2024-03-19T13:18:28Zhttp://digital.csic.es/dspace-oai/requestoai:digital.csic.es:10261/1298192020-05-28T13:37:31Zcom_10261_5063com_10261_5col_10261_5066
00925njm 22002777a 4500
dc
Tintoré, María
author
Eritja Casadellà, Ramón
author
Grijalvo, Santiago
author
Fàbrega, Carme
author
2015-11-15
O6-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.
Bioorganic and Medicinal Chemistry Letters
http://hdl.handle.net/10261/129819
10.1016/j.bmcl.2015.09.065
http://dx.doi.org/10.13039/501100003329
DNA repair
Fluorescence
Mitsunobu
O6-alkylguanine-DNA alkyltransferase
Oligonucleotide synthesis
Synthesis of oligonucleotides carrying fluorescently labelled O6-alkylguanine for measuring hAGT activity