Identification of quinol-thioether adducted proteins excreted in the urine after I.V. administration of 2-(glutathion-S-yl)HQ in Long-Evans rats

TOXI 76

Matthew T. Labenski, labenski@pharmacy.arizona.edu, Ashley A. Fisher, fisher@pharmacy.arizona.edu, George Tsaprailis, Terrence Monks, and Serrine S. Lau, lau@pharmacy.arizona.edu. Department of Pharmacology and Toxicology, University of Arizona College of Pharmacy, 1703 E. Mabel Street, Tucson, AZ 85721
Hydroquinone (HQ) and its glutathione conjugates (GSHQs) cause renal cell necrosis by redox-cycling and adducting macromolecules, specifically in the S3 segment of the renal proximal tubules. Recent technological advancements allow for the detection of chemical-induced post-translational modifications (PTMs) that may alter cell signaling pathways or alter the structure and function of the modified proteins. We have identified such modified proteins following administration of 2-(glutathion-S-yl)HQ (GSHQ) (400µmol/kg, iv, 2hr) to Long-Evans rats. Four proteins were adducted by 2-(Cystein-S-yl)HQ (CSHQ), and 3 proteins adducted by HQ. The HQ adduct originates as a quinol-thioether adduct and then the thioether bond hydrolyzes during the protein digestion leaving just the HQ adducted. The instability of the thioether bond may explain why no C adducts are observed. The adducts are observed on K, R, and E. The site-specific identification of covalently adducted proteins is a prerequisite for understanding the biological significance of chemical-induced PTMs and the subsequent toxicological response.