Applications of Differential Scanning Fluorometry and Related Technologies in Characterization of Protein-Ligand Interactions
Applications of Differential Scanning Fluorometry and Related Technologies in Characterization of Protein-Ligand Interactions
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DOI：
10.1007/978-1-0716-0163-1_4
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发表时间：
2020-01-01
期刊：
影响因子：
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通讯作者：
Simeonov, Anton
中科院分区：
文献类型：
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作者：
Baljinnyam, Bolormaa;Ronzetti, Michael;Simeonov, Anton
差示扫描荧光测定 (DSF) 是一种高效、高通量的方法，可分析蛋白质稳定性，并通过扰动蛋白质熔解温度来检测配体相互作用。该方法通过观察样品的荧光变化来监测蛋白质的去折叠，无论是通过环境敏感的荧光团还是通过内在的蛋白质荧光，同时施加温度梯度。在这里，我们详细描述了如何开发和优化 DSF 测定，以识别蛋白质-配体相互作用，同时探索不同的缓冲液和添加剂条件。还讨论了数据分析和该方法的进一步应用。
Differential scanning fluorometry (DSF) is an efficient and high-throughput method to analyze protein stability, as well as detect ligand interactions through perturbations of the protein's melting temperature. The method monitors protein unfolding by observing the fluorescence changes of a sample, whether through an environmentally sensitive fluorophore or by intrinsic protein fluorescence, while a temperature gradient is applied. Here, we describe in detail how to develop and optimize DSF assays to identify protein-ligand interactions while exploring different buffer and additive conditions. Analysis of the data and further applications of the method are also discussed.