Covalent Histone Modification by an Electrophilic Derivative of the Anti-HIV Drug Nevirapine

• Published in: Molecules (Basel, Switzerland) Switzerland), 2021-03, Vol.26 (5), p.1349
• Main Authors: Harjivan, Shrika G, Charneira, Catarina, Martins, Inês L, Pereira, Sofia A, Espadas, Guadalupe, Sabidó, Eduard, Beland, Frederick A, Marques, M Matilde, Antunes, Alexandra M M
• Format: Article
• Language: English
• Subjects: Anti-HIV Agents - chemistry; Anti-HIV Agents - metabolism; covalent histone modification; covalent protein adducts; drug-induced adverse reactions; Histones - chemistry; Histones - metabolism; Humans; mass spectrometry; Molecular Structure; nevirapine; Nevirapine - chemistry; Nevirapine - metabolism; Proteome - analysis
• ISSN: 1420-3049; 1420-3049
• DOI: 10.3390/molecules26051349

Nevirapine (NVP), a non-nucleoside reverse transcriptase inhibitor widely used in combined antiretroviral therapy and to prevent mother-to-child transmission of the human immunodeficiency virus type 1, is associated with several adverse side effects. Using 12-mesyloxy-nevirapine, a model electrophile of the reactive metabolites derived from the NVP Phase I metabolite, 12-hydroxy-NVP, we demonstrate that the nucleophilic core and -terminal residues of histones are targets for covalent adduct formation. We identified multiple NVP-modification sites at lysine (e.g., H2BK47, H4K32), histidine (e.g., H2BH110, H4H76), and serine (e.g., H2BS33) residues of the four histones using a mass spectrometry-based bottom-up proteomic analysis. In particular, H2BK47, H2BH110, H2AH83, and H4H76 were found to be potential hot spots for NVP incorporation. Notably, a remarkable selectivity to the imidazole ring of histidine was observed, with modification by NVP detected in three out of the 11 histidine residues of histones. This suggests that NVP-modified histidine residues of histones are prospective markers of the drug's bioactivation and/or toxicity. Importantly, NVP-derived modifications were identified at sites known to determine chromatin structure (e.g., H4H76) or that can undergo multiple types of post-translational modifications (e.g., H2BK47, H4H76). These results open new insights into the molecular mechanisms of drug-induced adverse reactions.