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: 8129851
• 负责人: Robert E. Thorne
• 金额: $ 13.42万
• 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 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8129851
• 关键词: Acceleration; Affect; Applied Research; Area; Asia; Biotechnology; Calibration; Chemicals; Collaborations; Commit; Complex; Crystallization; Crystallography; Data Collection; Development; Drops; Europe; Evaluation; Film; Genes; Government; Growth; Harvest; Image; Image Analysis; Imaging technology; In Situ; Injection of therapeutic agent; Kinetics; Knowledge; Life; Liquid substance; Marketing; Mass Spectrum Analysis; Measurement; Membrane Proteins; Methods; Molds; Molecular; Molecular Medicine; Molecular Structure; Nucleic Acids; Optics; Organized by Structure Protein; Outcome; Pattern; Performance; Persons; Pharmacologic Substance; Phase; Positioning Attribute; Process; Production; Productivity; Property; Protein Structure Initiative; Proteins; Resources; Retrieval; Roentgen Rays; Sampling; Screening procedure; Series; Shapes; Shipping; Ships; Small Business Technology Transfer Research; Source; Speed; Spottings; Structure; Surface; Synchrotrons; Technology; Testing; United States National Institutes of Health; Universities; Virus; Work; X ray diffraction analysis; X-Ray Crystallography; X-Ray Diffraction; base; commercialization; cost; design; drug discovery; improved; insight; meetings; new technology; novel strategies; programs; protein expression; protein structure; prototype; response; structural genomics; success; three dimensional structure; tool

DESCRIPTION (provided by applicant): Our understanding of the molecular mechanisms of life is increasingly based upon our knowledge of the three dimensional structure of proteins, nucleic acids, viruses and biomolecular complexes. Structure provides insight into function (or malfunction), and provides a starting point for modern drug discovery and molecular medicine. Biomolecular structures are most often determined using X-ray crystallography of crystallized biomolecules. Over the last decade, high-throughput methods have been introduced that have automated many aspects of protein expression, purification, crystallization and crystallography, and that have lowered the cost per structure determined. However, the growth and harvesting of protein crystals remains a major bottleneck in the pipeline from gene to three-dimensional molecular structure and from structure to pharmaceutical therapy. This Phase II STTR proposal is focused on developing and commercializing improved methods for conventional and high-throughput crystallization and for crystal harvesting and X-ray data collection. An examination at Cornell University of how liquid contact lines interact with surfaces has led to a simple technology for precisely defining the positions of dispensed liquid drops and firmly holding them to those positions, regardless of their chemical composition. This technology forms the basis for a new approach to protein crystallization plates that eliminates the liquid-confining wells of conventional plates. These new plates promise to provide precise control over drop position and shape, resulting in more reproducible crystallization kinetics and simplified image analysis. They will allow hanging and sitting drop growth of soluble and membrane proteins using a single plate, and in situ optical, UV and X-ray analysis with low background. They will meet a critical need for plates optimized for easy X-ray examination of screening and crystallization outcomes and also allow in situ structure determination. They will be compatible with all existing drop dispensing and plate handling hardware, lowering barriers to market entry. This project will continue the scientific and commercial development of these plates and explore other biomedical applications of drop pinning technology. In Phase I we successfully developed and commercialized several new tools for crystal retrieval and X-ray data collection. We will continue this development in Phase II, focusing on improved tools for microcrystallography, for automated sample mounting, and for X-ray beam alignment and energy measurement. Together, these technologies should have significant impact on the productivity of high- throughput structural genomics and drug discovery efforts. PUBLIC HEALTH RELEVANCE: Our understanding of the molecular mechanisms of life is increasingly based upon our knowledge of the three dimensional structure of proteins, nucleic acids and viruses. Structure provides insight into function (or malfunction), and provides a starting point for modern drug discovery and molecular medicine. This project will develop and commercialize new technologies for use in determining the protein structures by X-ray crystallography that promise to speed up the progression from gene to pharmaceutical therapy.

描述（由申请人提供）：我们对生命分子机制的理解越来越多地基于我们对蛋白质，核酸，病毒和生物分子复合物的三维结构的了解。结构提供了有关功能（或故障）的洞察力，并为现代药物发现和分子医学提供了一个起点。生物分子结构最常使用结晶生物分子的X射线晶体学确定。在过去的十年中，已经引入了高通量方法，这些方法已经自动化了蛋白质表达，纯化，结晶和晶体学的许多方面，并降低了确定的每个结构的成本。然而，蛋白质晶体的生长和收获仍然是从基因到三维分子结构以及从结构到药物治疗的管道中的主要瓶颈。 该II期STTR提案的重点是开发和商业化改进的传统和高通量结晶的方法，以及用于晶体收集和X射线数据收集的方法。康奈尔大学（Cornell University）对液体接触线如何与表面相互作用的检查，导致了一项简单的技术，可以精确地定义分配的液滴液滴的位置并将其牢固地固定在这些位置上，无论其化学成分如何。该技术构成了一种新的蛋白质结晶板的基础，从而消除了常规板的液体填充井。这些新板有望提供对跌落位置和形状的精确控制，从而导致更可重现的结晶动力学和简化的图像分析。它们将允许使用单个板以及以低背景的原位光学，紫外线和X射线分析来悬挂可溶性和膜蛋白的降落和静置的滴落生长。他们将满足对优化板的临界需求，以轻松检查筛选和结晶结果，并允许原位结构确定。它们将与所有现有的Drop分配和盘子处理硬件兼容，从而降低了进入市场的障碍。该项目将继续对这些板的科学和商业开发，并探索滴钉技术的其他生物医学应用。 在第一阶段，我们成功地开发了几种用于晶体检索和X射线数据收集的新工具。我们将在第二阶段继续进行这种发展，重点是改进微晶学，自动样品安装以及X射线束对准和能量测量的工具。总之，这些技术应对高通量结构基因组学和药物发现工作的生产率产生重大影响。 公共卫生相关性：我们对生命的分子机制的理解越来越多地基于我们对蛋白质，核酸和病毒的三维结构的了解。结构提供了有关功能（或故障）的洞察力，并为现代药物发现和分子医学提供了一个起点。该项目将开发并商业化新技术，用于通过X射线晶体学确定蛋白质结构，该技术有望加快从基因到药物治疗的发展。