RT Journal Article
T1 Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic.
A1 Ariza, Adriana
A1 Collado, Daniel
A1 Vida, Yolanda
A1 Montañez, María I
A1 Pérez-Inestrosa, Ezequiel
A1 Blanca, Miguel
A1 Torres, María José
A1 Cañada, F Javier
A1 Pérez-Sala, Dolores
K1 Animales
K1 Antibacterianos
K1 Biotinilación
K1 Butilaminas
K1 Haptenos
K1 Hipersensibilidad
K1 Amoxicilina
K1 Inmunoglobulina E
K1 Macrófagos
K1 Estructura molecular
K1 Beta-lactamas
AB 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.
PB Public Library of Science
YR 2014
FD 2014-03-03
LK http://hdl.handle.net/10668/1985
UL http://hdl.handle.net/10668/1985
LA en
NO Ariza A, Collado D, Vida Y, Montañez MI, Pérez-Inestrosa E, Blanca M, et al. Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic. PLoS ONE. 2014; 9(3):e90891
NO Journal Article; Research Support, Non-U.S. Gov't;
DS RISalud
RD Apr 6, 2025