A simple and sensitive assay for apurinic/apyrimidinic endonuclease 1activity based on host-guest interaction of β-cyclodextrin polymer and pyrene

查看全文 作　　者：Maogui [1]Zhou;Shiya [1]Qin;Zhenzhen [1]Feng;Chunxia [2]Song;Hua [1]Zhang;Wenshan [1]Li;Qing [1]Wang;Jianbo [1]Liu;Jin [1]Huang;Xiaohai [1]Yang;Kemin [1]Wang 机构地区：[1]State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Key Laboratory for Bio-Nanotechnology and Molecular Engineering of Hunan Province, Hunan University;[2]School of Science, Anhui Agricultural University 出　　处：《Chinese Chemical Letters》索引2018年第29卷第6期,共4页 基　　金：supported by the National Natural Science Foundation of China (Nos. 21675047, 21375034, 21190040);the Foundation for Innovative Research Groups of National Natural Science Foundation of China (No. 21521063) 摘　　要：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-S1 S2) and β-CDP functioned as fluorescent signal producer and enhancer, respectively. When APE1 was absent, pyrene on the AP-S1 S2 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 APS1 S2, thus producing a dissociated pyrene-labelled oligonucleotide segment. Pyrene labelled at the 50 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 0.05 U/m L–5 U/m L and an estimated detection limit of 0.05 U/m L. Furthermore,assessment of APE1 activity in He La cell extracts was successfully implemented, which hold the potential for APE1-related cancer diagnosis and biological researches. 关 键 词：相互作用 聚合物 客人 主人 金基 敏感 荧光灯 CDP