Architecture et Réactivité de l'ARN - CNRS UPR 9002, Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, Strasbourg, France.
Methods Mol Biol. 2020;2113:237-250. doi: 10.1007/978-1-0716-0278-2_16.
Isothermal titration calorimetry (ITC) provides a sensitive, powerful, and accurate tool to suitably analyze the thermodynamic of RNA binding events. This approach does not require any modification or labeling of the system under analysis and is performed in solution. ITC is a very convenient technique that provides an accurate determination of binding parameters, as well as a complete thermodynamic profile of the molecular interactions. Here we show how this approach can be used to characterize the interactions between the dimerization initiation site (DIS) RNA localized within the HIV-1 viral genome and aminoglycoside antibiotics. Our ITC study showed that the 4,5-disubstituted 2-desoxystreptamine (2-DOS) aminoglycosides can bind the DIS with a nanomolar affinity and a high specificity.
等温滴定量热法（ITC）为分析 RNA 结合事件的热力学性质提供了一种灵敏、强大且精确的工具。该方法不需要对分析系统进行任何修饰或标记，并且在溶液中进行。ITC 是一种非常方便的技术，它可以准确确定结合参数，并提供分子相互作用的完整热力学概况。在这里，我们展示了如何使用这种方法来表征定位于 HIV-1 病毒基因组内的二聚化起始位点（DIS）RNA 与氨基糖苷类抗生素之间的相互作用。我们的 ITC 研究表明，4,5-二取代 2-去氧链霉胺（2-DOS）氨基糖苷类可以以纳摩尔亲和力和高特异性与 DIS 结合。
