Institutional Repository of Key Laboratory of Coastal Zone Environmental Processes, Yantai Institute of Coastal Zone Research, Chinese Academy of Sciences (KLCEP)
| Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode | |
Ding, Jiawang1,2,3; Qin, Wei1,2
| |
| 发表期刊 | BIOSENSORS & BIOELECTRONICS
 |
| ISSN | 0956-5663 |
| 2013-09-15 | |
| 卷号 | 47页码:559-565 |
| 关键词 | Nucleases Hydroxyl Radicals Potentiometry Protamine Polyion Sensors |
| 产权排序 | [Ding, Jiawang; Qin, Wei] Chinese Acad Sci, Key Lab Coastal Zone Environm Proc & Ecol Remedia, Yantai Inst Coastal Zone Res YIC, Yantai 264003, Shandong, Peoples R China; [Ding, Jiawang; Qin, Wei] YICCAS, Shandong Prov Key Lab Coastal Zone Environm Proc, Yantai 264003, Shandong, Peoples R China; [Ding, Jiawang] Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 通讯作者 | Qin, W (reprint author), Chinese Acad Sci, Key Lab Coastal Zone Environm Proc & Ecol Remedia, Yantai Inst Coastal Zone Res YIC, Yantai 264003, Shandong, Peoples R China. [email protected] |
| 作者部门 | 中科院海岸带环境过程与生态修复重点实验室 |
| 英文摘要 | A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7 x 10(-4) U/mu L for S1 nuclease, and of 3.9 x 10(-4) U/mu L for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2 x 10(-4) U/mu L for S1 nuclease, and of 4.5 x 10(-4) U/mu L for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. (C) 2013 Elsevier B.V. All rights reserved.; A simple, general and label-free potentiometric method to measure nuclease activities and oxidative DNA damage in a homogeneous solution using a polycation-sensitive membrane electrode is reported. Protamine, a linear polyionic species, is used as an indicator to report the cleavage of DNA by nucleases such as restriction and nonspecific nucleases, and the damage of DNA induced by hydroxyl radicals. Measurements can be done with a titration mode or a direct detection mode. For the potentiometric titration mode, the enzymatic cleavage dramatically affects the electrostatical interaction between DNA and protamine and thus shifts the response curve for the potentiometric titration of the DNA with protamine. Under the optimized conditions, the enzyme activities can be sensed potentiometrically with detection limits of 2.7 x 10(-4) U/mu L for S1 nuclease, and of 3.9 x 10(-4) U/mu L for DNase I. For the direct detection mode, a biocomplex between protamine and DNA is used as a substrate. The nuclease of interest cleaves the DNA from the protamine/DNA complex into smaller fragments, so that free protamine is generated and can be detected potentiometrically via the polycation-sensitive membrane electrode. Using a direct measurement, the nuclease activities could be rapidly detected with detection limits of 3.2 x 10(-4) U/mu L for S1 nuclease, and of 4.5 x 10(-4) U/mu L for DNase I. Moreover, the proposed potentiometric assays demonstrate the potential applications in the detection of hydroxyl radicals. It is anticipated that the present potentiometric strategy will provide a promising platform for high-throughput screening of nucleases, reactive oxygen species and the drugs with potential inhibition abilities. (C) 2013 Elsevier B.V. All rights reserved. |
| 文章类型 | Article |
| 资助机构 | Chinese Academy of Sciences [YZ201161]; National Natural Science Foundation of China [41176081, 21207156]; Science and Technology Project of Yantai [2012132]; Taishan Scholar Program of Shandong Province [TS20081159] |
| 收录类别 | SCI |
| 语种 | 英语 |
| 关键词[WOS] | LABEL-FREE ; RESTRICTION ENDONUCLEASES ; ELECTROCHEMICAL ASSAY ; GOLD NANOPARTICLES ; ENZYMATIC CLEAVAGE ; DETECTION LIMITS ; BIOSENSORS ; SYSTEM ; INHIBITION ; SUBSTRATE |
| 研究领域[WOS] | Biophysics ; Biotechnology & Applied Microbiology ; Chemistry ; Electrochemistry ; Science & Technology - Other Topics |
| WOS记录号 | WOS:000320481900085 |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.yic.ac.cn/handle/133337/6652 |
| 专题 | 中国科学院海岸带环境过程与生态修复重点实验室 |
| 作者单位 | 1.Chinese Acad Sci, Key Lab Coastal Zone Environm Proc & Ecol Remedia, Yantai Inst Coastal Zone Res YIC, Yantai 264003, Shandong, Peoples R China 2.YICCAS, Shandong Prov Key Lab Coastal Zone Environm Proc, Yantai 264003, Shandong, Peoples R China 3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Ding, Jiawang,Qin, Wei. Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode[J]. BIOSENSORS & BIOELECTRONICS,2013,47:559-565. |
| APA | Ding, Jiawang,&Qin, Wei.(2013).Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode.BIOSENSORS & BIOELECTRONICS,47,559-565. |
| MLA | Ding, Jiawang,et al."Potentiometric sensing of nuclease activities and oxidative damage of single-stranded DNA using a polycation-sensitive membrane electrode".BIOSENSORS & BIOELECTRONICS 47(2013):559-565. |
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