Recent advances in solid-state nuclear magnetic resonance techniques to quantify biomolecular dynamics.
Recent advances in solid-state nuclear magnetic resonance techniques to quantify biomolecular dynamics.
复制标题
量化生物分子动力学的固态核磁共振技术的最新进展。
DOI：
10.1021/ac403956k
复制
发表时间：
2014
影响因子：
7.4
通讯作者：
Rienstra,ChadM
中科院分区：
文献类型：
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作者：
Watt,EricD;Rienstra,ChadM
过去十年来，固态核磁共振（SSNMR）技术的出现极大地扩大了适合结构研究的可及生物分子的范围。1，2这些方法被开发并首次应用于各种微晶模型蛋白，包括BPTI，3 SH 3，4，5泛素，6，7 kaliotoxin，8和GB 1。这些研究表明，整个蛋白质中的单个13 C、15 N和1H位点可以被唯一地分辨和分配，以实现高分辨率结构测定的目的。传统的固态NMR方法需要位点特异性同位素标记，而新的发展使整个蛋白质能够通过在多个维度上解析信号来进行检查。这些功能还允许通过相关光谱法测量动态参数，这增加了这种测量的吞吐量、灵敏度、可靠性和再现性。SSNMR的一个主要优点是可以检查大范围的样品条件和类型，包括许多溶液NMR方法无法获得的样品，例如膜蛋白和原纤维，以及微晶。最近的例子包括HET-s朊病毒的原纤维，12 α B-晶状体蛋白的高分子量寡聚体，13 M2肽与结合的金刚烷胺药物在生理学相关双层中的四聚体组装，14三聚体膜蛋白YadA，15光感受器蛋白视紫红质，16甲状腺素运载蛋白的核心结构域的原纤维，17和源自阿尔茨海默病脑的β-淀粉样蛋白斑，疾病患者。18这些研究为探索基础生物物理学和生物化学开辟了新的途径，并使人们深入了解了溶液核磁共振或X射线晶体学无法获得的膜和聚集态中的临床相关事件。此外，SSNMR实验适用于比溶液NMR 19所能达到的温度范围更大的温度范围，从而能够实现从生理温度到大多数晶体结构得以解析的低温温度的连续实验条件。一般来说，NMR方法的另一个优点是它们是非扰动的，并且不需要蛋白质的共价修饰。因此，从这样的实验中获得的动态信息有可能准确地代表天然蛋白质的行为。本文综述了近五年来固体核磁共振动力学研究的方法和应用，研究时间范围从纳秒到几天。
The emergence of solid-state nuclear magnetic resonance (SSNMR) techniques over the past decade has greatly expanded the range of accessible biomolecules amenable to structural study. 1, 2 These methods were developed and first applied to a variety of microcrystalline model proteins, including BPTI, 3 SH3, 4, 5 ubiquitin, 6, 7 kaliotoxin, 8 and GB1. 9− 11 These studies demonstrated that individual 13C, 15N, and 1H sites throughout the entire protein could be uniquely resolved and assigned for purposes of high-resolution structure determination. Whereas traditional solid-state NMR approaches required site-specific isotopic labeling, the new developments enable entire proteins to be examined by resolving signals in multiple dimensions. These capabilities also permit the measurement of dynamic parameters through correlation spectroscopy, which increases the throughput, sensitivity, reliability, and reproducibility of such measurements. A major advantage of SSNMR is that a large range of sample conditions and types can be examined, including many that are not accessible to solution NMR methods, such as membrane proteins and fibrils, in addition to microcrystals. Recent examples include fibrils of the HET-s prion, 12 high molecular weight oligomers of αB-crystallin, 13 the tetrameric assembly of the M2 peptide with the bound amantadine drug in physiologically relevant bilayers, 14 the trimeric membrane protein YadA, 15 the photoreceptor proteorhodopsin, 16 fibrils of the core domain of transthyretin, 17 and beta-amyloid plaques derived from the brains of Alzheimer, s disease patients. 18 Such studies have opened up new avenues for exploration of fundamental biophysics and biochemistry and yielded insights into clinically relevant events in membranes and aggregated states not accessible to solution NMR or X-ray crystallography. Moreover, SSNMR experiments are applicable over a larger range of temperatures than can be accessed by solution NMR 19 and thereby enable a continuum of experimental conditions from physiological temperatures to the cryogenic temperatures at which most crystal structures are solved. Another advantage of NMR methods in general is that they are nonperturbing and do not require covalent modification of the protein. Thus, the dynamic information procured from such experiments has the potential to accurately represent the native protein behavior. This review focuses on developments over the last five years, regarding methods and applications of dynamics studies by solid-state NMR over timescales ranging from nanoseconds to days.