Separation of (+)-syn- and (-)-anti-benzo[a]pyrene dihydrodiol epoxide-DNA adducts in 32P-postlabeling analysis.
The (+)-enantiomer of 7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene (BP-7,8-diol) is a diagnostic probe for cytochrome P-450 and non-cytochrome P-450 pathways of dihydrodiol epoxidation. The principle products of epoxidation are the (+)-syn-dihydrodiol epoxide [(+)-syn-BPDE] and the (-)-anti-dihydrodiol epoxide [(-)-anti-BPDE]. Chromatographic conditions are described that separate the major deoxynucleoside 3',5'-bisphosphate adducts derived from these dihydrodiol epoxides on commercial poly(ethylenimine) thin-layer plates. Inclusion of boric acid and magnesium chloride in the D4 solvent is a key feature of the separation. Reasonable separation of these bisphosphate adducts from the major deoxynucleoside 3',5'-bisphosphate adduct derived from (+)-anti-BPDE is also observed. 32P-Postlabeling analysis of DNA adducts produced following topical administration of benzo[a]pyrene to mouse skin suggests that cytochrome P-450 plays a major role in its metabolism to DNA binding derivatives.
Reddy
A P
AP
Department of Chemistry, Wayne State University, Detroit, Michigan 48202.
Pruess-Schwartz
D
D
Marnett
L J
LJ
D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide / Q000737:chemistry / Q000302:isolation & purification*; D000255:Adenosine Triphosphate / Q000138:chemical synthesis; D000818:Animals; D001345:Autoradiography; D001564:Benzo(a)pyrene / Q000737:chemistry; D002855:Chromatography, Thin Layer; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification*; D018736:DNA Adducts; D004101:Dihydroxydihydrobenzopyrenes / Q000737:chemistry; D005260:Female; D007553:Isotope Labeling; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009711:Nucleotides / Q000737:chemistry; D010761:Phosphorus Radioisotopes; D051381:Rats; D011919:Rats, Inbred Strains; D013237:Stereoisomerism
D018736:DNA Adducts; D004101:Dihydroxydihydrobenzopyrenes; D009711:Nucleotides; D010761:Phosphorus Radioisotopes; C043931:benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; C017228:benzo(a)pyrene 7,8-dihydrodiol; D001564:Benzo(a)pyrene; D015123:7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; D000255:Adenosine Triphosphate; D004247:DNA