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100 kHz magic angle spinning for development of solid-state NMR methodology for probing protein dynamics

100 kHz 魔角旋转用于开发探测蛋白质动力学的固态 NMR 方法

基本信息

批准号：
EP/L025906/1
负责人：
Józef Lewandowski
金额：
$ 14.4万
依托单位：
University of Warwick
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2014
资助国家：
英国
起止时间：
2014 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=EP%2FL025906%2F1
关键词：
100 kHz magic angle spinning

项目摘要

Motion and change are essential features of living organisms and fundamentally important for many vital processes from protein folding and unfolding, ligand binding, signalling, allosteric regulation to enzymatic catalysis. Consequently, understanding motions at molecular level provides valuable insights into the phenomena involving change of structure both when they function as intended or when they malfunction. For example understanding how proteins misfold may help to fight debilitating diseases called amyloidoses that include Alzheimer's disease, type II diabetes or bovine spongiform encephalopathy more widely known as "mad cow" disease. Moreover, understanding motions that are intrinsically associated with signalling pathways may result in development of better drugs that target such pathways (most medicines work this way). Even development of practical environmentally friendly biobatteries and biofuel cells may be aided by knowledge of molecular motions as they make use of enzymes. Thus it is really important to devise ways to measure protein motions at atomic resolution.To do that, in this project, we will develop a technique called nuclear magnetic resonance (NMR), which relies on the inherent magnetism of atomic nuclei. When placed in a strong magnetic field magnetic moments of nuclei align with the external field but this alignment may be changed by application of radio waves at specific frequencies. By measuring the associated frequencies one can learn about the relative position of atoms with respect to each other and how this position changes with time i.e. molecular motions. A very powerful aspect of this technique is that one can learn such information not only for a molecule overall but for specific atoms in it. In solid-state NMR, which is the primary method used in this project, the high resolution necessary to distinguish individual sites is enabled by a technique called magic angle spinning (MAS), which involves fast rotation of the sample around an axis inclined at an angle of 54.7 degrees to the external magnetic field. Recently introduced cutting edge instrumentation allows achieving spinning frequencies up to 100 000 revolutions per second. The centre of this project is the purchase of the first in the UK probe capable of 100 kHz MAS. The improved efficiency of MAS at such astounding frequencies makes possible designing new experiments that provide new analytical tools to access motions, e.g. site-specific 1H relaxation or highly sensitive 1H-detected relaxation measurements in fully protonated samples. The 100 kHz spinning removes a number of undesired effects obscuring the measurements of parameters reporting on molecular motions and thus allows a detailed view of protein motions to be obtained.In this project we propose to develop a series of robust solid-state NMR spectroscopic methods that take advantage of the new 100 kHz spinning regime and will provide improved access to measuring of dynamic processes in proteins at atomic resolution and in a site-specific manner. In particular, we will focus on techniques that provide access to slow motions in the regime that is difficult to access by the solid-state NMR sister method - solution NMR. In addition, in order to improve practicality of the developed techniques we will optimise them for speed and sensitivity.

运动和变化是生物体的基本特征，对于从蛋白质折叠和解折叠、配体结合、信号传导、变构调节到酶催化的许多重要过程至关重要。因此，在分子水平上理解运动可以为涉及结构变化的现象提供有价值的见解，无论是当它们按预期发挥作用还是当它们发生故障时。例如，了解蛋白质如何错误折叠可能有助于对抗称为淀粉样变性的衰弱性疾病，包括阿尔茨海默病、II 型糖尿病或牛海绵状脑病（更广泛地称为“疯牛”病）。此外，了解与信号传导途径本质上相关的运动可能会导致开发出针对这些途径的更好的药物（大多数药物都是这样工作的）。即使是实用的环保生物电池和生物燃料电池的开发也可以通过利用酶时的分子运动知识来帮助。因此，设计以原子分辨率测量蛋白质运动的方法非常重要。为此，在本项目中，我们将开发一种称为核磁共振 (NMR) 的技术，该技术依赖于原子核的固有磁性。当置于强磁场中时，原子核的磁矩与外部场对齐，但这种对齐可以通过应用特定频率的无线电波来改变。通过测量相关频率，人们可以了解原子之间的相对位置以及该位置如何随时间（即分子运动）变化。这项技术的一个非常强大的方面是，人们不仅可以了解整个分子的信息，还可以了解其中特定原子的信息。在该项目使用的主要方法固态核磁共振中，区分各个位点所需的高分辨率是通过一种称为魔角旋转 (MAS) 的技术实现的，该技术涉及样品绕与外部磁场成 54.7 度倾斜的轴快速旋转。最近推出的尖端仪器可以实现高达每秒 100 000 转的旋转频率。该项目的核心是在英国购买第一个能够支持 100 kHz MAS 的探头。 MAS 在如此惊人的频率下效率的提高使得设计新实验成为可能，这些实验提供了新的分析工具来获取运动，例如在完全质子化的样品中进行位点特异性 1H 弛豫或高灵敏度 1H 检测弛豫测量。 100 kHz 旋转消除了许多干扰报告分子运动参数测量的不良影响，从而获得蛋白质运动的详细视图。在这个项目中，我们建议开发一系列强大的固态核磁共振波谱方法，利用新的 100 kHz 旋转机制，并将提供改进的方法来测量原子分辨率和蛋白质动态过程。特定于站点的方式。特别是，我们将重点关注能够获得固态 NMR 姐妹方法（溶液 NMR）难以获得的慢运动的技术。此外，为了提高所开发技术的实用性，我们将对它们的速度和灵敏度进行优化。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Unraveling the complexity of protein backbone dynamics with combined (13)C and (15)N solid-state NMR relaxation measurements

通过组合 (13)C 和 (15)N 固态 NMR 弛豫测量揭示蛋白质主链动力学的复杂性

DOI：
10.5451/unibas-ep39973
发表时间：
2015
期刊：
影响因子：
0
作者：
Lamley, Jonathan M.
通讯作者：
Lamley, Jonathan M.

1H line width dependence on MAS speed in solid state NMR - Comparison of experiment and simulation.

DOI：
10.1016/j.jmr.2018.04.003
发表时间：
2018-06
期刊：
Journal of magnetic resonance
影响因子：
2.2
作者：
U. Sternberg;R. Witter;I. Kuprov;Jonathan M. Lamley;Andres Oss;Józef R. Lewandowski;A. Samoson
通讯作者：
U. Sternberg;R. Witter;I. Kuprov;Jonathan M. Lamley;Andres Oss;Józef R. Lewandowski;A. Samoson

Dipolar Order Parameters in Large Systems With Fast Spinning.