Department of Molecular Genetics, University of Toronto, Toronto, ON, M5S 1A8, Canada.
Department of Biochemistry, University of Toronto, Toronto, ON, M5S 1A8, Canada.
Proc Natl Acad Sci U S A. 2021 Dec 14;118(50). doi: 10.1073/pnas.2116325118.
Developments in solution NMR spectroscopy have significantly impacted the biological questions that can now be addressed by this methodology. By means of illustration, we present here a perspective focusing on studies of a number of molecular machines that are critical for cellular homeostasis. The role of NMR in elucidating the structural dynamics of these important molecules is emphasized, focusing specifically on intersubunit allosteric communication in homo-oligomers. In many biophysical studies of oligomers, allostery is inferred by showing that models specifically including intersubunit communication best fit the data of interest. Ideally, however, experimental studies focusing on one subunit of a multisubunit system would be performed as an important complement to the more traditional bulk measurements in which signals from all components are measured simultaneously. Using an approach whereby asymmetric molecules are prepared in concert with NMR experiments focusing on the structural dynamics of individual protomers, we present examples of how intersubunit allostery can be directly observed in high-molecular-weight protein systems. These examples highlight some of the unique roles of solution NMR spectroscopy in studies of complex biomolecules and emphasize the important synergy between NMR and other atomic resolution biophysical methods.
溶液 NMR 光谱学的发展极大地影响了现在可以通过该方法解决的生物学问题。通过举例，我们在这里提出了一个观点，重点关注对许多对细胞内稳态至关重要的分子机器的研究。强调了 NMR 在阐明这些重要分子结构动力学方面的作用，特别关注同寡聚体中亚基间的变构通讯。在寡聚体的许多生物物理研究中，通过显示专门包括亚基间通讯的模型最能拟合感兴趣的数据来推断变构作用。然而，理想情况下，针对多亚基系统的一个亚基进行的实验研究将作为对更传统的整体测量的重要补充，其中同时测量所有成分的信号。通过采用一种方法，使不对称分子与 NMR 实验同时制备，该实验集中于单个原聚体的结构动力学，我们展示了如何在高分子量蛋白质系统中直接观察亚基间变构作用的例子。这些例子突出了溶液 NMR 光谱学在复杂生物分子研究中的一些独特作用，并强调了 NMR 和其他原子分辨率生物物理方法之间的重要协同作用。
