Evaluation of Cysteine Adduct Formation in Rat Hemoglobin by 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and Related Compounds

Steven G. Carmella, Shashi S. Kagan, Thomas Spratt, Stephen S. Hecht

Research output: Contribution to journalArticle

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Abstract

The tobacco-specific nitrosamine 4-(methylnitrosamino)-l-(3-pyridyl)-1-butanone (NNK) forms hemoglobin adducts in rats. Upon mild base treatment, 4-hydroxy-l-(3-pyridyl)-l-butanone (HPB) is released from this globin. HPB has been suggested as a dosimeter for exposure to and metabolic activation of tobacco-specific nitrosamines. The purpose of this study was 2-fold: (a) to determine whether cysteine adducts of NNK were precursors to HPB, and (A) to determine to what extent cysteine adducts accounted for the material bound to globin that is not released upon mild base hydrolysis. The chemistry of cysteine adduct formation was investigated by reacting N-acetyl-L-cysteine with three model compounds for pyridyloxobutylation by metabolically activated NNK: 4-(carbethoxyni-trosamino)-l-(3-pyridyl)-l-butanone 0); 4-oxo-4-(3-pyridyl)-l-butyl-methanesulfonate (2); and 4-iodo-l-(3-pyridyl)-l-butanone (3). Five adducts were isolated and characterized by their spectral properties and by independent syntheses: two diastereomers of N-acetyl-5-|l-methyl-3-oxo-3-(3-pyridyl)propyl)-L-cysteine (7a,b), N-acetyI-5-[4-oxo-4-(3-pyridyl)-l-butyl]-L-cysteine (9), and two diastereomers of N-acetyl-5-{2-[2-(3-pyri-dyl)]-2,3,4,5-tetrahydrofuranyl}-L-cysteine (lla,b). Only Ua,b produced HPB upon mild base treatment; however, the chemistry of this adduct did not support its role as a major precursor to HPB released upon base treatment of globin. The formation of adducts in rat hemoglobin was then examined by reacting it with tritium-labeled 1 ([5–3H]1) or tritium-labeled 4-oxo-4-(3-pyridyl)-l-butyl p-toluenesulfonate ([5–3HJ4). The results demonstrated that the amino acids corresponding to 7a,b were present in hemoglobin reacted with [5–3H]l, accounting for 72% of the bound tritium. Amino acids corresponding to 9 were not detected in this globin. In contrast, hemoglobin reacted with [5–3H]4 contained the amino acid corresponding to 9 (15% of bound tritium), but not those corresponding to 7a,b. These results indicated that the αβ-unsaturated ketone, l-(3-pyridyl)-2-buten-l-one (5), played a major role in the hemoglobin binding of 1, but not of 4. Cysteine adducts were not detected in globin isolated from rats treated with [5–3H]NNK. The results of this study provide insights into the mechanisms of cysteine adduct formation in vitro by pyridyloxobutylating agents and indicate that;these adducts are not formed in NNK-treated rats.

Original languageEnglish (US)
Pages (from-to)5453-5459
Number of pages7
JournalCancer Research
Volume50
Issue number17
StatePublished - Sep 1 1990

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Cysteine
Hemoglobins
Globins
Butanones
Tritium
Nitrosamines
Amino Acids
Tobacco
4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone
Acetylcysteine
Ketones
Hydrolysis

