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HR-MAS NMR Spectroscopy of Reconstructed Human Epidermis: Potential for the in situ Investigation of the Chemical Interactions between Skin Allergens and Nucleophilic Amino Acids.
MedLine Citation:
PMID:  23256819     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
High-resolution magic angle spinning (HR-MAS) is a nuclear magnetic resonance (NMR) technique that enables the characterization of metabolic phenotypes/metabolite profiles of cells, tissues and organs, under both normal and pathological conditions, without resorting to time-consuming extraction techniques. In this paper we explore a new domain of application of HR-MAS namely Reconstructed Human Epidermis (RHE) and the in situ observation of chemical interactions between skin sensitizers and nucleophilic amino acids. First, the preparation, storage and analysis of RHE were optimized and this work demonstrated that HR-MAS NMR was well adapted for investigating RHE with spectra of good quality allowing qualitative as well as quantitative studies of metabolites. Second, in order to study the response of RHE to chemical sensitizers, the (13C)methyldodecanesulfonate was chosen as an NMR probe and we compared adducts formed on Human Serum Albumin (HSA) in solution and adducts formed in RHE. Thus, while the modification of proteins or peptides in solution takes several days to lead to a significant amount of modification, in RHE the modifications of nucleophilic amino acids were observable already at 24h. The chemioselectivity also appeared to be different with major modifications taking place on histidine, methionine and cysteine residues in RHE while on HSA, significant modifications were observed on lysine residues with the formation of methylated and dimethylated amino groups. We thus demonstrated that RHE could be used to investigate in situ chemical interactions taking place between skin sensitizers and nucleophilic amino acids. This opens perspectives for the molecular understanding of the skin immune system activation by sensitizing chemicals. 
Authors:
Karim Elbayed; Valerie Berl; Camille Debeuckelaere; Francois-Marie Moussallieh; Martial Piotto; Izzie-Jacques Namer; Jean-Pierre Lepoittevin
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2012-12-20
Journal Detail:
Title:  Chemical research in toxicology     Volume:  -     ISSN:  1520-5010     ISO Abbreviation:  Chem. Res. Toxicol.     Publication Date:  2012 Dec 
Date Detail:
Created Date:  2012-12-21     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  8807448     Medline TA:  Chem Res Toxicol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
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