Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic.
Allergic reactions towards β-lactam antibiotics pose an important clinical problem. The ability of small molecules, such as a β-lactams, to bind covalently to proteins, in a process known as haptenation, is considered necessary for induction of a specific immunological response. Identification of th...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2014-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3940954?pdf=render |
| _version_ | 1811270424585043968 |
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| author | Adriana Ariza Daniel Collado Yolanda Vida María I Montañez Ezequiel Pérez-Inestrosa Miguel Blanca María José Torres F Javier Cañada Dolores Pérez-Sala |
| author_facet | Adriana Ariza Daniel Collado Yolanda Vida María I Montañez Ezequiel Pérez-Inestrosa Miguel Blanca María José Torres F Javier Cañada Dolores Pérez-Sala |
| author_sort | Adriana Ariza |
| collection | DOAJ |
| description | Allergic reactions towards β-lactam antibiotics pose an important clinical problem. The ability of small molecules, such as a β-lactams, to bind covalently to proteins, in a process known as haptenation, is considered necessary for induction of a specific immunological response. Identification of the proteins modified by β-lactams and elucidation of the relevance of this process in allergic reactions requires sensitive tools. Here we describe the preparation and characterization of a biotinylated amoxicillin analog (AX-B) as a tool for the study of protein haptenation by amoxicillin (AX). AX-B, obtained by the inclusion of a biotin moiety at the lateral chain of AX, showed a chemical reactivity identical to AX. Covalent modification of proteins by AX-B was reduced by excess AX and vice versa, suggesting competition for binding to the same targets. From an immunological point of view, AX and AX-B behaved similarly in RAST inhibition studies with sera of patients with non-selective allergy towards β-lactams, whereas, as expected, competition by AX-B was poorer with sera of AX-selective patients, which recognize AX lateral chain. Use of AX-B followed by biotin detection allowed the observation of human serum albumin (HSA) modification by concentrations 100-fold lower that when using AX followed by immunological detection. Incubation of human serum with AX-B led to the haptenation of all of the previously identified major AX targets. In addition, some new targets could be detected. Interestingly, AX-B allowed the detection of intracellular protein adducts, which showed a cell type-specific pattern. This opens the possibility of following the formation and fate of AX-B adducts in cells. Thus, AX-B may constitute a valuable tool for the identification of AX targets with high sensitivity as well as for the elucidation of the mechanisms involved in allergy towards β-lactams. |
| first_indexed | 2024-04-12T22:01:43Z |
| format | Article |
| id | doaj.art-6e7615c1f0a34aa0b2f1b10e9e5d2d91 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-12T22:01:43Z |
| publishDate | 2014-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-6e7615c1f0a34aa0b2f1b10e9e5d2d912022-12-22T03:15:05ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0193e9089110.1371/journal.pone.0090891Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic.Adriana ArizaDaniel ColladoYolanda VidaMaría I MontañezEzequiel Pérez-InestrosaMiguel BlancaMaría José TorresF Javier CañadaDolores Pérez-SalaAllergic reactions towards β-lactam antibiotics pose an important clinical problem. The ability of small molecules, such as a β-lactams, to bind covalently to proteins, in a process known as haptenation, is considered necessary for induction of a specific immunological response. Identification of the proteins modified by β-lactams and elucidation of the relevance of this process in allergic reactions requires sensitive tools. Here we describe the preparation and characterization of a biotinylated amoxicillin analog (AX-B) as a tool for the study of protein haptenation by amoxicillin (AX). AX-B, obtained by the inclusion of a biotin moiety at the lateral chain of AX, showed a chemical reactivity identical to AX. Covalent modification of proteins by AX-B was reduced by excess AX and vice versa, suggesting competition for binding to the same targets. From an immunological point of view, AX and AX-B behaved similarly in RAST inhibition studies with sera of patients with non-selective allergy towards β-lactams, whereas, as expected, competition by AX-B was poorer with sera of AX-selective patients, which recognize AX lateral chain. Use of AX-B followed by biotin detection allowed the observation of human serum albumin (HSA) modification by concentrations 100-fold lower that when using AX followed by immunological detection. Incubation of human serum with AX-B led to the haptenation of all of the previously identified major AX targets. In addition, some new targets could be detected. Interestingly, AX-B allowed the detection of intracellular protein adducts, which showed a cell type-specific pattern. This opens the possibility of following the formation and fate of AX-B adducts in cells. Thus, AX-B may constitute a valuable tool for the identification of AX targets with high sensitivity as well as for the elucidation of the mechanisms involved in allergy towards β-lactams.http://europepmc.org/articles/PMC3940954?pdf=render |
| spellingShingle | Adriana Ariza Daniel Collado Yolanda Vida María I Montañez Ezequiel Pérez-Inestrosa Miguel Blanca María José Torres F Javier Cañada Dolores Pérez-Sala Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic. PLoS ONE |
| title | Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic. |
| title_full | Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic. |
| title_fullStr | Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic. |
| title_full_unstemmed | Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic. |
| title_short | Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic. |
| title_sort | study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic |
| url | http://europepmc.org/articles/PMC3940954?pdf=render |
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