Non-enzymatic ATP-mediated binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA

Eleanor G Rogan, R. W. Roth, Patricia A. Katomski-Beck, Jeanne R. Laubscher, Ercole Cavalieri

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

ATP mediates covalent binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA. This non-enzymatic reaction has been studied with 6-[14C]hydroxymethylbenzo[a]pyrene ([14C]BP-6-CH2OH) and 7-[14C]hydroxymethylbenz[a]anthracene ([14C]BA-7-CH2OH) at 37°C in Tris buffer (pH 7.0). While ADP mediates the reaction 25-50% as well as ATP, six other possible phosphate donors including AMP were inactive as cofactors. A complex response to ATP occurred in which low binding of BP-6-CH2OH or BA-7-CH2OH was observed at concentrations of ATP below 2.5 mM, but a greater than linear response to higher concentrations of ATP was observed until ATP was saturating. Binding of the substrates to RNA was much lower than to DNA. Fluorescence spectra of BP-6-CH2OH bound to DNA were almost identical to the spectra of 6-bromomethylbenzo[a]pyrene bound to DNA and free 6-methylbenzo[a]pyrene, indicating that ATP-mediated binding of BP-6-CH2OH to DNA occurs at the 6-methyl group. The fate of ATP and ADP in the binding reaction of BP-6-CH2OH was examined by thin layer chromatography. Loss of one phosphate group occurs during the reaction. With ATP the rate of loss is about 100-fold greater than the rate of binding of BP-6-CH2OH to DNA. This implies that the binding reaction proceeds through formation of a presumed reactive and unstable phosphate ester intermediate which then inefficiently binds to DNA.

Original languageEnglish (US)
Pages (from-to)51-63
Number of pages13
JournalChemico-Biological Interactions
Volume31
Issue number1
DOIs
StatePublished - Jul 1980

Fingerprint

Aromatic Hydrocarbons
Adenosine Triphosphate
Derivatives
DNA
Phosphates
Adenosine Diphosphate
Thin layer chromatography
Tromethamine
Adenosine Monophosphate
Thin Layer Chromatography
Esters
Fluorescence
RNA

ASJC Scopus subject areas

  • Toxicology

Cite this

Non-enzymatic ATP-mediated binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA. / Rogan, Eleanor G; Roth, R. W.; Katomski-Beck, Patricia A.; Laubscher, Jeanne R.; Cavalieri, Ercole.

In: Chemico-Biological Interactions, Vol. 31, No. 1, 07.1980, p. 51-63.

Research output: Contribution to journalArticle

Rogan, Eleanor G ; Roth, R. W. ; Katomski-Beck, Patricia A. ; Laubscher, Jeanne R. ; Cavalieri, Ercole. / Non-enzymatic ATP-mediated binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA. In: Chemico-Biological Interactions. 1980 ; Vol. 31, No. 1. pp. 51-63.
@article{b395162b47724b96a032ad27ff3ff9da,
title = "Non-enzymatic ATP-mediated binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA",
abstract = "ATP mediates covalent binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA. This non-enzymatic reaction has been studied with 6-[14C]hydroxymethylbenzo[a]pyrene ([14C]BP-6-CH2OH) and 7-[14C]hydroxymethylbenz[a]anthracene ([14C]BA-7-CH2OH) at 37°C in Tris buffer (pH 7.0). While ADP mediates the reaction 25-50{\%} as well as ATP, six other possible phosphate donors including AMP were inactive as cofactors. A complex response to ATP occurred in which low binding of BP-6-CH2OH or BA-7-CH2OH was observed at concentrations of ATP below 2.5 mM, but a greater than linear response to higher concentrations of ATP was observed until ATP was saturating. Binding of the substrates to RNA was much lower than to DNA. Fluorescence spectra of BP-6-CH2OH bound to DNA were almost identical to the spectra of 6-bromomethylbenzo[a]pyrene bound to DNA and free 6-methylbenzo[a]pyrene, indicating that ATP-mediated binding of BP-6-CH2OH to DNA occurs at the 6-methyl group. The fate of ATP and ADP in the binding reaction of BP-6-CH2OH was examined by thin layer chromatography. Loss of one phosphate group occurs during the reaction. With ATP the rate of loss is about 100-fold greater than the rate of binding of BP-6-CH2OH to DNA. This implies that the binding reaction proceeds through formation of a presumed reactive and unstable phosphate ester intermediate which then inefficiently binds to DNA.",
author = "Rogan, {Eleanor G} and Roth, {R. W.} and Katomski-Beck, {Patricia A.} and Laubscher, {Jeanne R.} and Ercole Cavalieri",
year = "1980",
month = "7",
doi = "10.1016/0009-2797(80)90138-6",
language = "English (US)",
volume = "31",
pages = "51--63",
journal = "Chemico-Biological Interactions",
issn = "0009-2797",
publisher = "Elsevier Ireland Ltd",
number = "1",

