Crystallography and Molecular Biology Division, Saha Institute of Nuclear Physics, A CI of Homi Bhabha National Institute, 1/AF Bidhannagar, Kolkata 700 064, India.
STAR Protoc. 2022 Feb 28;3(1):101200. doi: 10.1016/j.xpro.2022.101200. eCollection 2022 Mar 18.
Comprehensive understanding of a protein's function depends on having reliable, sophisticated tools to study protein structural dynamics in physiologically-relevant conditions. Here, we present an effective, robust step-by-step protocol to monitor the structural dynamics (including hydration dynamics) of a protein utilizing various site-directed fluorescence (SDFL) approaches. This protocol should be widely applicable for studying soluble proteins, intrinsically-disordered proteins, and membrane proteins. For complete details on the use and execution of this protocol, please refer to Das et al. (2020), Das and Raghuraman (2021), and Chatterjee et al. (2021).
全面了解蛋白质的功能取决于是否有可靠、复杂的工具来研究生理相关条件下的蛋白质结构动力学。在这里，我们提出了一种有效、稳健的逐步方案，利用各种定点荧光（SDFL）方法来监测蛋白质的结构动力学（包括水动力学）。该方案应广泛适用于研究可溶性蛋白、固有无序蛋白和膜蛋白。有关此方案的使用和执行的完整详细信息，请参阅 Das 等人（2020 年）、Das 和 Raghuraman（2021 年）和 Chatterjee 等人（2021 年）。
