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Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation
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, i...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8793629/ https://www.ncbi.nlm.nih.gov/pubmed/35095517 http://dx.doi.org/10.3389/fphar.2021.807742 |
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author | González-Morena, Juan M. Sánchez-Gómez, Francisco J. Vida, Yolanda Pérez-Inestrosa, Ezequiel Salas, María Montañez, María I. Altomare, Alessandra Aldini, Giancarlo Pajares, María A. Pérez-Sala, Dolores |
author_facet | González-Morena, Juan M. Sánchez-Gómez, Francisco J. Vida, Yolanda Pérez-Inestrosa, Ezequiel Salas, María Montañez, María I. Altomare, Alessandra Aldini, Giancarlo Pajares, María A. Pérez-Sala, Dolores |
author_sort | González-Morena, Juan M. |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-8793629 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-87936292022-01-28 Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation González-Morena, Juan M. Sánchez-Gómez, Francisco J. Vida, Yolanda Pérez-Inestrosa, Ezequiel Salas, María Montañez, María I. Altomare, Alessandra Aldini, Giancarlo Pajares, María A. Pérez-Sala, Dolores Front Pharmacol Pharmacology 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. Frontiers Media S.A. 2022-01-13 /pmc/articles/PMC8793629/ /pubmed/35095517 http://dx.doi.org/10.3389/fphar.2021.807742 Text en Copyright © 2022 González-Morena, Sánchez-Gómez, Vida, Pérez-Inestrosa, Salas, Montañez, Altomare, Aldini, Pajares and Pérez-Sala. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Pharmacology González-Morena, Juan M. Sánchez-Gómez, Francisco J. Vida, Yolanda Pérez-Inestrosa, Ezequiel Salas, María Montañez, María I. Altomare, Alessandra Aldini, Giancarlo Pajares, María A. Pérez-Sala, Dolores Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation |
title | Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation |
title_full | Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation |
title_fullStr | Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation |
title_full_unstemmed | Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation |
title_short | Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation |
title_sort | amoxicillin haptenation of α-enolase is modulated by active site occupancy and acetylation |
topic | Pharmacology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8793629/ https://www.ncbi.nlm.nih.gov/pubmed/35095517 http://dx.doi.org/10.3389/fphar.2021.807742 |
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