Investigation of hepatotoxicity of acrylamide using mass spectrometry-based proteomics and N-glycoproteomics in mouse model.

Acrylamide is a widely used and distributed chemical proven to be a hepatotoxicant; however, the toxicological mechanisms on post-translational modifications regulation, which is most relevant to regulating the activity of proteins, are largely unknown. Here we report mass spectrometry-based toxico-proteomics and N-glycoproteomics study of AA using ICR mouse model with the exposure experimental group and the wild-type control group. 16,640 peptides from 2,956 proteins and 19,613 intact N-glycopeptides from 3,515 N-glycoproteins were identified in proteomics and N-glycoproteomics, where 99 differentially expressed proteins and 303 differentially expressed intact N-glycopeptides from 150 N-glycoproteins were quantified. Functional annotation of differentially expressed proteins and N-glycoproteins enables identification of AA hepatotoxicity pathways of autophagy, apoptosis and inflammation through stresses of reactive oxygen species and endoplasmic reticulum. 11 potential N-glycoprotein biomarkers were filtered through interactome and functional network analysis, and 5 (UGT1A9, CTSD, PSAP, CES1F and CD163) of them with more than 50 % frequency occurrence in parallel reaction monitoring verification experiment. These results provide a deep and comprehensive understanding of AA hepatotoxicity, especially from the perspective of N-glycoproteins; in addition, the integrated proteomics and N-glycoproteomics pipeline can be extended to the study of other toxicants.