2024-03-29T01:22:07Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/984202021-12-27T16:20:08Zcom_10261_86com_10261_1col_10261_339
2014-06-16T11:32:44Z
urn:hdl:10261/98420
Study of protein haptenation by amoxicillin through the use of a biotinylated antibiotic
Ariza, Adriana
Collado, Daniel
Vida, Yolanda
Montañez, M. I.
Pérez-Inestrosa, Ezequiel
Blanca, Miguel
Torres, María J.
Cañada, F. Javier
Pérez-Sala, Dolores
12 p.-9 fig.-2 tab.
Allergic reactions towards b-lactam antibiotics pose an important clinical problem. The ability of small molecules, such as a b-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 b-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 b-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 b-lactams.
2014-06-16T11:32:44Z
2014-06-16T11:32:44Z
2014-03-03
artículo
PLoS ONE 9(3): e90891
1932-6203
http://hdl.handle.net/10261/98420
10.1371/journal.pone.0090891
1932-6203
24595455
eng
Publisher's version
http://dx.doi.org/10.1371/journal.pone.0090891
Sí
FP7/2007-2013
http://creativecommons.org/licenses/by/4.0/
openAccess
Public Library of Science