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

AutorTintoré, María; Eritja Casadellà, Ramón; Grijalvo, Santiago; Fàbrega, Carme
Palabras claveDNA repair
Fluorescence
Mitsunobu
O6-alkylguanine-DNA alkyltransferase
Oligonucleotide synthesis
Fecha de publicación15-nov-2015
EditorElsevier
CitaciónBioorganic and Medicinal Chemistry Letters
ResumenO6-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.
Versión del editorhttp://www.sciencedirect.com/science/article/pii/S0960894X15300998
URIhttp://hdl.handle.net/10261/129819
DOI10.1016/j.bmcl.2015.09.065
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