Jones Christopher P, Piszczek Grzegorz, Ferré-D'Amaré Adrian R
Biochemistry and Biophysics Center, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
Biophysics Core Facility, National Heart, Lung and Blood Institute, National Institutes of Health, Bethesda, MD, USA.
Methods Mol Biol. 2019;1964:75-87. doi: 10.1007/978-1-4939-9179-2_6.
One of the many ways by which bacteria control gene expression is through cis-acting regulatory mRNA elements called riboswitches. By specifically binding to small molecules or metabolites and pairing the binding event to an RNA structure change, riboswitches link a metabolic input to a transcriptional or translational output. For over a decade, isothermal titration calorimetry (ITC) has been used to investigate how riboswitches interact with small molecules. We present methods for assaying RNA-ligand interactions using ITC and analyzing resulting data to estimate thermodynamic parameters associated with binding.
细菌控制基因表达的众多方式之一是通过称为核糖开关的顺式作用调节mRNA元件。核糖开关通过特异性结合小分子或代谢物，并将结合事件与RNA结构变化配对，将代谢输入与转录或翻译输出联系起来。十多年来，等温滴定量热法（ITC）一直用于研究核糖开关如何与小分子相互作用。我们介绍了使用ITC测定RNA-配体相互作用并分析所得数据以估计与结合相关的热力学参数的方法。
