'Gel-to-Grid' cryoEM of membrane proteins using SMALP technology

自从显微镜在17世纪中叶问世以来，生物学家一直惊叹于那些往往比人类头发的宽度小许多倍的结构是如何支撑着生命的奇迹的。了解这些微小但形成精致的粒子的结构使我们能够揭开大自然的秘密。反过来，这些洞察力使我们在理解人类和动物疾病方面取得了一些最重要的进展。例如，我们对病毒结构的了解使我们能够开发更有效的疫苗，而蛋白质结构则彻底改变了我们开发药物的速度。了解这些纳米级的结构还可以让我们回答一些最重要的科学问题，从遗传物质如何复制到肌肉是如何工作的。这些洞察力得益于用于可视化这些小颗粒的方法的定期突破。每一次突破都能让人们更快地看到越来越多具有挑战性的样本的更多细节。想想看，在300年的时间里，我们已经能够从可视化细胞发展到科学家可以例行公事地看到组成细胞的单个原子的排列。在伯明翰和华威的实验室里，我们一直在研究一种方法，这种方法将使我们能够可视化生物学中一些最重要的蛋白质的原子排列。这些蛋白质生活在细胞周围的膜中，执行许多使细胞存活的基本工作。它们允许营养物质进入细胞，并允许废物离开，它们还调节信号的进出细胞。这些功能使研究人员对它们格外感兴趣，但或许更重要的是，它们也是开发动物和人类健康新疗法的最重要目标。不幸的是，在过去的50年里，这些蛋白质被证明是非常难以研究的，并使大量科学家感到困惑。2009年，我们发现了一种制造这些蛋白质的全新方法，它使用了一种更常见于汽车仪表板和油漆中的简单聚合物。这种方法将蛋白质“包裹”在聚合物“带”中，使其保持稳定，从而使我们能够对其进行研究。在这个项目中，我们将开发一种新的方法，使这些聚合物带蛋白更容易转移到显微镜中，以便对它们进行成像。我们希望该方法将使这一过程比目前的方法容易得多，并将允许拍摄更多的重要膜蛋白的图像。这将使我们对生物世界有一个更完整的“原子图景”，这将极大地帮助新疗法的发展。

Since the development of the microscope in the middle of the 17th century biologists have marvelled at how structures that are often many times smaller than the width of a human hair underpin the marvel of life. Understanding the structures of these tiny yet exquisitely formed particles has allowed us to unlock the secrets of nature. In turn these insights have allowed us to make some of the most important advances in our understanding of disease in humans and animals. For example, our understanding of the structure of viruses has allowed us to develop more effective vaccines while structures of proteins have revolutionised the speed with which we develop drugs. Understanding these nano-scale structures also allows us to answer some of the most important questions in science from how genetic material replicated to how muscles work. These insights have been enabled by regular breakthroughs in the methods used to visualise these small particles. Each breakthrough allows more detail to be seen more rapidly for more and more challenging samples. Just consider, in 300 years we have been able to move from visualising cells to a position where scientists can routinely see the arrangement of individual atoms that make up the cells. In our laboratories in Birmingham and Warwick we have been working on a method that will allow us to visualise the atomic arrangement of some of the most important proteins in biology. These proteins live in the membrane that surrounds cells and carry out some many essential jobs that allow cells to survive. They allow nutrients into the cell and allow waste products to leave, they also mediate the movement of signals into and out of the cell. These functions make them exceptionally interesting to researchers, but perhaps more importantly they are also the most important targets for developing new therapeutics for animal and human health. Unfortunately these proteins have proved to be exceptionally difficult to study and have confounded a large number of scientists over the past 50 years. In 2009 we found an entirely new way of making these proteins by using a simple polymer more commonly found in car dashboards and paint. This method "wraps" the protein in a polymer "belt" that keeps it stable allowing us to study it. In this project we am to develop a new method that will make it easier to transfer these polymer belted proteins into microscopes to allow them to be imaged. We hope that the method will make this process much easier than current methods and will allow more images to be taken of important membrane proteins. This should allow us the make a more complete "atomic picture" of the biological world greatly aiding the development of new therapeutics.