2026-05-17T01:44:09Zhttps://repisalud.isciii.es/rest/oai/request

oai:repisalud.isciii.es:20.500.12105/185782024-02-27T14:57:48Zcom_20.500.12105_15322com_20.500.12105_2051col_20.500.12105_16927

00925njm 22002777a 4500 dc González-Morena, Juan M author Sánchez-Gómez, Francisco J author Vida, Yolanda author Pérez-Inestrosa, Ezequiel author Salas, María author Montañez, María I author Altomare, Alessandra author Aldini, Giancarlo author Pajares, María A author Pérez-Sala, Dolores author 2022-01-13 Allergic reactions to antibiotics are a major concern in the clinic. ß-lactam antibiotics are the class most frequently reported to cause hypersensitivity reactions. One of the mechanisms involved in this outcome is the modification of proteins by covalent binding of the drug (haptenation). Hence, interest in identifying the corresponding serum and cellular protein targets arises. Importantly, haptenation susceptibility and extent can be modulated by the context, including factors affecting protein conformation or the occurrence of other posttranslational modifications. We previously identified the glycolytic enzyme α-enolase as a target for haptenation by amoxicillin, both in cells and in the extracellular milieu. Here, we performed an in vitro study to analyze amoxicillin haptenation of α-enolase using gel-based and activity assays. Moreover, the possible interplay or interference between amoxicillin haptenation and acetylation of α-enolase was studied in 1D- and 2D-gels that showed decreased haptenation and displacement of the haptenation signal to lower pI spots after chemical acetylation of the protein, respectively. In addition, the peptide containing lysine 239 was identified by mass spectrometry as the amoxicillin target sequence on α-enolase, thus suggesting a selective haptenation under our conditions. The putative amoxicillin binding site and the surrounding interactions were investigated using the α-enolase crystal structure and molecular docking. Altogether, the results obtained provide the basis for the design of novel diagnostic tools or approaches in the study of amoxicillin-induced allergic reactions. 10.3389/fphar.2021.807742 1663-9812 Frontiers in pharmacology http://hdl.handle.net/10668/20700 35095517 http://hdl.handle.net/20.500.12105/18578 acetylation allergic responses to drugs beta-lactam antibiotics mass spectrometry posttranslational modification protein modification by drugs Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation.