Engineering oligonucleotide-based peroxidase mimetics for the colorimetric assay of S1 nuclease†,Analytical Methods

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Engineering oligonucleotide-based peroxidase mimetics for the colorimetric assay of S1 nuclease†
Analytical Methods ( IF 2.7 ) Pub Date : 2018-02-13 00:00:00 , DOI: 10.1039/c7ay02830j
Chuan He _{1,

2,

3,

4,

5} , Jinli Zhang _{1,

2,

3,

4,

5} , Wei Li _{1,

2,

3,

4,

5} , Yan Fu _{1,

2,

3,

4,

5}

Affiliation

Herein we propose a rational design to construct peroxidase mimetics in response to S1 nuclease by employing guanine-rich oligonucleotides and Cu²⁺ ions. The enzymatic activities of DNA–Cu(II) are highly dependent upon the binding stoichiometry between Cu²⁺ and DNA. In the H₂O₂-mediated oxidation of 3,3′,5,5′-tetramethylbenzidine, the catalytic activity of Cu²⁺ can be accelerated approximately 88 times in the presence of AG₃(T₂AG₃)₃, whereas it shows slight enhancement in the presence of a mononucleotide (GMP). The coordination of Cu²⁺ with DNA structures greatly strengthens its ability to generate reactive oxygen species. The most intriguing finding in this study is that DNA–Cu(II) complexes lose their enzymatic activities after the cleavage of DNA scaffolds by S1 nuclease. A colorimetric method was established for selectively evaluating the S1 nuclease activity with a limit of detection of 4.7 × 10⁻³ U mL⁻¹, only by using label-free oligonucleotides and copper ions without complicated synthesis and apparatus. This facile method can also be applicable for quantitative analysis in biological fluids.

中文翻译：

工程寡核苷酸基于过氧化物酶模拟物S1核酸酶的比色测定†

在本文中，我们提出了一种合理的设计，以通过使用富含鸟嘌呤的寡核苷酸和Cu ²⁺离子来构建响应S1核酸酶的过氧化物酶模拟物。DNA–Cu（II）的酶活性高度依赖于Cu ²⁺与DNA之间的结合化学计量。在H ₂ O ₂介导的3,3'，5,5'-四甲基联苯胺的氧化中，在AG ₃（T ₂ AG ₃）₃存在下，Cu ²⁺的催化活性可以提高约88倍，而在单核苷酸（GMP）的存在下，它显示出轻微的增强。Cu ²⁺的配位具有DNA结构的分子大大增强了其产生活性氧的能力。这项研究中最有趣的发现是，DNA-Cu（II）复合物在被S1核酸酶切割DNA支架后失去了酶活性。建立了比色法，仅使用无标记的寡核苷酸和铜离子即可进行选择性评估S1核酸酶活性，检出限为4.7×10 ^-3 U mL ^-1，而无需复杂的合成和装置。这种简便的方法也可适用于生物流体中的定量分析。

更新日期：2018-02-13

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