Cytochrome c adducts with PCB quinoid metabolites

Miao Li, Lynn M. Teesch, Daryl J. Murry, R. Marshal Pope, Yalan Li, Larry W. Robertson, Gabriele Ludewig

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Polychlorinated biphenyls (PCBs) are a group of 209 individual congeners widely used as industrial chemicals. PCBs are found as by-products in dye and paint manufacture and are legacy, ubiquitous, and persistent as human and environmental contaminants. PCBs with fewer chlorine atoms may be metabolized to hydroxy- and dihydroxy-metabolites and further oxidized to quinoid metabolites both in vitro and in vivo. Specifically, quinoid metabolites may form adducts on nucleophilic sites within cells. We hypothesized that the PCB-quinones covalently bind to cytochrome c and, thereby, cause defects in the function of cytochrome c. In this study, synthetic PCB quinones, 2-(4′-chlorophenyl)-1,4-benzoquinone (PCB3-pQ), 4-4'-chlorophenyl)-1,2-benzoquinone (PCB3-oQ), 2-(3′, 5′-dichlorophenyl)-1,4-benzoquinone, 2-(3′,4′, 5′-trichlorophenyl)-1,4-benzoquinone, and 2-(4′-chlorophenyl)-3,6-dichloro-1,4-benzoquinone, were incubated with cytochrome c, and adducts were detected by liquid chromatography-mass spectrometry (LC-MS) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was employed to separate the adducted proteins, while trypsin digestion and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were applied to identify the amino acid binding sites on cytochrome c. Conformation change of cytochrome c after binding with PCB3-pQ was investigated by SYBYL-X simulation and cytochrome c function was examined. We found that more than one molecule of PCB-quinone may bind to one molecule of cytochrome c. Lysine and glutamic acid were identified as the predominant binding sites. Software simulation showed conformation changes of adducted cytochrome c. Additionally, cross-linking of cytochrome c was observed on the SDS-PAGE gel. Cytochrome c was found to lose its function as electron acceptor after incubation with PCB quinones. These data provide evidence that the covalent binding of PCB quinone metabolites to cytochrome c may be included among the toxic effects of PCBs.

Original languageEnglish (US)
Pages (from-to)2148-2159
Number of pages12
JournalEnvironmental Science and Pollution Research
Volume23
Issue number3
DOIs
StatePublished - Feb 1 2016

Fingerprint

Polychlorinated Biphenyls
Metabolites
Cytochromes c
Polychlorinated biphenyls
cytochrome
PCB
metabolite
Proteins
Quinones
Mass spectrometry
Liquid chromatography
mass spectrometry
Liquid Chromatography
liquid chromatography
gel
Gels
Electrophoresis
Sodium dodecyl sulfate
Binding sites
Sodium Dodecyl Sulfate

Keywords

  • 4-Chlorobiphenyl
  • Cytochrome c
  • Post-translational modification
  • Protein adducts
  • Protein cross-linking
  • Quinoid metabolites

ASJC Scopus subject areas

  • Environmental Chemistry
  • Pollution
  • Health, Toxicology and Mutagenesis

Cite this

Li, M., Teesch, L. M., Murry, D. J., Pope, R. M., Li, Y., Robertson, L. W., & Ludewig, G. (2016). Cytochrome c adducts with PCB quinoid metabolites. Environmental Science and Pollution Research, 23(3), 2148-2159. https://doi.org/10.1007/s11356-015-4801-3

Cytochrome c adducts with PCB quinoid metabolites. / Li, Miao; Teesch, Lynn M.; Murry, Daryl J.; Pope, R. Marshal; Li, Yalan; Robertson, Larry W.; Ludewig, Gabriele.

In: Environmental Science and Pollution Research, Vol. 23, No. 3, 01.02.2016, p. 2148-2159.

Research output: Contribution to journalArticle

Li, M, Teesch, LM, Murry, DJ, Pope, RM, Li, Y, Robertson, LW & Ludewig, G 2016, 'Cytochrome c adducts with PCB quinoid metabolites', Environmental Science and Pollution Research, vol. 23, no. 3, pp. 2148-2159. https://doi.org/10.1007/s11356-015-4801-3
Li, Miao ; Teesch, Lynn M. ; Murry, Daryl J. ; Pope, R. Marshal ; Li, Yalan ; Robertson, Larry W. ; Ludewig, Gabriele. / Cytochrome c adducts with PCB quinoid metabolites. In: Environmental Science and Pollution Research. 2016 ; Vol. 23, No. 3. pp. 2148-2159.
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AU - Murry, Daryl J.

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AU - Robertson, Larry W.

AU - Ludewig, Gabriele

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N2 - Polychlorinated biphenyls (PCBs) are a group of 209 individual congeners widely used as industrial chemicals. PCBs are found as by-products in dye and paint manufacture and are legacy, ubiquitous, and persistent as human and environmental contaminants. PCBs with fewer chlorine atoms may be metabolized to hydroxy- and dihydroxy-metabolites and further oxidized to quinoid metabolites both in vitro and in vivo. Specifically, quinoid metabolites may form adducts on nucleophilic sites within cells. We hypothesized that the PCB-quinones covalently bind to cytochrome c and, thereby, cause defects in the function of cytochrome c. In this study, synthetic PCB quinones, 2-(4′-chlorophenyl)-1,4-benzoquinone (PCB3-pQ), 4-4'-chlorophenyl)-1,2-benzoquinone (PCB3-oQ), 2-(3′, 5′-dichlorophenyl)-1,4-benzoquinone, 2-(3′,4′, 5′-trichlorophenyl)-1,4-benzoquinone, and 2-(4′-chlorophenyl)-3,6-dichloro-1,4-benzoquinone, were incubated with cytochrome c, and adducts were detected by liquid chromatography-mass spectrometry (LC-MS) and matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI TOF). Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was employed to separate the adducted proteins, while trypsin digestion and liquid chromatography-tandem mass spectrometry (LC-MS/MS) were applied to identify the amino acid binding sites on cytochrome c. Conformation change of cytochrome c after binding with PCB3-pQ was investigated by SYBYL-X simulation and cytochrome c function was examined. We found that more than one molecule of PCB-quinone may bind to one molecule of cytochrome c. Lysine and glutamic acid were identified as the predominant binding sites. Software simulation showed conformation changes of adducted cytochrome c. Additionally, cross-linking of cytochrome c was observed on the SDS-PAGE gel. Cytochrome c was found to lose its function as electron acceptor after incubation with PCB quinones. These data provide evidence that the covalent binding of PCB quinone metabolites to cytochrome c may be included among the toxic effects of PCBs.

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KW - 4-Chlorobiphenyl

KW - Cytochrome c

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KW - Protein adducts

KW - Protein cross-linking

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