用于研究微秒至毫秒时间尺度上蛋白质构象动力学的15N CPMG弛豫色散
15N CPMG Relaxation Dispersion for the Investigation of Protein Conformational Dynamics on the µs-ms Timescale.
作者信息
Singh Aayushi, Purslow Jeffrey A, Venditti Vincenzo
机构信息
Department of Chemistry, Iowa State University.
Department of Chemistry, Iowa State University; Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University;
出版信息
J Vis Exp. 2021 Apr 19(170). doi: 10.3791/62395.
Abstract
Protein conformational dynamics play fundamental roles in regulation of enzymatic catalysis, ligand binding, allostery, and signaling, which are important biological processes. Understanding how the balance between structure and dynamics governs biological function is a new frontier in modern structural biology and has ignited several technical and methodological developments. Among these, CPMG relaxation dispersion solution NMR methods provide unique, atomic-resolution information on the structure, kinetics, and thermodynamics of protein conformational equilibria occurring on the µs-ms timescale. Here, the study presents detailed protocols for acquisition and analysis of a N relaxation dispersion experiment. As an example, the pipeline for the analysis of the µs-ms dynamics in the C-terminal domain of bacteria Enzyme I is shown.
摘要
蛋白质构象动力学在酶催化、配体结合、别构作用和信号传导等调节过程中发挥着重要作用，这些都是重要的生物学过程。了解结构与动力学之间的平衡如何支配生物学功能是现代结构生物学的一个新前沿领域，并引发了多项技术和方法的发展。其中，CPMG弛豫色散溶液核磁共振方法提供了关于在微秒至毫秒时间尺度上发生的蛋白质构象平衡的结构、动力学和热力学的独特原子分辨率信息。在此，本研究展示了用于获取和分析¹⁵N弛豫色散实验的详细方案。作为一个例子，展示了细菌酶I C端结构域中微秒至毫秒动力学分析的流程。
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