Analysis for N2-(Pyridyloxobutyl)deoxyguanosine Adducts in DNA of Tissues Exposed to Tritium-Labeled 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and N'-Nitrosonornicotine

Thomas Spratt, Neil Trushin, Dorothy Lin, Stephen S. Hecht

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The tobacco-specific carcinogens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) are metabolically activated to DNA binding intermediates, partially via 4-(3-pyridyl)-4-oxobutanediazohydroxide (7) or related carbonium ions. Previous studies have shown that generation of 7 from 4-(carbethoxynitrosamino)-1-(3-pyridyl)-1-butanone (11) in the presence of deoxyguanosine yields a major adduct identified as 2'-deoxy-N-[1-methyl-3-oxo-3-(3-pyridyl)propyl]guanosine (adduct 1). These results suggested that adduct 1 should be present in DNA of tissues that can metabolically activate NNK and NNN. In the present study, we evaluate the formation of adduct 1 and its structurally related straight-chain analogue 2'-deoxy-N-[4-oxo-4-(3-pyridyl)butyl]guanosine (adduct 2) in DNA of tissues of rats treated with [5-3H]NNK or [5-3H]NNN, and in DNA of nasal mucosa that had been cultured in medium containing [5-3H]NNK or [5-3H]NNN. Hepatic DNA from rats treated with [5-3H]NNK was enzymatically hydrolyzed to deoxyribonucleosides and analyzed by HPLC. One of the radioactive peaks, peak E, coeluted with adduct 1. However, treatment of peak E with NaBH4 resulted in the formation of products different from those produced by NaBH4 treatment of adduct 1, demonstrating that adduct 1 could not be detected under these conditions. Hydrolysis of peak E with acid produced 4-hydroxy-1-(3-pyridyl)-1-butanone (9), suggesting that peak E might be adduct 2. Therefore, adduct 2 was synthesized by reaction of deoxyguanosine with 1-(3-pyridyl)butane-1,4-dione (5) in the presence of NaCNBH3. Its HPLC retention time, however, was different from that of peak E. Peak E was also detected in DNA of nasal mucosa incubated with [5-3H]NNK or [5-3H]NNN. The HPLC chromatograms obtained upon enzymatic hydrolysis of DNA exposed to [5-3H]NNK or [5-3H]NNN were generally similar, indicating the operation of a common alkylation pathway via diazohydroxide 7 or a related carbonium ion. This pathway does not however lead to detectable levels of adducts 1 or 2 in DNA (detection limit using [5-3H]NNK or [5-3H]NNN, approximately 0.05 pmol/mg of DNA).

Original languageEnglish (US)
Pages (from-to)169-173
Number of pages5
JournalChemical Research in Toxicology
Volume2
Issue number3
DOIs
StatePublished - May 1 1989

Fingerprint

N'-nitrosonornicotine
Deoxyguanosine
Tritium
DNA Adducts
Tissue
DNA
Nasal Mucosa
Guanosine
High Pressure Liquid Chromatography
pyrazine-2-diazohydroxide
Rats
Hydrolysis
Ions
Deoxyribonucleosides
4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone
Enzymatic hydrolysis
Tobacco
Alkylation

