Identification of (+) and (−) anti benzo[a]pyrene dihydrodiol epoxide–nucleic acid adducts by the 32P-postlabeling assay

Canella, Karen A.; Peltonen, Kimmo; Dipple, Anthony

doi:10.1093/carcin/12.6.1109

Cited by 36 publications

(27 citation statements)

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“…noted the presence of uncharacterized products in their investigation of the synthesis of base adducts by the reaction of B[a]PDE with 2′-deoxynucleotides. Two early eluting products were detected following HPLC-UV analysis of a B[a]PDE plus 2′-deoxyadenosine 5′-monophosphate reaction mixture that were resistant to hydrolysis by alkaline phosphatase unlike the base adducts (36). The identity of these products was not elucidated but it was concluded that their structure contained a nucleotide component that required the presence of 2′-deoxynucleotides in the B[a]PDE reaction mixture for their formation (36).…”

Section: Discussionmentioning

confidence: 99%

Detection of phosphodiester adducts formed by the reaction of benzo[a]pyrene diol epoxide with 2'-deoxynucleotides using collision-induced dissociation electrospray ionization tandem mass spectrometry

Gaskell

Kaur

Farmer

et al. 2007

Nucleic Acids Research

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In this study, we investigated the products formed following the reaction of benzo[a]pyrene-7,8-dihydrodiol-9,10-epoxide (B[a]PDE) with 2′-deoxynucleoside 3′-monophosphates. The B[a]PDE plus 2′-deoxynucleotide reaction mixtures were purified using solid phase extraction (SPE) and subjected to HPLC with fluorescence detection. Fractions corresponding to reaction product peaks were collected and desalted using SPE prior to analysis for the presence of molecular ions corresponding to m/z 648, 632, 608 and 623 [M−H]− consistent with B[a]PDE adducted (either on the base or phosphate group) 2′-deoxynucleotides of guanine, adenine, cytosine and thymine, respectively, using LC-ESI-MS/MS collision-induced dissociation (CID). Reaction products were identified having CID product ion spectra containing product ions at m/z 452, 436 and 412 [(B[a]Ptriol+base)−H]−, resulting from cleavage of the glycosidic bond between the 2′-deoxyribose and base, corresponding to B[a]PDE adducts of guanine, adenine and cytosine, respectively. Further reaction products were identified having unique CID product ion spectra characteristic of B[a]PDE adduct formation with the phosphate group of the 2′-deoxynucleotide. The presence of product ions at m/z 399 and 497 were observed for all four 2′-deoxynucleotides, corresponding to [(B[a]Ptriol+phosphate)−H]− and [(2′-deoxyribose+phosphate+B[a]Ptriol)−H]−, respectively. In conclusion, this investigation provides the first direct evidence for the formation of phosphodiester adducts by B[a]PDE following reaction with 2′-deoxynucleotides.

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Section: Discussionmentioning

confidence: 99%

Detection of phosphodiester adducts formed by the reaction of benzo[a]pyrene diol epoxide with 2'-deoxynucleotides using collision-induced dissociation electrospray ionization tandem mass spectrometry

Gaskell

Kaur

Farmer

et al. 2007

Nucleic Acids Research

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“…UV absorbance was monitored at 258 and 333 nm. Structure (cis or trans) and stereochemistry were confirmed by comparison with literature data (18,23,24). 15 N 6 -labeled 2′-deoxyadenosine (10 mg, 0.04 mmol) was dissolved completely in 1 mL of 50 mM Tris-HCl (pH 7.2) at 42°C.…”

Section: Scheme 2 Reaction Of Styrene Oxide With Adenosinementioning

confidence: 91%

“…The PAH dihydrodiol epoxides with which mechanisms were explored are shown in Scheme 5. Reactions of the epoxides with deoxyadenosine were carried out under conditions previously employed by Canella et al (24), except that 2′-deoxyadenosine was used instead of the 3′-phosphate (Scheme 6). The buffer was 50 mM Tris-HCl (pH 7.2).…”

Section: Reaction Of (R)-styrene Oxide With the [ 15 N 6 ]-Dado 24-mementioning

confidence: 99%

Studies of the Mechanisms of Adduction of 2‘-Deoxyadenosine with Styrene Oxide and Polycyclic Aromatic Hydrocarbon Dihydrodiol Epoxides

Kim

Finneman

Harris

et al. 2000

Chem. Res. Toxicol.

