Amano Ryo, Furukawa Tomohisa, Sakamoto Taiichi
Faculty of Advanced Engineering, Department of Life Science, Chiba Institute of Technology, Narashino-shi, Chiba, Japan.
Methods Mol Biol. 2019;1964:119-128. doi: 10.1007/978-1-4939-9179-2_9.
Aptamers are nucleic acid molecules that bind to a target molecule with high affinity and specificity, which are generated by a process known as systematic evolution of ligands by exponential enrichment (SELEX). Because of their high affinity and specificity, aptamers were developed as therapeutic agents. Although aptamers are investigated as promising therapeutic agents, the mechanism of their high affinity and specificity is not clear. Therefore, structural and biophysical studies are important to know that. To date, ITC is increasingly being used to study the thermodynamic basis of aptamer-target protein interactions. Understanding the mechanism of aptamer binding would contribute to their development for therapeutic applications. In this chapter, we describe the protocol to study the thermodynamics of aptamer-protein interactions.
适体是一类核酸分子，它们能以高亲和力和特异性结合靶分子，是通过一种称为指数富集配体系统进化（SELEX）的过程产生的。由于其高亲和力和特异性，适体被开发为治疗剂。尽管适体作为有前景的治疗剂正在被研究，但其高亲和力和特异性的机制尚不清楚。因此，结构和生物物理研究对于了解这一点很重要。迄今为止，等温滴定量热法（ITC）越来越多地用于研究适体与靶蛋白相互作用的热力学基础。了解适体结合的机制将有助于其在治疗应用中的开发。在本章中，我们描述了研究适体与蛋白质相互作用热力学的实验方案。