All Science Journal Classification (ASJC) codes

  • Toxicology

Cite this

@article{732ac91711304d04ac84f7f349d87626,
title = "Analysis for N2-(Pyridyloxobutyl)deoxyguanosine Adducts in DNA of Tissues Exposed to Tritium-Labeled 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and N'-Nitrosonornicotine",
abstract = "The tobacco-specific carcinogens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) are metabolically activated to DNA binding intermediates, partially via 4-(3-pyridyl)-4-oxobutanediazohydroxide (7) or related carbonium ions. Previous studies have shown that generation of 7 from 4-(carbethoxynitrosamino)-1-(3-pyridyl)-1-butanone (11) in the presence of deoxyguanosine yields a major adduct identified as 2'-deoxy-N-[1-methyl-3-oxo-3-(3-pyridyl)propyl]guanosine (adduct 1). These results suggested that adduct 1 should be present in DNA of tissues that can metabolically activate NNK and NNN. In the present study, we evaluate the formation of adduct 1 and its structurally related straight-chain analogue 2'-deoxy-N-[4-oxo-4-(3-pyridyl)butyl]guanosine (adduct 2) in DNA of tissues of rats treated with [5-3H]NNK or [5-3H]NNN, and in DNA of nasal mucosa that had been cultured in medium containing [5-3H]NNK or [5-3H]NNN. Hepatic DNA from rats treated with [5-3H]NNK was enzymatically hydrolyzed to deoxyribonucleosides and analyzed by HPLC. One of the radioactive peaks, peak E, coeluted with adduct 1. However, treatment of peak E with NaBH4 resulted in the formation of products different from those produced by NaBH4 treatment of adduct 1, demonstrating that adduct 1 could not be detected under these conditions. Hydrolysis of peak E with acid produced 4-hydroxy-1-(3-pyridyl)-1-butanone (9), suggesting that peak E might be adduct 2. Therefore, adduct 2 was synthesized by reaction of deoxyguanosine with 1-(3-pyridyl)butane-1,4-dione (5) in the presence of NaCNBH3. Its HPLC retention time, however, was different from that of peak E. Peak E was also detected in DNA of nasal mucosa incubated with [5-3H]NNK or [5-3H]NNN. The HPLC chromatograms obtained upon enzymatic hydrolysis of DNA exposed to [5-3H]NNK or [5-3H]NNN were generally similar, indicating the operation of a common alkylation pathway via diazohydroxide 7 or a related carbonium ion. This pathway does not however lead to detectable levels of adducts 1 or 2 in DNA (detection limit using [5-3H]NNK or [5-3H]NNN, approximately 0.05 pmol/mg of DNA).",
author = "Thomas Spratt and Neil Trushin and Dorothy Lin and Hecht, {Stephen S.}",
year = "1989",
month = "5",
day = "1",
doi = "10.1021/tx00009a008",
language = "English (US)",
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pages = "169--173",
journal = "Chemical Research in Toxicology",
issn = "0893-228X",
publisher = "American Chemical Society",
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}

Analysis for N2-(Pyridyloxobutyl)deoxyguanosine Adducts in DNA of Tissues Exposed to Tritium-Labeled 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and N'-Nitrosonornicotine. / Spratt, Thomas; Trushin, Neil; Lin, Dorothy; Hecht, Stephen S.

In: Chemical Research in Toxicology, Vol. 2, No. 3, 01.05.1989, p. 169-173.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Analysis for N2-(Pyridyloxobutyl)deoxyguanosine Adducts in DNA of Tissues Exposed to Tritium-Labeled 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and N'-Nitrosonornicotine

AU - Spratt, Thomas

AU - Trushin, Neil

AU - Lin, Dorothy

AU - Hecht, Stephen S.

