کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1163253 | 1490936 | 2015 | 9 صفحه PDF | دانلود رایگان |
![عکس صفحه اول مقاله: Detection of cysteine- and lysine-based protein adductions by reactive metabolites of 2,5-dimethylfuran Detection of cysteine- and lysine-based protein adductions by reactive metabolites of 2,5-dimethylfuran](/preview/png/1163253.png)
• A selective, effective and convenient platform was developed to analyze protein adductions by reactive metabolites of furans.
• The technique can differentiate between cysteine- and lysine-based protein adductions by the reactive metabolites.
• The bromine-tagged reagents applied act not only as trapping agents but also as labeling agents.
Many furan-containing compounds are known to be toxic and/or carcinogenic. Metabolic activation of toxic furans to cis-enediones (cis-enedials or γ-ketoenals) is generally considered as the initial step towards the processes of their toxicities. Sequential modification of key proteins by the electrophilic reactive intermediates is suggested to be an important mechanism of the toxic actions. In the present study, we developed a novel and simple analytical platform to detect protein modification resulting from metabolic activation of model compound 2,5-dimethylfuran (DMF). 4-Bromobenzylamine and 4-bromobenzylmercaptan were employed to trap protein adductions at cysteine and lysine residues, respectively. The resulting protein samples were proteolytically digested by chymotrypsin and Pronase E, followed by LC–MS/MS analysis. Modifications of cysteine and lysine residues of proteins were observed in microsomal incubations and animals after exposure to DMF. In conclusion, the approach established has been proven highly selective and reliable. This advance allows us not only to detect the protein adductions but also to define the structural identities of amino acid residues modified. This technique provides a unique platform to assess protein modifications arising from metabolic activation of potentially harmful furan-containing compounds. Hepatic protein adductions were found to be proportional to the hepatotoxicity of DMF.
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Journal: Analytica Chimica Acta - Volume 896, 8 October 2015, Pages 93–101