FUNDAMENTAL SCIENCE OF PROTEIN CRYSTALS FOR STRUCTURAL GENOMICS

结构基因组学蛋白质晶体基础科学

• 批准号: 8363567
• 负责人: Robert E. Thorne
• 金额: $ 3.37万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363567
• 关键词: Affect; Biological; Cool-X-A; Data Collection; Data Set; Disease; Funding; Glass; Grant; Light; Methods; Molecular Structure; National Center for Research Resources; Principal Investigator; Protein Dynamics; Proteins; Protocols documentation; Research; Research Infrastructure; Resources; Roentgen Rays; Science; Source; Structure; Temperature; United States National Institutes of Health; Work; base; cold temperature; cost; improved; macromolecule; protein structure; structural genomics; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. X-ray cryocrystallography is the primary tool used to determine the 3D molecular structure of proteins and other biological macromolecules, and to date roughly 50,000 unique structures having been determined. We are investigating several factors that affect the quality of the structural information that is obtained, and attempting to develop improved protocols. Specific questions being investigated include: Why does cooling protein crystals disorder them? How do X-rays damage the crystal, and how do low temperatures reduce this damage? How do the method by which a protein crystal is cooled and the cooling rate affect its structure? How does protein structure evolve with temperature, especially in the vicinity of the glass transition? Can variable temperature data collection shed new light on protein dynamics? Our planned work is based on two significant discoveries that have allowed us to successfully cool crystals at rates from 0.1 K/s to 100,000 K/s, and to successfully collect structural quality data sets at arbitrary temperatures between 300 K and 100 K, in both cases without the use of penetrating cryoprotectants.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 X射线冷冻同位素照相术是用于确定蛋白质和其他生物大分子的3D分子结构的主要工具，迄今为止已经确定了大约50，000个独特的结构。 我们正在研究影响所获得的结构信息质量的几个因素，并试图开发改进的协议。 正在研究的具体问题包括：为什么冷却蛋白质晶体会使它们紊乱？ X射线是如何破坏晶体的，低温又是如何减少这种破坏的？ 蛋白质晶体的冷却方法和冷却速度如何影响其结构？ 蛋白质结构如何随温度变化，特别是在玻璃化转变附近？可变温度数据收集可以为蛋白质动力学提供新的线索吗？ 我们计划的工作是基于两个重要的发现，使我们能够成功地冷却晶体的速率从0.1 K/s到100，000 K/s，并成功地收集结构质量数据集在任意温度之间的300 K和100 K，在这两种情况下，没有使用渗透冷冻保护剂。