Abstract
Apurinic/apyrimidinic endonuclease 1 (APE1) plays an important role in protecting the fidelity of genetic material and is also a promising biomarker of cancers. Herein, a simple and sensitive fluorescent method for APE1 activity detection was designed based on host-guest interaction between β-cyclodextrin polymer (β-CDP) and pyrene. In this method, pyrene-labelled DNA probes (AP-S1S2) and β-CDP functioned as fluorescent signal producer and enhancer, respectively. When APE1 was absent, pyrene on the AP-S1S2 could not enter the cavity of β-CDP because of steric hindrance, leading to a weak fluorescence intensity/anisotropy. When APE1 was present, it would recognize and cleave AP site in AP-S1S2, thus producing a dissociated pyrene-labelled oligonucleotide segment. Pyrene labelled at the 5′ end of oligonucleotide segment could be easily trapped into the cavity of β-CDP via host-guest interaction, leading to a significant enhancement of fluorescence intensity/anisotropy. This assay offered a dynamic range of 0.05U/mL–5U/mL and an estimated detection limit of 0.05U/mL. Furthermore, assessment of APE1 activity in HeLa cell extracts was successfully implemented, which hold the potential for APE1-related cancer diagnosis and biological researches.
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