X-ray Diffraction Equipment for Macromolecular Crystallography at York

了解分子结构，即分子的三维形状，通常是了解分子如何发挥作用的关键。对于协调维持活细胞的生化过程的大型生物分子来说尤其如此。确定蛋白质结构的一个重要方法是首先培养这些蛋白质的晶体，然后用x射线衍射法记录它们对x射线的散射。通过收集不同晶体方向的一系列x射线图像，我们积累了由10,000 - 500,000个“观测”组成的三维数据集。使用计算方法，我们可以使用这些数据来“解决”（定义原子的位置）蛋白质在晶体中存在的结构。由于其对蛋白质作用的生物学机制和蛋白质在生物技术和生物医学应用中的开发的广泛兴趣，结构生物学实验室每年产生1000个蛋白质晶体。在这里，我们请求资金更新我们的x射线衍射设备套件，包括x射线发生器，一个测量晶体x射线衍射的探测器，以及一个可以自动和连续地将晶体放入x射线束的机械臂。使用这个仪器，我们每年可以测试大约3000个晶体。我们拒绝质量不足的晶体进行结构测定，我们优先考虑最好的晶体（~1000）进行数据收集。这些晶体被送到同步加速器，钻石光源（DLS），这是一个产生超强x射线束的国家设施。DLS可以快速准确地获得高分辨率的衍射数据。DLS的波束时间是一种宝贵而有限的资源，对波束时间的需求超过了波束的可用性。我们实验室平均每月轮班一次。因此，在我们自己的实验室中进行晶体的预筛选对于有效利用DLS资源至关重要，以便将光束时间用于收集有用的数据而不是晶体测试。实验室的生产力可以通过向一个名为蛋白质数据库的公共数据库提交约700个晶体结构以及在同行评审期刊上发表310篇描述这些结构的论文来证明。我们确定的这些结构支撑着改进洗衣粉的开发、生物燃料的生物质转化科学和新药的开发。

Understanding molecular structure, that is the 3D shape of a molecule, is often the key to understanding how that molecule performs its function. This is particularly true for large biological molecules that orchestrate the biochemical processes that maintain living cells. An important method of determining the structure of proteins is first to grow crystals of these proteins and then to record their scattering of X-rays by what is termed X-ray diffraction. By collecting a series of X-ray images at different crystal orientations, we accumulate three-dimensional data sets consisting of 10,000 - 500,000 'observations'. Using computational methods we can then use these data to 'solve' (define the positions of atoms) the structure of the protein as it exists in the crystal. With its broad interest in the biological mechanisms of protein action and the exploitation of proteins for biotechnological and biomedical applications, the Structural Biology Laboratory generates 1000s of protein crystals a year. Here we are requesting funds to renew our suite of X-ray diffraction equipment including an X-ray generator, a detector for measuring X-ray diffraction by crystals, and a robot arm that can automatically and serially place crystals into the X-ray beam. Using this apparatus, we can test approximately 3000 crystals each year. We reject crystals that are of insufficient quality for structure determination and we prioritise the best crystals (~1000) for data collection. These crystals are sent to a synchrotron, the DIAMOND Light Source (DLS), a national facility where super-intense X-ray beams are generated. At DLS we can obtain high resolution diffraction data rapidly and accurately. Beam time at DLS is a valuable and limited resource and demand for beam time exceeds beam availability. Our laboratory has one shift every month on average. For this reason, the pre-screening of crystals in our own laboratory is essential for the efficient use of the DLS resource, so that the beam time is used to collect useful data rather than crystal testing. The productivity of the laboratory is evidenced by the submission of ~700 crystal structures into a publicly accessible database, called the Protein Data Bank, and the publication in peer-reviewed journals of 310 papers describing these structures. The structures we have determined are underpinning the development of improved washing powders, the science of biomass conversion for biofuels and the development of new medicines.