Acquisition of Automated Nanoscale Crystallization Equipment

描述（由申请人提供）：在这份提案中，麻省理工学院生物系和化学系的六名教员要求为安装在麻省理工学院结构生物学核心设施的自动化大分子结晶设备提供资金。直到今年，麻省理工学院的结构生物学家还缺乏自动化的纳米级结晶设备，而这种设备在大分子晶体学设施中越来越普遍。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的。这种实验方法是费力的，重复的，浪费的-消耗大量的样品是昂贵的生产。在结构蛋白质组学项目的推动下，最近的技术发展已经使设备在筛选和初始优化方面很大程度上取代了人工方法。作为第一步，这组结构生物学家最近购买了一个用于纳米级结晶筛选的自动液体处理系统。前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，并且已经启动，改进的再现性使重点优化策略成为可能，并且在创纪录的时间内发现了初始命中点。所申请的资金将用于扩大和补充该液体处理系统，为最新的结晶设施提供必要的设备，即，一个移液机器人，以组成精确定制的网格屏幕，一个组合成像和存储系统，以跟踪结晶试验，以及一个存储和网络服务器，具有足够的容量，记录并向研究人员展示结晶实验的预定成像结果。这个综合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标准进行系统的结晶实验。这将使纯化样品数量有限的项目变得可行——减少10-15倍的蛋白质使用是一个关键的好处，因为我们的许多目标是大分子复合物或需要在真核细胞中表达的蛋白质。最后，该系统将在其他方面节省资金，特别是在复杂的大分子晶体学项目上，节省工资时间和消耗品。公共卫生相关性：主要用户的研究兴趣跨越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系统、更有效地进行结构项目。它将使我们能够追求当前最感兴趣的生物和生物医学结构目标，巧合的是，这些目标往往是最难生产足够高纯度样品材料的目标。

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.