New Approaches to Radiation Damage and Cryoprotection

辐射损伤和冷冻防护的新方法

• 批准号: 6932448
• 负责人: ROBERT E THORNE
• 金额: $ 17.74万
• 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 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6932448
• 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现代生物过程和疾病机制理解以及新药设计的核心。x射线本身和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分辨率和精度——这对机械研究尤其重要——在不利的情况下，可能会阻止结构确定。强大的衍射和实时空间成像技术最近被用于解剖大分子晶体中的无序，并为理解这种损伤提供了新的工具。