کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1226620 1494803 2012 17 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Protein haptenation by amoxicillin: High resolution mass spectrometry analysis and identification of target proteins in serum
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Protein haptenation by amoxicillin: High resolution mass spectrometry analysis and identification of target proteins in serum
چکیده انگلیسی

Allergy towards wide spectrum antibiotics such as amoxicillin (AX) is a major health problem. Protein haptenation by covalent conjugation of AX is considered a key process for the allergic response. However, the nature of the proteins involved has not been completely elucidated. Human serum albumin (HSA) is the most abundant protein in plasma and is considered a major target for haptenation by drugs, including β-lactam antibiotics. Here we report a procedure for immunological detection of AX–protein adducts with antibodies recognizing the lateral chain of the AX molecule. With this approach we detected human serum proteins modified by AX in vitro and identified HSA, transferrin and immunoglobulins heavy and light chains as prominent AX-modified proteins. Since HSA was the major AX target, we characterized AX–HSA interaction using high resolution LTQ orbitrap MS. At 0.5 mg/mL AX, we detected one main AX–HSA adduct involving residues Lys 190, 199 or 541, whereas higher AX concentrations elicited a more extensive modification. In molecular modeling studies Lys190 and Lys 199 were found the most reactive residues towards AX, with surrounding residues favoring adduct formation. These findings provide novel tools and insight for the study of protein haptenation and the mechanisms involved in AX-elicited allergic reactions.

Figure optionsDownload high-quality image (186 K)Download as PowerPoint slideHighlights
► Development of a method for immunological detection of amoxicillin-modified proteins
► Amoxicillin binds selectively to several proteins in human serum.
► High resolution MS analysis of amoxicillin-modified human serum albumin
► Amoxicillin at low concentrations binds to Lys190, Lys199 or 541 in human serum albumin.
► Importance of elucidation of protein haptenation by amoxicillin in drug allergy study

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Proteomics - Volume 77, 21 December 2012, Pages 504–520
نویسندگان
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