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Synthesis of oligonucleotides carrying fluorescently labelled O6-alkylguanine for measuring hAGT activity

AuthorsTintoré, María ; Eritja Casadellà, Ramón ; Grijalvo, Santiago ; Fàbrega, Carme
KeywordsDNA repair
O6-alkylguanine-DNA alkyltransferase
Oligonucleotide synthesis
Issue Date15-Nov-2015
CitationBioorganic and Medicinal Chemistry Letters
AbstractO6-alkylguanine-DNA-alkyltransferase (hAGT) activity provides resistance to cancer chemotherapeutic agents and its inhibition enhances chemotherapy. We herein present the development of a novel fluorescence assay for the detection of hAGT activity. We designed a dsDNA sequence containing a fluorophore-quencher pair, where the fluorophore was attached to an O6-benzylguanine. This precursor was synthesized using the Mitsunobu reaction to introduce the benzyl group. The alkyl-fluorophore group is transferred to the active site during the dealkylation, producing an increase in fluorescence which is correlated to hAGT activity. This assay can be used for the evaluation of potential inhibitors of hAGT in a straightforward manner.
Publisher version (URL)http://www.sciencedirect.com/science/article/pii/S0960894X15300998
Appears in Collections:(IQAC) Artículos
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