Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 34141, South Korea.
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Daejeon 34141, South Korea.
Mol Cells. 2024 Oct;47(10):100112. doi: 10.1016/j.mocell.2024.100112. Epub 2024 Sep 16.
The determination of the dissociation constant (K) is pivotal in biochemistry and pharmacology for understanding binding affinities in chemical reactions, which is crucial for drug development and comprehending biological systems. Here, we introduce a single-molecule fluorescence resonance energy transfer-based method for determining K, alongside the conventional electrophoretic mobility shift assay method of K, offering insights into thermodynamic interactions between proteins and substrates. The single-molecule fluorescence resonance energy transfer approach is highlighted for its ability to accurately measure binding and dissociation kinetics through fluorescence labeling and the intrinsic nature of protein-DNA interactions, representing a significant advancement in the fields of molecular biology and pharmacology.
在生物化学和药理学中，确定离解常数 (K) 对于理解化学反应中的结合亲和力至关重要，这对于药物开发和理解生物系统至关重要。在这里，我们介绍了一种基于单分子荧光共振能量转移的 K 测定方法，以及传统的电泳迁移率变动分析方法的 K 测定方法，为蛋白质和底物之间的热力学相互作用提供了深入的了解。单分子荧光共振能量转移方法通过荧光标记和蛋白质-DNA 相互作用的固有性质，突出了其准确测量结合和解离动力学的能力，是分子生物学和药理学领域的重大进展。
