Synthesis of oligonucleotides carrying fluorescently labelled O(6)-alkylguanine for measuring hAGT activity.

Authors :: Tintoré M
Grijalvo S
Eritja R
Fàbrega C
Source :: Bioorganic & medicinal chemistry letters [Bioorg Med Chem Lett] 2015 Nov 15; Vol. 25 (22), pp. 5208-11. Date of Electronic Publication: 2015 Sep 30.
Publication Year :: 2015
Abstract: O(6)-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 O(6)-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.<br /> (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Subjects :: Base Pair Mismatch
Fluoresceins chemistry
Fluorescent Dyes chemistry
Humans
Oligonucleotides chemistry
Enzyme Assays methods
Fluorescent Dyes chemical synthesis
O(6)-Methylguanine-DNA Methyltransferase analysis
Oligonucleotides chemical synthesis

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