Johnson Research Foundation and Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104-6509, United States.
Johnson Research Foundation and Department of Biochemistry and Biophysics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104-6509, United States.
Methods. 2018 Sep 15;148:67-80. doi: 10.1016/j.ymeth.2018.06.012. Epub 2018 Jun 30.
Pressure and temperature are the two fundamental variables of thermodynamics. Temperature and chemical perturbation are central experimental tools for the exploration of macromolecular structure and dynamics. Though it has long been recognized that hydrostatic pressure offers a complementary and often unique view of macromolecular structure, stability and dynamics, it has not been employed nearly as much. For solution NMR applications the limited use of high-pressure is undoubtedly traced to difficulties of employing pressure in the context of modern multinuclear and multidimensional NMR. Limitations in pressure tolerant NMR sample cells have been overcome and enable detailed studies of macromolecular energy landscapes, hydration, dynamics and function. Here we review the practical considerations for studies of biological macromolecules at elevated pressure, with a particular emphasis on applications in protein biophysics and structural biology.
压力和温度是热力学的两个基本变量。温度和化学扰动是探索大分子结构和动力学的核心实验工具。尽管人们早就认识到静水压力提供了对大分子结构、稳定性和动力学的互补且通常是独特的视角，但它的应用并不广泛。对于溶液 NMR 应用，高压的有限应用无疑可以追溯到在现代多核和多维 NMR 背景下使用压力的困难。耐压 NMR 样品池的限制已经克服，使人们能够详细研究大分子的能量景观、水合作用、动力学和功能。在这里，我们综述了在高压下研究生物大分子的实际考虑因素，特别强调了在蛋白质生物物理学和结构生物学中的应用。
