Systematic Approach to DNA Aptamer Design Using Amino Acid?Nucleic Acid Hybrids (ANHs) Targeting Thrombin
Systematic Approach to DNA Aptamer Design Using Amino Acid?Nucleic Acid Hybrids (ANHs) Targeting Thrombin
复制标题
使用靶向凝血酶的氨基酸核酸杂交体 (ANH) 设计 DNA 适体的系统方法
DOI：
10.1021/acsbiomaterials.1c00060
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发表时间：
2021
影响因子：
5.8
通讯作者：
Park Soyoung
中科院分区：
文献类型：
--
作者：
Yum Ji Hye;Ishizuka Takumi;Fukumoto Koyuki;Hori Daisuke;Bao Hong-Liang;Xu Yan;Sugiyama Hiroshi;Park Soyoung
先天DNA/RNA适配子的化学修饰有助于其功能的改善。在这里，我们报告了我们的模块化策略来操纵凝血酶结合DNA适配子(TBA)来提高其抗凝血活性和结合亲和力。合成了一系列氨基酸结合物，称为氨基酸-核酸杂交物或ANHs，并将其并入TBA环序列。我们发现，在环区用疏水氨基酸取代具有显著增强的抗凝血酶活性，比天然TBA高3倍。我们利用圆二色谱、表面等离子体共振分析和分子模拟等实验技术研究了凝血酶结合亲和力与我们的氨基酸结合物的特性之间的相关性。本研究展示了一种基于氨基酸特性的系统的适配子设计方法，从而允许开发先进的适配子。
Chemical modifications of innate DNA/RNA aptamers facilitate the improvement of their function. Herein, we report our modular strategy to manipulate a thrombin-binding DNA aptamer (TBA) to improve its anticoagulation activity and binding affinity. A set of amino acid conjugates, termed amino acid–nucleic acid hybrids or ANHs, was synthesized and incorporated into a TBA loop sequences. We found that substitutions with hydrophobic amino acids in the loop region possessed significantly enhanced antithrombin activity, up to 3-fold higher than the native TBA. We investigated the correlations between thrombin-binding affinity and the features of our amino-acid conjugates using experimental techniques including circular dichroism spectroscopy, surface plasmon resonance assay, and molecular modeling. The present study demonstrates a systematic approach to aptamer design based on amino-acid characteristics, allowing the development of advanced aptamers.