New Approaches to Radiation Damage and Cryoprotection

辐射损伤和冷冻防护的新方法

• 批准号: 6513897
• 负责人: ROBERT E THORNE
• 金额: $ 20.01万
• 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 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6513897
• 关键词: X ray crystallography; atomic force microscopy; cell water; conformation; cryoscience; cytoprotection; data collection methodology /evaluation; macromolecule; method development; oxygen; radiobiology; solvents; structural biology; thermostability

DESCRIPTION (provided by applicant): Macromolecular structures determined by X-ray crystallography are central to modern understanding of biological processes and disease mechanisms and to the design of new medicines. Crystal damage caused by the X-rays themselves and by the cryoprotective procedures used to minimize radiation damage reduces structure resolution and accuracy -- of particular importance to mechanistic studies -- and in unfavorable cases can prevent structure determination. Powerful diffraction and real-space imaging techniques have recently been used to dissect disorder in macromolecular crystals, and provide new tools for understanding this damage. The long term goals of this project are to characterize the systematics of radiation and cryoprotection-induced disorder using simple model proteins and viruses; to develop new procedures for reducing this disorder so that the full potential of high brilliance synchrotron sources for data collection can be exploited; and to apply these to selected systems of significant biological interest. Radiation damage and damage caused by flash cooling are complex phenomena, and no single factor is likely to produce dramatic improvements. Systematic experiments using X-ray diffraction and imaging, atomic force microscopy and other techniques will evaluate the most important factors, and explore how they may be combined to optimize overall data quality. This approach has significant and realistic potential to impact how macromolecular structures are determined, and thereby to contribute to their use in medical, pharmaceutical, and agricultural applications.

描述（由申请人提供）：由X射线晶体学确定的大分子结构是对生物过程和疾病机制的现代理解以及新药物设计的核心。由X射线本身以及用于最小化辐射损伤的冷冻保护程序造成的晶体损伤会降低结构的分辨率和准确性 - 特别重要 - 在不利的情况下，可以防止结构确定。最近已使用强大的衍射和真实空间成像技术在大分子晶体中解剖障碍，并提供了理解这种损害的新工具。 该项目的长期目标是使用简单的模型蛋白和病毒来表征辐射和冷冻保护诱导的疾病的系统。为了制定减少这种疾病的新程序，以便可以利用高光彩同步源的全部潜力；并将其应用于具有重大生物学意义的选定系统。辐射损伤和闪光冷却引起的损害是复杂的现象，没有任何一个因素可能会产生巨大的改进。使用X射线衍射和成像，原子力显微镜和其他技术的系统实验将评估最重要的因素，并探讨如何合并它们以优化整体数据质量。这种方法具有影响大分子结构的确定方式，从而有助于其在医疗，药物和农业应用中的使用。