Astbury Centre for Structural Molecular Biology and School of Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom; email:
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA; email:
Annu Rev Biophys. 2022 May 9;51:223-246. doi: 10.1146/annurev-biophys-090921-120150. Epub 2022 Jan 19.
Molecular chaperones are the guardians of the proteome inside the cell. Chaperones recognize and bind unfolded or misfolded substrates, thereby preventing further aggregation; promoting correct protein folding; and, in some instances, even disaggregating already formed aggregates. Chaperones perform their function by means of an array of weak protein-protein interactions that take place over a wide range of timescales and are therefore invisible to structural techniques dependent upon the availability of highly homogeneous samples. Nuclear magnetic resonance (NMR) spectroscopy, however, is ideally suited to study dynamic, rapidly interconverting conformational states and protein-protein interactions in solution, even if these involve a high-molecular-weight component. In this review, we give a brief overview of the principles used by chaperones to bind their client proteins and describe NMR methods that have emerged as valuable tools to probe chaperone-substrate and chaperone-chaperone interactions. We then focus on a few systems for which the application of these methods has greatly increased our understanding of the mechanisms underlying chaperone functions.
分子伴侣是细胞内蛋白质组的守护者。伴侣蛋白识别并结合未折叠或错误折叠的底物，从而防止进一步聚集；促进正确的蛋白质折叠；在某些情况下，甚至可以解聚已经形成的聚集体。伴侣蛋白通过一系列弱的蛋白质-蛋白质相互作用来发挥其功能，这些相互作用发生在广泛的时间尺度上，因此对于依赖于高度均相样品可用性的结构技术来说是不可见的。然而，核磁共振（NMR）光谱特别适合于研究溶液中动态的、快速相互转化的构象状态和蛋白质-蛋白质相互作用，即使这些相互作用涉及高分子量的成分。在这篇综述中，我们简要概述了伴侣蛋白结合其客户蛋白所使用的原理，并描述了已成为探测伴侣蛋白-底物和伴侣蛋白-伴侣蛋白相互作用的有价值工具的 NMR 方法。然后，我们将重点介绍一些应用这些方法极大地提高了我们对伴侣蛋白功能机制的理解的系统。