All Science Journal Classification (ASJC) codes

  • Cancer Research
  • Oncology

Cite this

@article{42fee3fd27a04048ae39277132ed77dd,
title = "Evaluation of Cysteine Adduct Formation in Rat Hemoglobin by 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and Related Compounds",
abstract = "The tobacco-specific nitrosamine 4-(methylnitrosamino)-l-(3-pyridyl)-1-butanone (NNK) forms hemoglobin adducts in rats. Upon mild base treatment, 4-hydroxy-l-(3-pyridyl)-l-butanone (HPB) is released from this globin. HPB has been suggested as a dosimeter for exposure to and metabolic activation of tobacco-specific nitrosamines. The purpose of this study was 2-fold: (a) to determine whether cysteine adducts of NNK were precursors to HPB, and (A) to determine to what extent cysteine adducts accounted for the material bound to globin that is not released upon mild base hydrolysis. The chemistry of cysteine adduct formation was investigated by reacting N-acetyl-L-cysteine with three model compounds for pyridyloxobutylation by metabolically activated NNK: 4-(carbethoxyni-trosamino)-l-(3-pyridyl)-l-butanone 0); 4-oxo-4-(3-pyridyl)-l-butyl-methanesulfonate (2); and 4-iodo-l-(3-pyridyl)-l-butanone (3). Five adducts were isolated and characterized by their spectral properties and by independent syntheses: two diastereomers of N-acetyl-5-|l-methyl-3-oxo-3-(3-pyridyl)propyl)-L-cysteine (7a,b), N-acetyI-5-[4-oxo-4-(3-pyridyl)-l-butyl]-L-cysteine (9), and two diastereomers of N-acetyl-5-{2-[2-(3-pyri-dyl)]-2,3,4,5-tetrahydrofuranyl}-L-cysteine (lla,b). Only Ua,b produced HPB upon mild base treatment; however, the chemistry of this adduct did not support its role as a major precursor to HPB released upon base treatment of globin. The formation of adducts in rat hemoglobin was then examined by reacting it with tritium-labeled 1 ([5–3H]1) or tritium-labeled 4-oxo-4-(3-pyridyl)-l-butyl p-toluenesulfonate ([5–3HJ4). The results demonstrated that the amino acids corresponding to 7a,b were present in hemoglobin reacted with [5–3H]l, accounting for 72{\%} of the bound tritium. Amino acids corresponding to 9 were not detected in this globin. In contrast, hemoglobin reacted with [5–3H]4 contained the amino acid corresponding to 9 (15{\%} of bound tritium), but not those corresponding to 7a,b. These results indicated that the αβ-unsaturated ketone, l-(3-pyridyl)-2-buten-l-one (5), played a major role in the hemoglobin binding of 1, but not of 4. Cysteine adducts were not detected in globin isolated from rats treated with [5–3H]NNK. The results of this study provide insights into the mechanisms of cysteine adduct formation in vitro by pyridyloxobutylating agents and indicate that;these adducts are not formed in NNK-treated rats.",
author = "Carmella, {Steven G.} and Kagan, {Shashi S.} and Thomas Spratt and Hecht, {Stephen S.}",
year = "1990",
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Evaluation of Cysteine Adduct Formation in Rat Hemoglobin by 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and Related Compounds. / Carmella, Steven G.; Kagan, Shashi S.; Spratt, Thomas; Hecht, Stephen S.

In: Cancer Research, Vol. 50, No. 17, 01.09.1990, p. 5453-5459.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Evaluation of Cysteine Adduct Formation in Rat Hemoglobin by 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone and Related Compounds

AU - Carmella, Steven G.

AU - Kagan, Shashi S.

AU - Spratt, Thomas

AU - Hecht, Stephen S.

