Tools for High Throughput Protein Crystallization and Structure Determination

描述(由申请者提供)：项目摘要：美国国立卫生研究院承诺投入大量资源用于传统的和通过其蛋白质结构倡议高通量方法确定蛋白质结构，欧洲和亚洲也做出了类似的承诺。对可溶性蛋白质研究结果的调查表明，主要的瓶颈是获得高质量的晶体和获得准确结构的高质量的衍射数据。MiTeGen，LLC已成功地将康奈尔大学的学术发现迅速转化为对X射线衍射管道产生重大影响的商业产品。该项目的第一阶段将专注于基于流行的自动滴注技术开发新的结晶平台。康奈尔大学正在进行的基础和应用研究正在研究蛋白质晶体生长的物理学，以及含有蛋白质的液滴的分配、动力学和限制。这些研究已经从根本上导致了结晶板的新设计，这些设计承诺减少存储需求，允许对晶体质量进行原位X射线检查，改善平衡动力学和结晶实验的重复性，并提高提取晶体进行X射线结构测定的易用性。同时，将开发用于晶体检索和X射线结构确定的工具，这些工具与这些结晶平台集成在一起，适合于“远程”和自动化的晶体安装。与现有技术相比，这些板材和工具设计具有极强的成本竞争力。第一阶段的目标是优化这些设计，并开始商业原型生产。与工业和学术自动化团体(特别是与PSI相关的团体)的密切合作将确保设计是可行的。第二阶段将专注于更先进的设计，纳入康奈尔大学基础和应用研究的见解，自动化晶体检索，以及X射线显微镜支架的开发。相关性：蛋白质结晶学是现代结构基因组学和药物发现工作的核心组成部分。通过促进更快、更有效的蛋白质结构确定，拟议的研究将有助于了解疾病的潜在机制，并开发新的治疗方法。

DESCRIPTION (provided by applicant): Project Summary: The NIH has committed substantial resources to both traditional and-through its Protein Structure Initiative-high-throughput approaches to protein structure determination, and similar commitments have been made in Europe and Asia. A survey of the results for soluble proteins shows that the main bottlenecks are obtaining high-quality crystals and obtaining high-quality diffraction data for accurate structures. Mitegen, LLC has succeeded in rapidly turning academic discoveries at Cornell into commercial products that are having a major impact on the X-ray diffraction pipeline. Phase I of this project will focus on development of new crystallization platforms based on popular automated drop dispensing technology. Ongoing fundamental and applied studies at Cornell are investigating the physics of protein crystal growth and of the dispensing, dynamics and confinement of protein-containing liquid drops. These studies have led to fundamentally new designs for crystallization plates that promise to reduce storage requirements, allow in-situ X-ray examination of crystal quality, improve equilibration kinetics and the reproducibility of crystallization experiments, and improve the ease with which crystals can be retrieved for X-ray structure determination. Concurrently, tools will be developed for crystal retrieval and X-ray structure determination that integrate with these crystallization platforms and that are suitable for "remote" and automated crystal mounting. These plate and tool designs promise to be highly cost competitive with existing technologies. The Phase I goal is to optimize these designs and begin commercial prototype production. Close collaboration with industrial and academic automation groups (particularly those associated with the PSI) will ensure that the designs are viable. Phase II will focus on more advanced designs incorporating insights from fundamental and applied studies at Cornell, on automated crystal retrieval, and on development of mounts for X-ray microscopies. Relevance: Protein crystallography is a central component of modern structural genomics and drug discovery efforts. By facilitating faster and more efficient protein structure determination, the proposed research will assist in understanding the mechanisms underlying disease and in developing new treatments.