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...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-01-01
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| Series: | Frontiers in Pharmacology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fphar.2021.807742/full |
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| author | Juan M. González-Morena Francisco J. Sánchez-Gómez Yolanda Vida Yolanda Vida Ezequiel Pérez-Inestrosa Ezequiel Pérez-Inestrosa María Salas María I. Montañez Alessandra Altomare Giancarlo Aldini María A. Pajares Dolores Pérez-Sala |
| author_facet | Juan M. González-Morena Francisco J. Sánchez-Gómez Yolanda Vida Yolanda Vida Ezequiel Pérez-Inestrosa Ezequiel Pérez-Inestrosa María Salas María I. Montañez Alessandra Altomare Giancarlo Aldini María A. Pajares Dolores Pérez-Sala |
| author_sort | Juan M. González-Morena |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-20T07:09:25Z |
| format | Article |
| id | doaj.art-dca58d595c514fb59582cecf556410c6 |
| institution | Directory Open Access Journal |
| issn | 1663-9812 |
| language | English |
| last_indexed | 2024-12-20T07:09:25Z |
| publishDate | 2022-01-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Pharmacology |
| spelling | doaj.art-dca58d595c514fb59582cecf556410c62022-12-21T19:48:55ZengFrontiers Media S.A.Frontiers in Pharmacology1663-98122022-01-011210.3389/fphar.2021.807742807742Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and AcetylationJuan M. González-Morena0Francisco J. Sánchez-Gómez1Yolanda Vida2Yolanda Vida3Ezequiel Pérez-Inestrosa4Ezequiel Pérez-Inestrosa5María Salas6María I. Montañez7Alessandra Altomare8Giancarlo Aldini9María A. Pajares10Dolores Pérez-Sala11Department of Structural and Chemical Biology, Centro de Investigaciones Biológicas Margarita Salas (CSIC), Madrid, SpainDepartment of Structural and Chemical Biology, Centro de Investigaciones Biológicas Margarita Salas (CSIC), Madrid, SpainDpto. Química Orgánica, Universidad de Málaga-IBIMA, Málaga, SpainCentro Andaluz de Nanomedicina y Biotecnología-BIONAND, Parque Tecnológico de Andalucía, Málaga, SpainDpto. Química Orgánica, Universidad de Málaga-IBIMA, Málaga, SpainCentro Andaluz de Nanomedicina y Biotecnología-BIONAND, Parque Tecnológico de Andalucía, Málaga, SpainAllergy Research Group, Instituto de Investigación Biomédica de Málaga-IBIMA, Allergy Unit, Hospital Regional Universitario de Málaga, Málaga, SpainAllergy Research Group, Instituto de Investigación Biomédica de Málaga-IBIMA, Andalusian Centre for Nanomedicine Biotechnology-BIONAND, Parque Tecnológico de Andalucía, Málaga, SpainDepartment of Scienze Farmaceutiche, Universita degli Studi di Milano, Milan, ItalyDepartment of Scienze Farmaceutiche, Universita degli Studi di Milano, Milan, ItalyDepartment of Structural and Chemical Biology, Centro de Investigaciones Biológicas Margarita Salas (CSIC), Madrid, SpainDepartment of Structural and Chemical Biology, Centro de Investigaciones Biológicas Margarita Salas (CSIC), Madrid, SpainAllergic 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.https://www.frontiersin.org/articles/10.3389/fphar.2021.807742/fullbeta-lactam antibioticsprotein modification by drugsmass spectrometryposttranslational modificationacetylationallergic responses to drugs |
| spellingShingle | Juan M. González-Morena Francisco J. Sánchez-Gómez Yolanda Vida Yolanda Vida Ezequiel Pérez-Inestrosa Ezequiel Pérez-Inestrosa María Salas María I. Montañez Alessandra Altomare Giancarlo Aldini María A. Pajares Dolores Pérez-Sala Amoxicillin Haptenation of α-Enolase is Modulated by Active Site Occupancy and Acetylation Frontiers in Pharmacology beta-lactam antibiotics protein modification by drugs mass spectrometry posttranslational modification acetylation allergic responses to drugs |
| 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 | beta-lactam antibiotics protein modification by drugs mass spectrometry posttranslational modification acetylation allergic responses to drugs |
| url | https://www.frontiersin.org/articles/10.3389/fphar.2021.807742/full |
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