PY - 1990/9/1

Y1 - 1990/9/1

N2 - The tobacco-specific nitrosamine 4-(methylnitrosamino)-l-(3-pyridyl)-1-butanone (NNK) forms hemoglobin adducts in rats. Upon mild base treatment, 4-hydroxy-l-(3-pyridyl)-l-butanone (HPB) is released from this globin. HPB has been suggested as a dosimeter for exposure to and metabolic activation of tobacco-specific nitrosamines. The purpose of this study was 2-fold: (a) to determine whether cysteine adducts of NNK were precursors to HPB, and (A) to determine to what extent cysteine adducts accounted for the material bound to globin that is not released upon mild base hydrolysis. The chemistry of cysteine adduct formation was investigated by reacting N-acetyl-L-cysteine with three model compounds for pyridyloxobutylation by metabolically activated NNK: 4-(carbethoxyni-trosamino)-l-(3-pyridyl)-l-butanone 0); 4-oxo-4-(3-pyridyl)-l-butyl-methanesulfonate (2); and 4-iodo-l-(3-pyridyl)-l-butanone (3). Five adducts were isolated and characterized by their spectral properties and by independent syntheses: two diastereomers of N-acetyl-5-|l-methyl-3-oxo-3-(3-pyridyl)propyl)-L-cysteine (7a,b), N-acetyI-5-[4-oxo-4-(3-pyridyl)-l-butyl]-L-cysteine (9), and two diastereomers of N-acetyl-5-{2-[2-(3-pyri-dyl)]-2,3,4,5-tetrahydrofuranyl}-L-cysteine (lla,b). Only Ua,b produced HPB upon mild base treatment; however, the chemistry of this adduct did not support its role as a major precursor to HPB released upon base treatment of globin. The formation of adducts in rat hemoglobin was then examined by reacting it with tritium-labeled 1 ([5–3H]1) or tritium-labeled 4-oxo-4-(3-pyridyl)-l-butyl p-toluenesulfonate ([5–3HJ4). The results demonstrated that the amino acids corresponding to 7a,b were present in hemoglobin reacted with [5–3H]l, accounting for 72% of the bound tritium. Amino acids corresponding to 9 were not detected in this globin. In contrast, hemoglobin reacted with [5–3H]4 contained the amino acid corresponding to 9 (15% of bound tritium), but not those corresponding to 7a,b. These results indicated that the αβ-unsaturated ketone, l-(3-pyridyl)-2-buten-l-one (5), played a major role in the hemoglobin binding of 1, but not of 4. Cysteine adducts were not detected in globin isolated from rats treated with [5–3H]NNK. The results of this study provide insights into the mechanisms of cysteine adduct formation in vitro by pyridyloxobutylating agents and indicate that;these adducts are not formed in NNK-treated rats.

AB - The tobacco-specific nitrosamine 4-(methylnitrosamino)-l-(3-pyridyl)-1-butanone (NNK) forms hemoglobin adducts in rats. Upon mild base treatment, 4-hydroxy-l-(3-pyridyl)-l-butanone (HPB) is released from this globin. HPB has been suggested as a dosimeter for exposure to and metabolic activation of tobacco-specific nitrosamines. The purpose of this study was 2-fold: (a) to determine whether cysteine adducts of NNK were precursors to HPB, and (A) to determine to what extent cysteine adducts accounted for the material bound to globin that is not released upon mild base hydrolysis. The chemistry of cysteine adduct formation was investigated by reacting N-acetyl-L-cysteine with three model compounds for pyridyloxobutylation by metabolically activated NNK: 4-(carbethoxyni-trosamino)-l-(3-pyridyl)-l-butanone 0); 4-oxo-4-(3-pyridyl)-l-butyl-methanesulfonate (2); and 4-iodo-l-(3-pyridyl)-l-butanone (3). Five adducts were isolated and characterized by their spectral properties and by independent syntheses: two diastereomers of N-acetyl-5-|l-methyl-3-oxo-3-(3-pyridyl)propyl)-L-cysteine (7a,b), N-acetyI-5-[4-oxo-4-(3-pyridyl)-l-butyl]-L-cysteine (9), and two diastereomers of N-acetyl-5-{2-[2-(3-pyri-dyl)]-2,3,4,5-tetrahydrofuranyl}-L-cysteine (lla,b). Only Ua,b produced HPB upon mild base treatment; however, the chemistry of this adduct did not support its role as a major precursor to HPB released upon base treatment of globin. The formation of adducts in rat hemoglobin was then examined by reacting it with tritium-labeled 1 ([5–3H]1) or tritium-labeled 4-oxo-4-(3-pyridyl)-l-butyl p-toluenesulfonate ([5–3HJ4). The results demonstrated that the amino acids corresponding to 7a,b were present in hemoglobin reacted with [5–3H]l, accounting for 72% of the bound tritium. Amino acids corresponding to 9 were not detected in this globin. In contrast, hemoglobin reacted with [5–3H]4 contained the amino acid corresponding to 9 (15% of bound tritium), but not those corresponding to 7a,b. These results indicated that the αβ-unsaturated ketone, l-(3-pyridyl)-2-buten-l-one (5), played a major role in the hemoglobin binding of 1, but not of 4. Cysteine adducts were not detected in globin isolated from rats treated with [5–3H]NNK. The results of this study provide insights into the mechanisms of cysteine adduct formation in vitro by pyridyloxobutylating agents and indicate that;these adducts are not formed in NNK-treated rats.

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