Probing drug receptor binding sites driven by solid state NMR - An interdisciplinary approach.

生物学在原子尺度上通过高度同步的化学作用发挥作用。生物分子电荷的微小变化，甚至氢原子的位置，都可能产生深远的后果(例如，两种微妙不同的沙利度胺(α，β)的作用截然不同，或者血红蛋白中的一种氨基酸发生变化，导致镰状细胞性贫血)。这种微妙之处在分子水平上得到了更好的理解，但在促进健康和福祉方面仍有许多需要发现和理解。膜蛋白是未来十年疾病控制的主要靶点。这些是细胞与外界的第一个接触点，大约85%的细胞信号是通过膜传递的。因此，学者和制药公司都对这种信号如何传递到细胞内感兴趣也就不足为奇了(2005年，两个诺贝尔奖获奖，原因是发现了一个靶标膜蛋白可以激活并向许多其他蛋白质发出信号，这取决于小分子激活的性质，自1987年以来，膜蛋白研究获得了10多个诺贝尔奖)。膜蛋白很难处理，这就是为什么数据库中只有21个结构(数百万个可用结构)。此外，我们没有任何配体激活的人类受体的结构。我们现在需要的是详细了解这些信号是如何在分子水平上启动和传输的，这可以使用专门为探测细节而设计的核磁共振方法来解决，分辨率非常高(优于0.03纳米)，具有电子和动态细节，但非常重要的是，在缺乏目标受体蛋白的总结构的情况下。固态核磁共振特别利用某些特定原子的磁性来处理大的异质、无序大分子-这一直是结构生物学中一个快速增长的领域，英国处于发展的前沿。这项工作的一个重要部分是将磁性间谍(或标记)结合到感兴趣的分子中，以便我们能够获得所需的信息。监测核的化学插入到大分子中信息丰富的位置是至关重要的，也是先决条件，只能来自最先进的巧妙化学，目的是使用物理方法回答生物学上的重要问题。核磁共振方法在在信息丰富的位置产生非常局部化和高度特定的信息方面是独一无二的，但这只有通过使用高度专业化的化学来在需要的地方制造具有核磁共振标记的分子才可能实现-因此，这项资金申请将结合这两个专业领域(牛津的核磁共振和布里斯托尔的标记)来回答重要的生物学问题，即小分子如何激活蛋白质以将信号传输到细胞中？获得的详细信息将有助于理解，例如，一种激素如何引起特定的反应，或一种有毒化学物质如何启动细胞死亡。对于传统上在药物发现方面非常成功的英国来说，新的设计原则将被阐明，这对创造财富非常重要。

Biology works through highly synchronised chemical interactions at the atomistic scale. Small changes in the electronic charge of a biological molecule, or even the position of a hydrogen atom can have far-reaching consequences (e.g. the very contrasting actions of two subtly different forms (alpha, beta) of thalidomide, or a change of one amino acid in haemoglobin causing sickle cell anaemia). Such subtleties are becoming better understood at the molecular level, but still much is to be discovered and understood for promotion of health and well-being. The major class of targets in disease control for the next ten years is membrane proteins. These are a cell's first point of contact with the outside world and about 85% of all signals to the cell are transmitted through the membrane. It is not surprising then that both academics and drug companies are interested in how such signals are transmitted into the cell (2 Nobel prizes were awarded in 2005 for the revelation that one target membrane protein can activate and signal a multitude of other proteins, depending upon the nature of the small molecule activation, and over 10 Nobel prizes have been awarded for membrane protein studies since 1987). Membrane proteins are very difficult to work with, which is why there are only 21 structures (out of millions available) in the data bases. In addition, we do not have the structure of any ligand-activated human receptor. What we now need is a detailed insight into how these signals are initiated and transmitted at the molecular level, and this can be addressed using nuclear magnetic resonance (NMR) methods designed specifically for probing the detail at very high resolution (better than 0.03 nanometres) and with electronic and dynamic details but, very importantly, in the absence of the total structure of the target receptor protein.Solid state NMR exploits specifically the magnetic properties of some specific atoms for large heterogeneous, non-ordered macromolecules - this has been a fast growing area in structural biology and the UK is at the forefront of the developments. An essential part of this work is the incorporation of magnetic spies (or labels) into the molecule of interest so that we can obtain the information required. The chemical insertion of monitoring nuclei into the information-rich position in the macromolecule is vital and a pre-requisite and can only come from state-of-the-art clever chemistry directed at answering biologically important questions using physical methods. The NMR method is unique in producing very localized and highly specific information at a information-rich site, but this is only possible through the use of highly specialised chemistry to make molecules with the NMR labels where needed - hence this funding application will combine these two areas of expertise (NMR at Oxford and labelling at Bristol) to answer the important biological question How do small molecules activate proteins to transmit signals into a cell? . Detailed information gained will facilitate the understanding of, e.g. how a hormone causes a particular response, or how a toxic chemical initiates cell death. Importantly for wealth creation for the UK, which traditionally has been highly successful in discovering drugs, new design principles will be elucidated.