Department of Chemistry, York University, Toronto, ON, Canada.
Département de chimie, Université de Sherbrooke, Sherbrooke, QC, Canada.
Q Rev Biophys. 2024 Sep 3;57:e9. doi: 10.1017/S0033583524000076.
Structure-switching aptamers have become ubiquitous in several applications, notably in analytical devices such as biosensors, due to their ease of supporting strong signaling. Aside from their ability to bind specifically with their respective target, this class of aptamers also undergoes a conformational rearrangement upon target recognition. While several well-studied and early-developed aptamers (e.g., cocaine, ATP, and thrombin) have been found to have this structure-switching property, the vast majority do not. As a result, it is common to try to engineer aptamers into switches. This proves challenging in part because of the difficulty in obtaining structural and functional information about aptamers. In response, we review various readily available biophysical characterization tools that are capable of assessing structure switching of aptamers. In doing so, we delve into the fundamentals of these different techniques and detail how they have been utilized in characterizing structure-switching aptamers. While each of these biophysical techniques alone has utility, their real power to demonstrate the occurrence of structural change with ligand binding is when multiple techniques are used. We hope that through a deeper understanding of these techniques, researchers will be better able to acquire biophysical information about their aptamer-ligand systems and accelerate the translation of aptamers into biosensors.
结构转换适体在多个应用领域中已经变得无处不在，特别是在分析设备（如生物传感器）中，因为它们易于支持强信号。除了能够特异性地与各自的靶标结合之外，这一类适体在靶标识别时还会发生构象重排。虽然已经发现了一些经过充分研究和早期开发的适体（例如可卡因、ATP 和凝血酶）具有这种结构转换特性，但绝大多数适体并不具有这种特性。因此，人们通常试图将适体设计成开关。这在一定程度上具有挑战性，因为获得适体的结构和功能信息具有难度。有鉴于此，我们回顾了各种现有的生物物理特性分析工具，这些工具能够评估适体的结构转换。在进行综述的过程中，我们深入探讨了这些不同技术的基本原理，并详细说明了它们在表征结构转换适体方面的应用。虽然这些生物物理技术本身都具有一定的实用性，但只有当多种技术结合使用时，它们才具有展示配体结合时结构变化的真正力量。我们希望通过对这些技术的更深入理解，研究人员能够更好地获取适体-配体系统的生物物理信息，并加速适体在生物传感器中的转化。
