given [3H]PhIP. However, when DNA from organs of rats fed PhIP was analyzed by the 32P-postlabeling method under standard and adduct-intensification conditions, four adduct spots were observed, and 3',5'-pdGp-C8-PhIP was detected as a minor, not a major, adduct spot. Since the three other major adduct spots were suspected to be those of adducted di- or oligo-nucleotides, the 32P-labeled samples were further treated with nuclease P1 and phosphodiesterase I and found to yield only a single adduct spot. The material in this adduct spot was confirmed to be 5'-pdG-C8-PhIP. Thus, using this newly modified 32P-postlabeling method, dG-C8-PhIP was detected as a major adduct in DNA of rats given PhIP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Autoradiography / methods
  • Carcinogens / metabolism*
  • Chromatography, Thin Layer
  • DNA / metabolism*
  • Deoxyguanosine / analogs & derivatives*
  • Deoxyguanosine / analysis
  • Deoxyguanosine / chemistry
  • Deoxyguanosine / isolation & purification
  • Imidazoles / analysis*
  • Imidazoles / chemistry
  • Imidazoles / isolation & purification
  • Imidazoles / metabolism*
  • Imidazoles / toxicity
  • Lung / drug effects
  • Male
  • Mutagens / metabolism*
  • Phosphorus Radioisotopes
  • Rats
  • Rats, Inbred F344

Substances

  • Carcinogens
  • Imidazoles
  • Mutagens
  • Phosphorus Radioisotopes
  • N-(deoxyguanosin-8-yl)-2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine
  • DNA
  • 2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine
  • Deoxyguanosine