Roy J. Carver Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USA.
Department of Biological Sciences, Rensselaer Polytechnic Institute, Troy, NY 12180, USA.
Prog Nucl Magn Reson Spectrosc. 2017 Nov;102-103:15-31. doi: 10.1016/j.pnmrs.2017.05.003. Epub 2017 Jun 2.
High-pressure is a well-known perturbation method used to destabilize globular proteins. It is perfectly reversible, which is essential for a proper thermodynamic characterization of a protein equilibrium. In contrast to other perturbation methods such as heat or chemical denaturant that destabilize protein structures uniformly, pressure exerts local effects on regions or domains of a protein containing internal cavities. When combined with NMR spectroscopy, hydrostatic pressure offers the possibility to monitor at a residue level the structural transitions occurring upon unfolding and to determine the kinetic properties of the process. High-pressure NMR experiments can now be routinely performed, owing to the recent development of commercially available high-pressure sample cells. This review summarizes recent advances and some future directions of high-pressure NMR techniques for the characterization at atomic resolution of the energy landscape of protein folding.
高压是一种众所周知的用于使球状蛋白质不稳定的扰动方法。它是完全可逆的，这对于蛋白质平衡的适当热力学特性描述是必不可少的。与其他扰动方法（如均匀地使蛋白质结构不稳定的热或化学变性剂）不同，压力对含有内部空腔的蛋白质区域或结构域施加局部效应。当与 NMR 光谱学结合使用时，静水压力提供了在残基水平上监测变性过程中发生的结构转变并确定该过程的动力学性质的可能性。由于最近开发了商业上可用的高压样品池，现在可以常规地进行高压 NMR 实验。这篇综述总结了高压 NMR 技术在原子分辨率下对蛋白质折叠的能量景观进行描述的最新进展和一些未来方向。
