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基于毛细管筛分电泳评估小分子与 DNA 结合导致的结构变化。

Assessment of the structural change of DNA by binding with a small molecule based on capillary sieving electrophoresis.

机构信息

Department of Applied Chemistry, Graduate School of Engineering, Osaka Metropolitan University, Osaka, Japan.

CREST, Japan Science and Technology Agency, Tokyo, Japan.

出版信息

Anal Sci. 2024 Apr;40(4):773-780. doi: 10.1007/s44211-024-00524-8. Epub 2024 Feb 28.

Abstract

In this study, capillary sieving electrophoresis (CSE) using polymer solutions was used to evaluate the structural changes in nucleic acids upon complexation with small molecules. As the model target and nucleic acids, L-tyrosinamide (Tyr-Am) and its aptamer, which is a type of DNA specifically binding to Tyr-Am, were selected. CSE was conducted using a capillary filled with background solution (BGS) containing hydroxypropyl cellulose (HPC) as a sieving matrix. When Tyr-Am or tyrosine was added to the BGS in CSE, the ratio of mobility differences of the Tyr-Am-aptamer complex increased compared to that of the free aptamer without the addition of Tyr-Am. In contrast, when other amino acids or their analogs were added, results showed no apparent change or decreases in electrophoretic mobility. These results indicate that the proposed method can be applied to assess structural changes in nucleic acids that target small molecules.

摘要

在这项研究中,使用聚合物溶液的胶束筛分电泳(CSE)来评估小分子与核酸复合时核酸结构的变化。选择 L-酪氨酸酰胺(Tyr-Am)及其适体作为模型靶标和核酸,适体是一种专门与 Tyr-Am 结合的 DNA 类型。CSE 使用充满包含羟丙基纤维素(HPC)作为筛分基质的背景溶液(BGS)的毛细管进行。当 Tyr-Am 或酪氨酸被添加到 CSE 中的 BGS 中时,与未添加 Tyr-Am 的游离适体相比,Tyr-Am-适体复合物的迁移率差异比增加。相比之下,当添加其他氨基酸或其类似物时,结果显示电泳迁移率没有明显变化或降低。这些结果表明,所提出的方法可用于评估针对小分子的核酸结构变化。

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