单分子磁镊揭示了 TAV2b 衍生肽对双链 RNA 的下卷和稳定作用

Zainab M. Rashid, Salina Quack, Misha Klein, Quinte Smitskamp, Pim America, Marvin Albers, Jannik Paulus, Tom Grossmann, David Dulin
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引用次数: 0

摘要

双链 RNA(dsRNA)已发展成为了解和调节生物过程的重要工具,在治疗方面具有广阔的前景。然而,由于其在生物环境中的不稳定性,其疗效往往受到限制。最近,从天然 RNA 结合蛋白中提取的多肽 dsRNA 结合剂的开发成为解决这一限制的有利起点。然而,这些高亲和力 dsRNA 粘合剂如何改变 dsRNA 的结构和灵活性仍不清楚。为此,我们采用单分子磁镊实验来研究 TAV2b 衍生的多肽 dsRNA 粘合剂对 dsRNA 机械特性的影响。扭力谱测定表明,这些多肽在稳定双链的同时,还能使dsRNA下卷。此外,力谱实验证明,野生型 TAV2b 肽衍生物能延长轮廓长度并降低 dsRNA 的弯曲刚度,而同源二聚体版本则能在力低于 1 pN 时引发高阶复合物的形成。我们的研究提出了一种定量方法来研究这些肽如何改变 dsRNA 的结构,以及肽的结构设计是否会改变与 dsRNA 的亲和力及其稳定性。这种方法可以为设计更强效、更有效的dsRNA结合剂提供信息,从而推动RNA疗法的发展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Single-molecule magnetic tweezers reveals that TAV2b-derived peptides underwind and stabilize double-stranded RNA
Double-stranded RNA (dsRNA) has evolved into a key tool in understanding and regulating biological processes, with promising implications in therapeutics. However, its efficacy is often limited due to instability in biological settings. Recently, the development of peptidic dsRNA binders derived from naturally occurring RNA-binding proteins has emerged as a favorable starting point to address this limitation. Nevertheless, it remains unclear how these high affinity dsRNA binders alter the structure and flexibility of dsRNA. To this end, we employed single-molecule magnetic tweezers experiments to investigate the effects of TAV2b-derived peptidic dsRNA binders on the mechanical properties of dsRNA. Torque spectroscopy assays demonstrated that these peptides underwind dsRNA, while also stabilizing the duplex. Additionally, force spectroscopy experiments demonstrate that a wild type TAV2b peptide derivative extends the contour length and lowers the bending rigidity of dsRNA, while a homodimeric version triggers the formation of higher order complexes at forces below 1 pN. Our study presents a quantitative approach to investigate how these peptides alter the structure of dsRNA, and whether peptide structural design alters the affinity to dsRNA and its stability. This approach could inform the design of more potent and effective dsRNA binders in the efforts to advance RNA therapeutics.
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