Citation: Zhou Maogui, Qin Shiya, Feng Zhenzhen, Song Chunxia, Zhang Hua, Li Wenshan, Wang Qing, Liu Jianbo, Huang Jin, Yang Xiaohai, Wang Kemin. A simple and sensitive assay for apurinic/apyrimidinic endonuclease 1 activity based on host-guest interaction of β-cyclodextrin polymer and pyrene[J]. Chinese Chemical Letters, ;2018, 29(6): 973-976. doi: 10.1016/j.cclet.2017.10.010 shu

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

Zhou Maogui ^a,1 ,
Qin Shiya ^a,1 ,
Feng Zhenzhen ^a ,
Song Chunxia ^b ,
Zhang Hua ^a ,
Li Wenshan ^a ,
Wang Qing ^a ,
Liu Jianbo ^a ,
Huang Jin ^a ,
Yang Xiaohai ^a,, ,
Wang Kemin ^a,,

a.
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, Changsha 410082, China
b.
School of Science, Anhui Agricultural University, Hefei 230036, China

Corresponding author: Yang Xiaohai, yangxiaohai@hnu.edu.cn Wang Kemin, kmwang@hnu.edu.cn

Received Date: 31 May 2017
Revised Date: 9 October 2017
Accepted Date: 11 October 2017
Available Online: 13 June 2017

Figures(5)

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 APS1S2, 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 0.05 U/mL-5 U/mL and an estimated detection limit of 0.05 U/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.
- APE1,
- Host-guest interaction,
- Fluorescence,
- β-Cyclodextrin polymer,
- Pyrene
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