Activity-based DNA-gold nanoparticle probe as colorimetric biosensor for DNA methyltransferase/glycosylase assay.
Activity-based DNA-gold nanoparticle probe as colorimetric biosensor for DNA methyltransferase/glycosylase assay.
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DOI：
10.1021/ac303575f
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发表时间：
2013-04
影响因子：
7.4
通讯作者：
Zhan Wu;Zhenkun Wu;Hao Tang;Li-Juan Tang;Jian-hui Jiang
中科院分区：
文献类型：
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作者：
Zhan Wu;Zhenkun Wu;Hao Tang;Li-Juan Tang;Jian-hui Jiang
我们开发了一种新型的生物传感器平台，用于活性DNA甲基转移酶/糖基化酶的比色检测，该平台基于DNA-金纳米颗粒（AuNP）探针的末端保护，通过机制共价捕获靶酶。这种生物传感器依靠基于活性的DNA探针对靶酶的共价捕获，从而为连接在AuNPs上的DNA探针提供终端保护，使其免受Exo I和III的降解。该传感器采用均质分析格式，表面等离子体吸收强，具有检测灵敏、快速、方便等优点。由于基于活性的探针（ABPs）对目标酶具有机械特异性，因此该策略还提供了更高的选择性，并且可以获得有关酶的丰度和活性的信息。我们利用人类DNA（胞嘧啶-5）甲基转移酶（Dnmt 1）和人类8-氧鸟嘌呤糖基化酶（hOGG 1）证明了这一策略。结果表明，随着酶活性的增加，比色响应动态增加，这意味着该策略在DNA甲基转移酶/糖基化酶检测、分子诊断和药物筛选方面具有很大的潜力。我们的策略也可以作为一种有前途和方便的方法，用于DNA修饰酶的ABPs的可视化筛选。
We have developed a novel biosensor platform for colorimetric detection of active DNA methyltransferase/glycosylase based on terminal protection of the DNA-gold nanoparticle (AuNP) probes by mechanistically covalent trapping of target enzymes. This biosensor relied on covalent capture of target enzymes by activity-based DNA probes which created terminal protection of the DNA probes tethered on AuNPs from degradation by Exo I and III. This biosensor has the advantages of having highly sensitive, rapid, and convenient detection due to its use of the homogeneous assay format and strong surface plasmon absorption. Because the activity-based probes (ABPs) are mechanistically specific to target enzymes, this strategy also offers improved selectivity and can achieve the information about both abundance and activity of the enzymes. We have demonstrated this strategy using a human DNA (cytosine-5) methyltransferase (Dnmt 1) and a human 8-oxoguanine glycosylase (hOGG 1). The results reveal that the colorimetric response increases dynamically with increasing activity of the enzymes, implying a great potential of this strategy for DNA methyltransferase/glycosylase detection and molecular diagnostics and drug screening. Our strategy can also be used as a promising and convenient approach for visualized screening of ABPs for DNA modifying enzymes.