Funke Andrea, Weisz Klaus
Institute of Biochemistry, University of Greifswald, Greifswald, Germany.
Methods Mol Biol. 2019;2035:45-61. doi: 10.1007/978-1-4939-9666-7_3.
The thermodynamic characterization of G4-ligand interactions has shown to be a powerful adjunct to structural information in the rational design and optimization of potent G-quadruplex ligands for use in therapeutics, diagnostics, or other technological applications. Isothermal titration calorimetry (ITC) can resolve energetic contributions to complex formation and constitutes the only available experimental method to directly measure binding enthalpies. A general protocol for using ITC in studies on quadruplex-ligand interactions with details on the experimental setup, data analysis, and potential pitfalls is presented. The methodologies used are illustrated on results obtained from the targeting of a parallel DNA G-quadruplex with a G4-binding indoloquinoline derivative.
G4配体相互作用的热力学表征已被证明是合理设计和优化用于治疗、诊断或其他技术应用的强效G-四链体配体时，结构信息的有力辅助手段。等温滴定量热法（ITC）可以解析对复合物形成的能量贡献，并且是直接测量结合焓的唯一可用实验方法。本文介绍了在四链体-配体相互作用研究中使用ITC的通用方案，包括实验设置、数据分析和潜在陷阱的详细信息。通过用G4结合吲哚喹啉衍生物靶向平行DNA G-四链体所获得的结果，说明了所使用的方法。
