Analytical Ultracentrifugation for Complex Systems

复杂系统的分析超速离心

• 批准号: 6520441
• 负责人: THOMAS M LAUE
• 金额: $ 21.93万
• 依托单位: UNIVERSITY OF NEW HAMPSHIRE
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6520441
• 关键词: DNA damage; analytical method; analytical ultracentrifugation; bioluminescence; biosensor device; chemical binding; fluorescence; intermolecular interaction; molecular dynamics; radiotracer; stereochemistry; technology /technique development

APPLICANT'S DESCRIPTION: The characterization of the interactions between molecules is central to our understanding of both normal and pathological biochemistry. Consequently, the accurate description of molecular interactions plays a critical role in the development of safe and effective drugs. Improvements in the instruments and methods for characterizing molecular interactions are necessary for researchers to efficiently advance basic medical science. Analytical ultracentrifugation is one of the most rigorous, powerful and adaptable methods for determining the strength, stoichiometry and size of molecular complexes, but is currently limited to the analysis of relatively simple biochemical systems. With further development, analytical ultracentrifugation would be applicable to problems that are far more complex than can be investigated using current technology. The overall goal is to bring the power and rigor of analytical ultracentrifugation to bear on problems of interest in molecular and cellular biology. Collaborative efforts have been established with several NIH-funded researchers who use analytical ultracentrifugation in their work and who require the advances provided by the three specific aims of this proposal: 1) Development of rapid-scan absorbance optics. A photodiode-array-based detector capable of providing concentration profiles in < 1 s and at a 10 um radial resolution will be developed. Wavelength selection will require < 1 S with a reproducibility better than +/- 1 nm. These improvements will allow the analysis of rapidly-sedimenting boundaries at multiple wavelengths. 2) Development of radiolabel and Immuno-chemiluminescence detectors. Cell windows and centerpieces will be developed to permit the use of a luminescence detector for radiolabel and immuno-chemiluminescence detection. The specificity and sensitivity will allow the characterization of molecules under conditions approaching those found in vivo. The new centerpieces will decrease sample volume by up to 40-fold, while increasing the number of samples per experiment up to 4-fold, making it possible to rapidly and effectively conduct experimental protocols involving a large number of variables. 3) Development of an integrated ultracentrifuge operating system. A new instrument operating system will be developed to make the instrument easier to operate and improve the efficiency of data collection and analysis by logically organizing data acquisition procedures.

申请人的描述： 分子是我们理解正常和病理的核心 生物化学因此，分子相互作用的准确描述 在开发安全有效的药物方面起着关键作用。 表征分子的仪器和方法的改进 互动是研究人员有效推进基础医学的必要条件。 科学分析超离心是一种最严格，最有力的 以及确定强度、化学计量和尺寸的适应性方法 分子复合物，但目前仅限于分析相对 简单的生化系统随着进一步发展，分析 超离心法将适用于更为复杂的问题 比现有技术所能研究的要多总体目标是 分析超集中的力量和严谨性， 对分子和细胞生物学感兴趣。合作努力已 与几位NIH资助的研究人员建立了关系， 他们在工作中的极端集中，谁需要提供的进步， 该提案的三个具体目标： 1)快速扫描吸收光学的发展。基于光电二极管阵列的 能够在< 1 s内提供浓度分布和10 μ m 将开发径向分辨率。波长选择需要< 1 S 再现性优于+/-1nm。这些改进将使 在多个波长下分析快速沉积边界。 2)放射性标记和免疫化学发光检测器的研制。细胞 窗户和中心装饰将被开发，以允许使用发光 用于放射性标记和免疫化学发光检测的检测器。特异性 和灵敏度将允许在一定条件下表征分子 接近那些在体内发现的。新的中心装饰将减少样品 体积高达40倍，同时增加每个实验的样品数量 高达4倍，使其能够快速有效地进行 涉及大量变量的实验方案。 3)集成超高速操作系统的开发。一个新 将开发仪器操作系统，使仪器更容易 操作和提高数据收集和分析的效率， 组织数据采集程序。