An investigation into the conformational changes and lipid dependence of NTS1 activation by its agonist

我们的许多神经功能是通过驻留在大脑中的受体蛋白（称为gpcr）来控制的，因此，据估计，我们使用的所有药物中约有30%作用于这些受体，其中有800多种。GPCRs控制的功能是多种多样的，一个受体可能参与多种反应。我们需要找出这些受体是如何工作的，据说这是当前结构生物学的主要挑战（Lagerstrom & Schioth, 2008, Nature Reviews Drug Discovery, 7,339 -57）。为了帮助理解这个过程，作为最近的突破的结果，现在有可能制造非常有限数量的gpcr，这样我们就可以开始发现它们是如何起作用的。我们选择的研究对象涉及疼痛管理、神经肌肉疾病（如帕金森氏症）、食欲控制以及肥胖控制。我们已经能够在简单的大肠杆菌细胞中表达这种哺乳动物大脑受体，它在功能上很活跃，我们所做的任何工作都与它在大脑中的功能有关。和许多受体一样，它们是由小分子结合激活的，尽管我们可以监控这种结合，但重要的是要了解和研究蛋白质是如何被激活的，以及它是如何将信号传递给其他蛋白质，最终传递给细胞的。因此，我们将表达和纯化受体，用激素激活它，然后使用最先进的方法来探测受体在激活过程中的变化。这些方法可以用来测量这些纳米探测器在纳米尺度（0.5-8nm +/- 0.01nm）上的距离，以及激活过程的时间尺度（以微秒-纳秒为单位）。由于这种蛋白质通常位于脑细胞的细胞膜中，它需要细胞膜的一些脂质成分才能正常工作，我们也将研究这些成分中哪些是必需的。最后，如果时间允许，我们将确定被激活的受体如何将信号传递给信号分子链中的下一个组成部分，以进一步揭示这一重要的信号机制。来自这个前沿项目的所有信息将增加我们对大脑信号传输的总体理解，并有助于未来的药物设计和疾病控制。

Many of our neurological functions are controlled through receptor proteins (called GPCRs) residing in the brain, and for this reason it has been estimated that about 30% of all drugs we use act on these receptors, of which there are more than 800. The functions controlled by GPCRs are numerous, and one receptor may be involved in many responses. We need to find out how these receptors work, and it has been said that this is the major challenge of current structural biology (Lagerstrom & Schioth, 2008, Nature Reviews Drug Discovery, 7, 339-57). To help in that process of understanding, and as a result of recent breakthroughs, it is now possible to make a very limited number of GPCRs so that we can start to discover how they function. The one we have elected to work on is involved in pain management, neuromuscular diseases such as Parkinson s and the control of appetite, and hence obesity control. We have been able to express this mammalian brain receptor in simple E. coli bacterial cells, and it is functionally active to make any of the work we do relevant to its function in the brain. As with many of these receptors, they are activated by the binding of small molecules, and although we can monitor this binding, the important aspect is to understand and investigate how the protein is then activated and how it sends its signal to other proteins and then ultimately the cell. We will, therefore, express and purify the receptor, activate it with hormone, and then use state-of-the-art methods to probe the receptor changes during the activation process. These methods can be used to measure distances at the nanoscale (0.5-8nm +/- 0.01nm) in these nanodetectors , as well as the time scale (in microseconds - nanoseconds) for the activation process. Since this protein normally sits in a membrane of the brain cell, it needs some of the lipid components of the membrane to function properly, and we will also investigate which of these components are required. Finally, if time permits, we will determine how the activated receptor passes the signal to the next component in the chain of signalling molecules, to unravel a little more about this vital signalling mechanism. All the information from this cutting edge project will add to our general understanding of brain signal transmission, and help in future drug design and disease control.