}

TY - JOUR

T1 - Non-enzymatic ATP-mediated binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA

AU - Rogan, Eleanor G

AU - Roth, R. W.

AU - Katomski-Beck, Patricia A.

AU - Laubscher, Jeanne R.

AU - Cavalieri, Ercole

PY - 1980/7

Y1 - 1980/7

N2 - ATP mediates covalent binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA. This non-enzymatic reaction has been studied with 6-[14C]hydroxymethylbenzo[a]pyrene ([14C]BP-6-CH2OH) and 7-[14C]hydroxymethylbenz[a]anthracene ([14C]BA-7-CH2OH) at 37°C in Tris buffer (pH 7.0). While ADP mediates the reaction 25-50% as well as ATP, six other possible phosphate donors including AMP were inactive as cofactors. A complex response to ATP occurred in which low binding of BP-6-CH2OH or BA-7-CH2OH was observed at concentrations of ATP below 2.5 mM, but a greater than linear response to higher concentrations of ATP was observed until ATP was saturating. Binding of the substrates to RNA was much lower than to DNA. Fluorescence spectra of BP-6-CH2OH bound to DNA were almost identical to the spectra of 6-bromomethylbenzo[a]pyrene bound to DNA and free 6-methylbenzo[a]pyrene, indicating that ATP-mediated binding of BP-6-CH2OH to DNA occurs at the 6-methyl group. The fate of ATP and ADP in the binding reaction of BP-6-CH2OH was examined by thin layer chromatography. Loss of one phosphate group occurs during the reaction. With ATP the rate of loss is about 100-fold greater than the rate of binding of BP-6-CH2OH to DNA. This implies that the binding reaction proceeds through formation of a presumed reactive and unstable phosphate ester intermediate which then inefficiently binds to DNA.

AB - ATP mediates covalent binding of hydroxymethyl derivatives of aromatic hydrocarbons to DNA. This non-enzymatic reaction has been studied with 6-[14C]hydroxymethylbenzo[a]pyrene ([14C]BP-6-CH2OH) and 7-[14C]hydroxymethylbenz[a]anthracene ([14C]BA-7-CH2OH) at 37°C in Tris buffer (pH 7.0). While ADP mediates the reaction 25-50% as well as ATP, six other possible phosphate donors including AMP were inactive as cofactors. A complex response to ATP occurred in which low binding of BP-6-CH2OH or BA-7-CH2OH was observed at concentrations of ATP below 2.5 mM, but a greater than linear response to higher concentrations of ATP was observed until ATP was saturating. Binding of the substrates to RNA was much lower than to DNA. Fluorescence spectra of BP-6-CH2OH bound to DNA were almost identical to the spectra of 6-bromomethylbenzo[a]pyrene bound to DNA and free 6-methylbenzo[a]pyrene, indicating that ATP-mediated binding of BP-6-CH2OH to DNA occurs at the 6-methyl group. The fate of ATP and ADP in the binding reaction of BP-6-CH2OH was examined by thin layer chromatography. Loss of one phosphate group occurs during the reaction. With ATP the rate of loss is about 100-fold greater than the rate of binding of BP-6-CH2OH to DNA. This implies that the binding reaction proceeds through formation of a presumed reactive and unstable phosphate ester intermediate which then inefficiently binds to DNA.

UR - http://www.scopus.com/inward/record.url?scp=0018890284&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0018890284&partnerID=8YFLogxK

U2 - 10.1016/0009-2797(80)90138-6

DO - 10.1016/0009-2797(80)90138-6

M3 - Article

C2 - 6156020

AN - SCOPUS:0018890284

VL - 31

SP - 51

EP - 63

JO - Chemico-Biological Interactions

JF - Chemico-Biological Interactions

SN - 0009-2797

IS - 1

ER -