A Proteomic Study on the Toxicity of Arylamine Mixtures in Fisher-344 Rats

Arylamines are polyaromatic hydrocarbons (PAHs) which are by products in fuel burning, cooking and tobacco smoke. Health risks linked to arylamines include carcinogenicity, mutagenicity and teratogenicity. The current study examines the effect of arylamine mixtures (2-aminoanthracene (2AA), 2-aminofluorene (2AF), 1-aminopyrene (1AP)) exposure on Fisher-344 (F-344) rats using proteomics approach. Recently there has been an increasing interest in employing proteomics to study protein structure and function. Proteomics provides the opportunity to directly quantify the effect of toxicants on proteins. Thirty post-weaning 3-4 week old F-344 male rats were fed diets containing 0 µM and 50 µM of 2AA, 2AF, 1AP, 2AA/2AF, 2AA/1AP, 2AF/1AP and 2AA/2AF/1AP for 28 days. This was followed by total protein extraction from liver and pancreas tissues. Extracted protein samples are separated on a one dimensional gel and the bands cut, digested with trypsin for further separation and analysis on nano LC/MS/MS. The data generated from nano LC/MS/MS is searched using Mascot and quantified via spectral counting. Taking advantage of the fact that proteomics provides opportunity to study post-translational modifications, using a variety of artifactual examination; oxidation and deamidation modifications will be compared with other protein modifications in all treatment groups. Functional annotation and biological term enrichment analysis will be performed using DAVID Bioinformatics Resources. Functional pathway and gene network analysis of dysregulated proteins will be analyzed using Ariadne Pathway Studio. We anticipate finalizing the research soon and believe it will help in the development of biomarkers for the exposure of arlyamines mixtures.