New Approaches for Protein Crystallization, Cryopreservation and Radiation Damage

蛋白质结晶、冷冻保存和辐射损伤的新方法

• 批准号: 7580962
• 负责人: Robert E. Thorne
• 金额: $ 25.77万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-15 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7580962
• 关键词: Affect; Age; Asia; Behavior; Biological; Commit; Cryopreservation; Crystallization; Crystallography; Data; Data Collection; Disease; Drops; Europe; Goals; Growth; Image; Kinetics; Maps; Medicine; Methods; Microfabrication; Molecular Structure; Outcome; Phase; Process; Property; Protein Structure Initiative; Proteins; Protocols documentation; Radiation; Research; Resolution; Resources; Roentgen Rays; Science; Solutions; Solvents; Speed; Structural Biologist; Structure; Surface; Surveys; Synchrotrons; Techniques; Technology; United States National Institutes of Health; Viral Proteins; Vision; Work; X ray diffraction analysis; X-Ray Diffraction; aqueous; base; coping; drug discovery; improved; interest; new technology; novel strategies; protein structure; structural genomics

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 X-ray diffraction data for accurate structures. The long-term goals of this project are to understand the underlying science involved in macromolecular crystal growth, in cryopreservation of protein crystals, and in radiation damage during X-ray data collection, and to use this understanding to develop practical protocols and technologies. These studies utilize a wide array of physical methods, from synchrotron-based X-ray diffraction and imaging to microfabrication. Our specific goals are: (1) To understand the fundamental processes in cryopreservation, and to develop more effective methods to capture and preserve protein and crystal structure based on ultra- fast cooling, the kinetic phase diagrams of aqueous mixtures, and the thermal expansion behavior of solvent and crystal; (2) To. develop methods for cryopreservation and long-term storage of protein solutions that preserve crystallization behavior; (3) To quantify and understand radiation damage in protein crystals, and to develop protocols for reducing this damage in small crystals; and (4) To explore protein drop-surface interactions and drop dynamics relevant to protein crystallization. Relevance: Protein crystallography is a central component of modern structural genomics and drug discovery efforts, which promise to revolutionize medicine in the 21st century. The proposed research will facilitate more rapid and accurate determination of protein structures, and thereby 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已为传统和 - 通过其蛋白质结构倡议提供了大量资源 - 蛋白质结构确定的高通量方法，并且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，并获得了高质量的X射线衍射数据以进行准确的结构。该项目的长期目标是了解与大分子晶体生长，蛋白质晶体冷冻保存以及X射线数据收集过程中的放射线损害有关的基本科学，并利用这种理解来开发实用的方案和技术。这些研究利用了各种物理方法，从基于同步加速器的X射线衍射和成像到微分化。我们的具体目标是：（1）了解冷冻保存中的基本过程，并开发基于超快速冷却的蛋白质和晶体结构的更有效的方法，水性混合物的动力学相图以及溶剂和晶体的热膨胀行为； （2）至。开发用于保存结晶行为的蛋白质溶液的冷冻保存和长期存储的方法； （3）量化和理解蛋白质晶体中的辐射损伤，并制定用于减少小晶体损伤的方案； （4）探索与蛋白质结晶相关的蛋白质下降表面相互作用和滴动力学。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，该工作有望在21世纪彻底改变医学。拟议的研究将促进更快，更准确地确定蛋白质结构，从而有助于理解疾病潜在的机制和开发新疗法。