Abstract
Benzo[a]pyrene (B[a]P) is an environmental contaminant mainly studied for its toxic/carcinogenic effects. For a comprehensive and pathway orientated mechanistic understanding of the effects directly triggered by a toxic (5 μM) or a subtoxic (50 nM) concentration of B[a]P or indirectly by its metabolites, we conducted time series experiments for up to 24 h to study the effects in murine hepatocytes. These cells rapidly take up and actively metabolize B[a]P, which was followed by quantitative analysis of the concentration of intracellular B[a]P and seven representative degradation products. Exposure with 5 μM B[a]P led to a maximal intracellular concentration of 1604 pmol/5 × 104 cells, leveling at 55 pmol/5 × 104 cells by the end of the time course. Changes in the global proteome (>1000 protein profiles) and metabolome (163 metabolites) were assessed in combination with B[a]P degradation. Abundance profiles of 236 (both concentrations), 190 (only 5 μM), and 150 (only 50 nM) proteins were found to be regulated in response to B[a]P in a time-dependent manner. At the endogenous metabolite level amino acids, acylcarnitines and glycerophospholipids were particularly affected by B[a]P. The comprehensive chemical, proteome and metabolomic data enabled the identification of effects on the pathway level in a time-resolved manner. So in addition to known alterations, also protein synthesis, lipid metabolism, and membrane dysfunction were identified as B[a]P specific effects.
Original language | English |
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Journal | Journal of Proteome Research |
Volume | 14 |
Issue number | 1 |
Pages (from-to) | 164-182 |
Number of pages | 19 |
ISSN | 1535-3893 |
DOIs | |
Publication status | Published - 1 Jan 2015 |
Keywords
- aryl hydrocarbon receptor (Ahr)
- Benzo[ a ]pyrene (B[ a ]P)
- B[ a ]P metabolites
- Hepa1c1c7 cells
- metabolomics
- oxidative stress
- protein expression analysis
- proteomics
- SILAC