Equipment to Support Protein Crystallisation in the York Structural Biology Laboratory

利用X射线结晶学测定生物大分子的结构提供了有关生物系统如何在单个分子水平上工作的信息。这些信息改变了我们对生命一些基本过程的理解。一个例子是蛋白质血红蛋白在有氧和无氧情况下的晶体结构，它解释了血细胞如何能够将氧气从肺部输送到组织中。第二个例子是如何调节糖原磷酸化酶等代谢酶的活性，使细胞根据有机体的营养状态燃烧或储存食物。此外，关键蛋白质的晶体结构可以提供有关疾病或感染在分子水平上发生的重要信息，并可以指导新药的开发。一个引人注目的和热门的例子是确定流感病毒神经氨酸酶的结构。这种结构被用来指导药物瑞乐沙和随后的达菲的开发。此外，神经氨酸酶和其他流感蛋白的结构让我们能够理解为什么流感的变种(如禽流感或1918年的西班牙流感)如此具有毒性，或许可以为开发更好的药物提供指导。在确定结构之前，蛋白质结晶学中有许多步骤。这个过程始于大量生产感兴趣的蛋白质。下一步，也是关键的一步，是生产蛋白质的晶体。这可能是一个漫长而困难的过程，要找到合适的溶液条件才能形成晶体。晶体是必要的，因为当你将X射线照射到晶体上时，你会获得一个衍射图，通过大量的努力，你可以从这个衍射图中提取出分子结构的图像。因此，成功提供晶体是X射线分析成功的基础和决定因素。近年来，在溶液条件的设计和可用于建立大量结晶试验的机器人设备方面都有不断的改进。一个特别重要的发展是使用了非常小的、纳米升大小的液滴。这减少了需要使用的蛋白质数量，增加了可以进行的结晶试验的次数，在某些情况下，小液滴增加了形成晶体的成功率。约克结构生物学实验室(YSBL)是欧洲最大的致力于蛋白质结构测定和分析的实验室之一。YSBL的科学家不仅在确定许多重要蛋白质的结构方面做出了重大贡献，而且在发展X射线晶体结构确定所需的实验和计算方法方面也做出了重大贡献。这种方法开发的一个要素是设计新的结晶屏解决方案，并与制造商合作开发改进的机器人设备。该应用程序用于支持YSBL结晶试验的机器人设备的升级。这将使约克的科学家能够受益于设备的一些进步，更快地确定更多蛋白质的结构，但也可以继续与制造商合作，进行进一步的改进。

The determination of the structure of biological macromolecules using X-ray crystallography is providing information about how biological systems work at the level of individual molecules. This information has transformed our understanding of some of the fundamental processes of life. One example is the crystal structures of the protein haemoglobin in the presence and absence of oxygen which explain how blood cells are able to transport oxygen from the lungs to the tissues. A second example would be how the activity of metabolic enzymes such as glycogen phosphorylase are regulated so that cells either burn or store food depending on the nutritional state of the organism. In addition, crystal structures of key proteins can give important information about what happens at the molecular level in disease or infection and can guide the development of new drugs. A striking and topical example is the determination of the structure of the enzyme neuraminidase from the influenza virus. The structure was used to direct the development of the drugs relenza and subsequently tamiflu. Moreover, structures of neuraminidase and other influenza proteins allow us to understand why variants of flu (such as avian influenza or the Spanish flu of 1918) are so virulent, perhaps providing guidance on developing even better drugs. There are a number of steps in protein crystallography before a structure can be determined. The process starts with the production of large quantities of the protein of interest. The next, key step is to produce crystals of the protein. This can be a long and difficult process, finding the right solution conditions under which crystals will form. Crystals are necessary as when you shine X-rays on a crystal, you obtain a diffraction pattern from which, with a lot of effort, you can extract an image of what the structure of the molecule looks like. Therefore, the success of X-ray analysis is underpinned and determined by successful provision of crystals. In recent years, there have been continual improvements in both the design of the solution conditions and the robotics equipment available for setting up large numbers of crystallisation trials. A particularly important development has been the use of very small, nano-litre sized drops. These reduce the amount of protein that needs to be used, increases the number of crystallisation trials that can be conducted and in some cases, the small drops have increased the success of forming crystals. The Structural Biology Laboratory at York (YSBL) is one of the largest laboratories in Europe dedicated to the determination and analysis of protein structure. Scientists in YSBL have made major contributions by not only determining the structures of many important proteins, but also in developing the experimental and computational methods that are required for X-ray crystal structure determination. One element of this methods development has been devising new crystallisation screen solutions and also working with the manufacturers in developing improved robotics equipment. This application is for an upgrade to the robotics equipment that supports crystallisation trials in YSBL. This will allow scientists at York to benefit from some of the advances in equipment, to determine the structures of more proteins, more rapidly, but also to continue to work with manufacturers in making further improvements.