Tools for High Throughput Protein Crystallization and Structure Determination

高通量蛋白质结晶和结构测定工具

• 批准号: 7109592
• 负责人: ROBERT E THORNE
• 金额: $ 10万
• 依托单位: MITEGEN, LLC
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-23 至 2010-09-22
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7109592
• 关键词: X ray crystallography; bioengineering /biomedical engineering; biomedical automation; biophysics; chemical kinetics; crystallization; functional /structural genomics; high throughput technology; method development; protein structure function; telemetry

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.

描述（由申请人提供）：项目摘要：NIH为传统和直通蛋白质结构的蛋白质结构倡议指定蛋白质结构的确定方法提供了大量资源，并在欧洲和亚洲也做出了类似的承诺。对可溶性蛋白的结果的调查表明，主要瓶颈正在获得高质量的晶体，并获得了准确结构的高质量衍射数据。 Mitegen，LLC成功地将康奈尔大学的学术发现迅速转变为对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重点介绍基于流行的自动滴分配技术的新结晶平台的开发。康奈尔大学正在进行的基本研究和应用研究正在研究蛋白质晶体生长的物理学以及含蛋白质液滴的分配，动力学和限制。这些研究从根本上为结晶板提供了新的设计，这些设计有望减少存储需求，允许对晶体质量的原位检查，改善平衡动力学以及结晶实验的可重复性，并提高晶体以确定X射线结构的易感性。同时，将开发工具用于与这些结晶平台集成的晶体检索和X射线结构的确定，并且适用于“远程”和自动晶体安装。这些板和工具设计有望与现有技术具有高度的成本竞争力。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小组（尤其是与PSI相关的人）的密切合作将确保设计可行。第二阶段将着重于更先进的设计，这些设计结合了康奈尔大学基础和应用研究，自动晶体检索的见解，以及用于X射线显微镜的坐骑的开发。相关性：蛋白质晶体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理解疾病潜在的机制和开发新疗法。