2026-06-14T03:37:32Zhttps://repisalud.isciii.es/rest/oai/requestoai:repisalud.isciii.es:20.500.12105/170602024-11-28T15:35:34Zcom_20.500.12105_15322com_20.500.12105_2051col_20.500.12105_16927
Repisalud
author
Ariza, Adriana
author
Collado, Daniel
author
Vida, Yolanda
author
Montañez, María I
author
Pérez-Inestrosa, Ezequiel
author
Blanca, Miguel
author
Torres, María José
author
Cañada, F Javier
author
Pérez-Sala, Dolores
authoraffiliation
[Ariza,A; Cañada,FJ; Pérez-Sala,D] Department of Chemical and Physical Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain. [Ariza,A; Torres,MJ] Research Laboratory Carlos Haya Hospital-IBIMA, Málaga, Spain. [Collado,D; VidaY; Pérez-Inestrosa,E] Department of Organic Chemistry, University of Málaga, Malaga, Spain. [Collado,D; Vida,Y; Montañez,MI; Pérez-Inestrosa,E] BIONAND-Andalusian Centre for Nanomedicine and Biotechnology, Parque Tecnológico de Andalucía, Málaga, Spain. [Blanca,M] Allergy Service, Hospital Carlos Haya, Málaga, Spain.
2024-01-15T18:17:08Z
2024-01-15T18:17:08Z
2014-03-03
10.1371/journal.pone.0090891
1932-6203
PloS ONE
http://hdl.handle.net/10668/1985
24595455
http://hdl.handle.net/20.500.12105/17060
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.
eng
Animales
Antibacterianos
Biotinilación
Butilaminas
Haptenos
Hipersensibilidad
Amoxicilina
Inmunoglobulina E
Macrófagos
Estructura molecular
Beta-lactamas
Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic.
research article