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The mechanism of adduction of 2'-deoxyadenosine by styrene oxide and polycyclic aromatic hydrocarbon dihydrodiol epoxides has been explored using (15)N(6)-labeled adenine nucleosides. The extent of reaction at N1 versus N(6) was evaluated by (1)H NMR of the N(6) adducts after allowing Dimroth rearrangement to occur. Products arising from attack at N1 followed by Dimroth rearrangement exhibited a small two-bond (1)H-(15)N coupling constant (N1-H2 J approximately 13 Hz); products from direct attack exhibited a much larger one-bond (1)H-(15)N coupling constant (J approximately 90 Hz). In the case of styrene oxide, all of the N(6) beta adduct arose by initial attack at N1, whereas the majority (70-80%) of the N(6) alpha adducts came from direct attack. The styrene oxide reaction was also studied with a self-complementary oligodeoxynucleotide (24-mer) containing nine (15)N(6)-labeled adenine residues. NMR examination of the N(6) alpha- and beta-styrene oxide adducts isolated after enzymatic degradation of the 24-mer gave very similar results, indicating that N1 attack can occur readily even with a duplexed oligonucleotide. With the PAH dihydrodiol epoxides, only naphthalene dihydrodiol epoxide exhibited significant initial reaction at N1 (50%). No detectable rearranged product was seen in reactions with benzo[a]pyrene dihydrodiol epoxide or non-bay or bay region benz[a]anthracene dihydrodiol epoxide; interestingly, a small amount of N1 attack (5-7%) was seen in the case of benzo[c]phenanthrene dihydrodiol epoxide. It appears that initial attack at N1 is only a significant reaction pathway for epoxides attached to a single aromatic ring.

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“…Modified 3'-phosphate nucleotides were prepared following a procedure described by Canella et al, [1991]. In brief, 75 p1 of each Bb]F dihydrodiol epoxide solution (9.5 pg/pl DMSO) was added to a I ml solution of either deoxyadenosine-3 '-phosphate or deoxyguanosine-3'-phosphate ( 10 mg nucleotide in 1 mlO.l M Tris buffer, pH 7.0).…”

Section: Preparation Of In Vitro Modified Nucleotidesmentioning

confidence: 99%

Mutagenic activity of the 4,5‐ and 9,10‐dihydrodiols of benzo[J]fluoranthene and their syn‐ and anti‐dihydrodiol epoxides in salmonella typhimurium

Marshall

Zhenmin

Weyand

et al. 1993

Environ and Mol Mutagen

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The objective of this study was to determine the relative mutagenic activities of the major dihydrodiol metabolites of benzo[j]fluoranthene (B[j]F) and their corresponding syn- and anti-dihydrodiol epoxides. Salmonella typhimurium tester strains TA97a, TA98, and TA100 were used to evaluate the mutagenic potencies of the parent hydrocarbon and these suspect proximate and ultimate mutagenic metabolites. B[j]F and the trans-dihydrodiol metabolites were active only in the presence of an external metabolic activation system (S9) with the exception of the B[j]F-4,5-diol, which was weakly active in TA98 and TA100 in the absence of S9. The B[j]F-4,5-diol was more mutagenic than the B[j]F-9,10-diol in tester strains TA98 and TA100, whereas the opposite effect was observed in TA97a. In the absence of S9, the anti-B[j]F-4,5-diol epoxide was more mutagenic than the syn-B[j]F-4,5-diol epoxide and the syn- and anti-B[j]F-9,10-diol epoxides in tester strains TA97a and TA100. The exceptional mutagenic potency of the anti-B[j]F-4,5-diol epoxide in TA100 resembles that observed by epoxides located within a fjord, or by the anti-diol epoxides of bay region methylated polycyclic aromatic hydrocarbons. In contrast, the mutagenicity of the pseudo bay region dihydrodiol epoxides arising from the B[j]F-9,10-diol more closely resembles that observed with the classical bay region dihydrodiol epoxides of chrysene. In summary, both dihydrodiol metabolites of B[j]F are mutagenic in S. typhimurium, and the relative potency varies among the tester strains. The highest mutagenic response was achieved in tester strain TA100, which detects base-pair substitutions. The most potent direct-acting dihydrodiol epoxide in this tester strain was the anti-B[j]F-4,5-diol epoxide, which agrees with the results of mouse skin painting studies that indicate that the B[j]F-4,5-diol is more tumorigenic that the parent hydrocarbon or the B[j]F-9,10-diol. A covalent DNA adduct formed between the anti-B[j]F-4,5-diol epoxide and deoxyguanosine was the major species of DNA adduct formed in S. typhimurium. This adduct corresponds to the major DNA adduct formed in mouse skin following application B[j]F.

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Identification of (+) and (−) anti benzo[a]pyrene dihydrodiol epoxide–nucleic acid adducts by the ³²P-postlabeling assay

Cited by 36 publications

References 0 publications

Detection of phosphodiester adducts formed by the reaction of benzo[a]pyrene diol epoxide with 2'-deoxynucleotides using collision-induced dissociation electrospray ionization tandem mass spectrometry

Detection of phosphodiester adducts formed by the reaction of benzo[a]pyrene diol epoxide with 2'-deoxynucleotides using collision-induced dissociation electrospray ionization tandem mass spectrometry

Studies of the Mechanisms of Adduction of 2‘-Deoxyadenosine with Styrene Oxide and Polycyclic Aromatic Hydrocarbon Dihydrodiol Epoxides

Mutagenic activity of the 4,5‐ and 9,10‐dihydrodiols of benzo[J]fluoranthene and their syn‐ and anti‐dihydrodiol epoxides in salmonella typhimurium

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