کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
2581110 | 1561639 | 2012 | 10 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Interactions of organophosphates with keratins in the cornified epithelium of human skin Interactions of organophosphates with keratins in the cornified epithelium of human skin](/preview/png/2581110.png)
Methods to unequivocally assess and quantify exposure to organophosphate anti-cholinesterase agents are highly valuable, either from a biomonitoring or a forensic perspective. Since for both OP pesticides and various nerve agents the skin is a predominant route of entry, we hypothesized that proteins in the skin might represent an ideal source of unequivocal and persistent biomarkers for exposure to these compounds. In this exploratory study we show that keratin proteins in human skin are relevant binding sites for organophosphates. The thick cornified epithelium of human plantar skin (callus) was exposed to a selection of relevant organophosphorus compounds and keratin proteins were subsequently extracted. After carboxymethylation of cysteine residues, enzymatic digestion of the keratins with pronase and trypsin was performed and the resulting amino acid and peptides were analyzed to assess whether covalent adducts had formed. LC-tandem MS analysis of the pronase digests demonstrated that tyrosine and to a lesser extent serine residues were selectively modified by organophosphate pesticides (both phosphorothioates and the corresponding oxon forms) under physiological conditions. In addition, modification of tyrosine with the nerve agent VX was unequivocally assessed. In order to elucidate specific binding sites, LC-tandem MS analysis of trypsin digests showed two separate tryptic keratin fragments, i.e. LASY*LDK and SLY*GLGGSK, with Y* the modified tyrosine residues, originating from keratin 1/6 and keratin 10, respectively. These preliminary findings, revealing novel binding targets for anti-cholinesterase organophosphates, will form a firm basis for the development of novel (non-invasive) methods for assessment of exposure to organophosphates. Whether this binding will also have biological implications remains an issue for further investigations.
► We have explored alternative binding sites for organophosphates in human skin.
► Upon exposure of human callus to organophosphates, covalent protein adducts are formed.
► Phosphylated Tyr and Ser adducts were released by pronase treatment of OP-exposed keratin.
► Two tryptic keratin fragments were identified containing a phosphylated Tyr residue.
Journal: Chemico-Biological Interactions - Volume 197, Issues 2–3, 30 May 2012, Pages 93–102