Covalent Protein Modification: An Unignorable Factor for Bisphenol A‑Induced Hepatotoxicity

• Published in: Environmental science & technology Vol. 56; no. 13; pp. 9536 - 9545
• Main Authors: Hu, Xiaolan, Wu, Jian-Lin, Miao, Wen, Long, Fei, Pan, Hudan, Peng, Tao, Yao, Xiaojun, Li, Na
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 05.07.2022
• Subjects: Adducts; Amino acids; Biocompatibility; Bisphenol A; Catalase; Contaminants; Covalence; Cysteine; Ecotoxicology and Public Health; Glutathione; Hepatotoxicity; Incubation; Liquid chromatography; Mass spectrometry; Mass spectroscopy; Metabolites; Oxidative stress; Pollutants; Protein adducts; Proteins; Proteomics; Toxicity; Workflow; bisphenol A; reactive metabolites; LC−MS; hepatotoxicity; covalent protein modification
• ISSN: 0013-936X; 1520-5851
• DOI: 10.1021/acs.est.2c01307

Covalent modification of proteins by reactive pollutants/metabolites might trigger various toxicities resulting from the disruption of protein structures and/or functions, which is critical for understanding the mechanism of pollutants-induced toxicity. However, this mechanism has rarely been touched on due to the lack of a methodology. In this research, the protein modification of bisphenol A (BPA) in rats was characterized using a series of liquid chromatography–tandem mass spectrometry (LC–MS) approaches. BPA-modified cysteine (Cys1) was first released from proteins via enzymatic hydrolysis and identified using LC–MS. Moreover, the positive correlation between Cys1 and hepatotoxicity indicated the involvement of protein modification in BPA toxicity. Then, in vitro incubation of BPA with amino acids and protein confirmed that BPA could specifically modify cysteine residues of proteins after bioactivation and provided four additional modification patterns. Finally, 24 BPA-modified proteins were identified from the liver of BPA-exposed rats using proteomic analysis, and they were mainly enriched in oxidative stress-related pathways. The modification on superoxide dismutases, catalase, and glutathione S-transferases disrupted their enzymatic functions, leading to oxidative damage. These results revealed that the covalent protein modification is an unignorable factor for BPA hepatotoxicity. Moreover, the workflow can be applied to identify protein adducts of other emerging contaminants and possible risk.