Label-Free Sensing of Human 8-Oxoguanine DNA Glycosylase Activity with a Nanopore

被引：37

作者：

Shang, Jizhen ^{[1
,2
]}

Li, Zhi ^{[1
]}

Liu, Liping ^{[1
]}

Xi, Dongmei ^{[1
]}

Wang, Hua ^{[2
]}

机构：

[1] Linyi Univ, Coll Chem & Chem Engn, Shandong Prov Key Lab Detect Technol Tumor Makers, Linyi 276005, Peoples R China

[2] Qufu Normal Univ, Coll Chem & Chem Engn, Inst Med & Mat Applicat Technol, Qufu 273165, Peoples R China

来源：

ACS SENSORS | 2018年 / 3卷 / 02期

基金：

中国国家自然科学基金;

关键词：

nanopore sensor; hOGG1; enzyme-catalytic cleavage reaction; DNA substrate; 8-oxoguanine; BASE-EXCISION-REPAIR; LUNG-CANCER; METAL-IONS; AMPLIFICATION; MICRORNAS; DAMAGE; ASSAY; PROBE; RISK; PROTEIN;

D O I：

10.1021/acssensors.7b00954

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Human 8-oxoguanine DNA glycosylase (hOGG1) plays a significant role in maintaining the genomic integrity of living organisms for its capability of repairing DNA lesions. Accurate detection of hOGG1 activity would greatly facilitate the screening and early diagnosis of diseases. In this work, we report a nanopore-based sensing strategy to probe the hOGG1 activity by employing the enzyme-catalytic cleavage reaction of DNA substrate. The hOGG1 specifically catalyzed the removal of the 8-hydroxyguanine (8-oxoG) and cleaved the DNA substrates immobilized on magnetic beads, thereby releasing the output DNA which would quantitatively produce the signature current events when subjected to alpha-hemolysin (alpha-HL) nanopore test. The approach enables the sensitive detection of hOGG1 activity without the need of any labeling or signal amplification route. Furthermore, the method can be applied to assay the inhibition of hOGG1 and evaluate the activity of endogenous hOGG1 in crude cell extracts. Importantly, since DNAs with specific sequences are the catalytic substrates of a wide variety of enzymes, the proposed strategy should be universally applicable for probing the activities of different types of enzymes with nanopore sensors.

引用

页码：512 / 518

页数：13