PY - 1989/5/1

Y1 - 1989/5/1

N2 - The tobacco-specific carcinogens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) are metabolically activated to DNA binding intermediates, partially via 4-(3-pyridyl)-4-oxobutanediazohydroxide (7) or related carbonium ions. Previous studies have shown that generation of 7 from 4-(carbethoxynitrosamino)-1-(3-pyridyl)-1-butanone (11) in the presence of deoxyguanosine yields a major adduct identified as 2'-deoxy-N-[1-methyl-3-oxo-3-(3-pyridyl)propyl]guanosine (adduct 1). These results suggested that adduct 1 should be present in DNA of tissues that can metabolically activate NNK and NNN. In the present study, we evaluate the formation of adduct 1 and its structurally related straight-chain analogue 2'-deoxy-N-[4-oxo-4-(3-pyridyl)butyl]guanosine (adduct 2) in DNA of tissues of rats treated with [5-3H]NNK or [5-3H]NNN, and in DNA of nasal mucosa that had been cultured in medium containing [5-3H]NNK or [5-3H]NNN. Hepatic DNA from rats treated with [5-3H]NNK was enzymatically hydrolyzed to deoxyribonucleosides and analyzed by HPLC. One of the radioactive peaks, peak E, coeluted with adduct 1. However, treatment of peak E with NaBH4 resulted in the formation of products different from those produced by NaBH4 treatment of adduct 1, demonstrating that adduct 1 could not be detected under these conditions. Hydrolysis of peak E with acid produced 4-hydroxy-1-(3-pyridyl)-1-butanone (9), suggesting that peak E might be adduct 2. Therefore, adduct 2 was synthesized by reaction of deoxyguanosine with 1-(3-pyridyl)butane-1,4-dione (5) in the presence of NaCNBH3. Its HPLC retention time, however, was different from that of peak E. Peak E was also detected in DNA of nasal mucosa incubated with [5-3H]NNK or [5-3H]NNN. The HPLC chromatograms obtained upon enzymatic hydrolysis of DNA exposed to [5-3H]NNK or [5-3H]NNN were generally similar, indicating the operation of a common alkylation pathway via diazohydroxide 7 or a related carbonium ion. This pathway does not however lead to detectable levels of adducts 1 or 2 in DNA (detection limit using [5-3H]NNK or [5-3H]NNN, approximately 0.05 pmol/mg of DNA).

AB - The tobacco-specific carcinogens 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) and N'-nitrosonornicotine (NNN) are metabolically activated to DNA binding intermediates, partially via 4-(3-pyridyl)-4-oxobutanediazohydroxide (7) or related carbonium ions. Previous studies have shown that generation of 7 from 4-(carbethoxynitrosamino)-1-(3-pyridyl)-1-butanone (11) in the presence of deoxyguanosine yields a major adduct identified as 2'-deoxy-N-[1-methyl-3-oxo-3-(3-pyridyl)propyl]guanosine (adduct 1). These results suggested that adduct 1 should be present in DNA of tissues that can metabolically activate NNK and NNN. In the present study, we evaluate the formation of adduct 1 and its structurally related straight-chain analogue 2'-deoxy-N-[4-oxo-4-(3-pyridyl)butyl]guanosine (adduct 2) in DNA of tissues of rats treated with [5-3H]NNK or [5-3H]NNN, and in DNA of nasal mucosa that had been cultured in medium containing [5-3H]NNK or [5-3H]NNN. Hepatic DNA from rats treated with [5-3H]NNK was enzymatically hydrolyzed to deoxyribonucleosides and analyzed by HPLC. One of the radioactive peaks, peak E, coeluted with adduct 1. However, treatment of peak E with NaBH4 resulted in the formation of products different from those produced by NaBH4 treatment of adduct 1, demonstrating that adduct 1 could not be detected under these conditions. Hydrolysis of peak E with acid produced 4-hydroxy-1-(3-pyridyl)-1-butanone (9), suggesting that peak E might be adduct 2. Therefore, adduct 2 was synthesized by reaction of deoxyguanosine with 1-(3-pyridyl)butane-1,4-dione (5) in the presence of NaCNBH3. Its HPLC retention time, however, was different from that of peak E. Peak E was also detected in DNA of nasal mucosa incubated with [5-3H]NNK or [5-3H]NNN. The HPLC chromatograms obtained upon enzymatic hydrolysis of DNA exposed to [5-3H]NNK or [5-3H]NNN were generally similar, indicating the operation of a common alkylation pathway via diazohydroxide 7 or a related carbonium ion. This pathway does not however lead to detectable levels of adducts 1 or 2 in DNA (detection limit using [5-3H]NNK or [5-3H]NNN, approximately 0.05 pmol/mg of DNA).

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