Acquisition of Automated Nanoscale Crystallization Equipment

描述(由申请人提供)：在这份提案中，麻省理工学院生物和化学系的六名教员申请拨款，在麻省理工学院的结构生物学核心设施安装自动化的大分子结晶设备。直到今年，麻省理工学院的结构生物学家一直缺乏在大分子结晶学设施中变得普遍的自动化、纳米级结晶设备。通过人工移液进行了结晶筛选和优化。随后的观察也是手工进行的。这种实验方法费力、重复且浪费--耗费大量生产成本高昂的样品。在结构蛋白质组学项目的推动下，最近的技术发展已经提供了设备，这些设备现在在很大程度上取代了人工方法在筛选和初步优化方面的作用。作为第一步，这组结构生物学家最近购买了一种用于纳米级结晶筛选的自动液体处理系统。头两个月的结果超出了预期：样品材料有限的项目已经成为可能并已启动，重现性的提高使重点优化战略成为可能，并在创纪录的时间内找到了初步的成功。申请的资金将用于扩大和补充这一液体处理系统，为最新的结晶设施提供必要的设备--即一个准确组成定制网格屏幕的移液机器人，一个跟踪结晶试验的综合成像和存储系统，以及一个有足够容量记录和向调查人员展示预定结晶实验成像结果的存储和网络服务器。这一集成系统将极大地促进麻省理工学院的结构生物学。它将允许学生和博士后科学家按照今天的S标准系统地进行结晶实验。它将使提纯样品的数量受到限制的可行项目--蛋白质使用量减少10-15倍是一个关键好处，因为我们的许多目标是大分子复合体或需要在真核细胞中表达的蛋白质。最后，该系统将在其他地方节省资金，在受薪时间和消耗品上花费，特别是在困难的大分子结晶学项目上。公共卫生相关性：主要使用者的研究兴趣横跨现代生物学和生物化学的广度。拟议的设备将使我们能够更有系统地开展结构性项目，从而更有效率。它将使我们能够追求那些最新的生物和生物医学兴趣的结构目标，巧合的是，这些结构目标往往是最难生产足够高纯度样品材料的。

DESCRIPTION (provided by applicant): In this proposal, six faculty of the Departments of Biology and of Chemistry at MIT request funding for automated macromolecular crystallization equipment to be installed at MIT's Structural Biology Core Facility. MIT structural biologists have lacked, until this year, any of the automated, nanoscale crystallization equipment that is becoming common at macromolecular crystallography facilities. Crystallization screens and optimization have been performed by manually pipetting. Subsequent observation is manual as well. This experimental approach is laborious, repetitive, and wasteful - consuming large amounts of samples that are expensive to produce. Recent technology developments, driven by structural proteomics projects, have made available equipment that now largely supplants manual methods in screening and initial optimization. As a first step, this group of structural biologists recently purchased an automated liquid handling system for nanoscale crystallization screening. The results in the first two months have exceeded expectations: projects where sample material was limiting have become possible and have been initiated, improved reproducibility has enabled focused optimization strategies and initial hits have been found in record time. The requested funds will be used to expand and complement this liquid handling system, providing the equipment necessary to an up-to-date crystallization facility - namely, a pipetting robot to compose accurately customized grid screens, a combined imaging and storage system to follow crystallization trials, and a storage and web server with sufficient capacity to record and present to investigators the results of scheduled imaging of crystallization experiments. This integrated system will promote immeasurably structural biology at MIT. It will allow students and postdoctoral scientists to perform systematically crystallization experiments according to today<s standards. It will make feasible projects for which the amount of purified sample is limiting -- 10-15 fold reduction in protein used is a key benefit, because many of our targets are macromolecular complexes or proteins requiring expression in eukaryotic cells. Lastly, the system will save funds elsewhere, in salaried time spent and consumables expended, especially on difficult macromolecular crystallography projects. PUBLIC HEALTH RELEVANCE: The research interests of the primary users span the breadth of modern biology and biochemistry. The proposed equipment will allow us to pursue structural projects more systematically and thus more efficiently. It will enable us to pursue those structural targets of the most current biological and biomedical interest, which are, coincidentally, often the ones for which sufficient highly purified sample material is the hardest to produce.