在ATP结合的DNA Aptamer中的G-四重体结构强烈调节了连接体结合活性
Aleah N Edwards1, Alexandria N Iannucci1, Jacob VanDenBerg1
1Northern Michigan University, 1401 Presque Isle Ave, Marquette, Michigan 49855, United States.
ACS omega
|April 1, 2024
概括
DNA 体形成G-四重复结构,用于结合腺三酸盐 (ATP) 和 thioflavin T (ThT). 修改后的体表现出增强的ATP结合和双联体识别,使新的分子工具成为可能.
科学领域:
- 分子生物学分子生物学
- 生物化学 生物化学
- 生物技术是生物技术.
背景情况:
- DNA 体形成了用于小分子联体结合的二次结构.
- 这些结构是稳定的,对于开发诊断和监测中的分子工具是有用的.
- 亚丁三酸盐 (ATP) 亚体因其结合能力而受到探索.
研究的目的:
- 为了研究腺三酸盐 (ATP) 结合的阿胺,以同时检测ATP和硫黄素T (ThT).
- 探索G-四重复 (G4) 基因在调节阿巴胺体连接体结合性质中的作用.
- 为了设计具有增强结合亲和力和双联体识别的aptamer变体.
主要方法:
- 使用光标签和无标签的检测方法.
- 构建了带有增加G-四重复单元 (三或四个G-四重奏) 的扩展的阿普坦变体.
- 进行了平衡结合试验和电喷离子质谱学 (ESI-MS) 分析.
主要成果:
- 原生体形成一个G-四重复 (G4) 结构,有两个G-四重复,两者都参与结合.
- 扩展的G4体变体与原生体 (0.15μM) 相比,对ATP表现出明显更强的亲和力 (Kd范围为0.040-0.042μM).
- 无标签和ESI-MS分析证实了THT和ATP对工程G4结构的双重结合.
结论:
- 在这些体中,G-四重复 (G4) 基因对于最佳的ATP结合至关重要.
- 调节G4结构增强了ATP结合亲和力,并使多个连接体同时结合.
- 这些发现有助于设计先进的分子工具,用于生物标志物监测和连接体结合研究.
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