A MOLECULAR RULER FOR DIRECT MEASUREMENT OF DISTANCE DISTRIBUTIONS IN SOLUTIONS

用于直接测量溶液中距离分布的分子尺

• 批准号: 7597961
• 负责人: PEHR A HARBURY
• 金额: $ 0.1万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-03-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7597961
• 关键词: Computer Retrieval of Information on Scientific Projects Database; DNA; Dyes; Event; Fluorescence Resonance Energy Transfer; Foundations; Funding; Grant; Institution; Knowledge; Label; Magnetic Resonance; Measurement; Molecular; Research; Research Personnel; Resources; Roentgen Rays; Shapes; Single-Stranded DNA; Site; Solutions; Source; Structure; Techniques; Testing; Time; United States National Institutes of Health; apomyoglobin; insight; molecular dynamics; nanoGold; protein folding; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To gain insight into the molecular events that underlie protein folding, knowledge of the tertiary structure of folding intermediates and molten globules would be extremely informative. However, conventional crystallographic and magnetic resonance techniques cannot characterize the dynamic, three-dimensional conformational ensembles that compose these states. While time-resolved fluorescence resonance energy transfer (FRET) studies can give some information on distance distributions, such analyses rely on assumptions about FRET dye orientations and the shapes of the distributions (generally Gaussian). We propose to use small-angle X-ray scattering in conjunction with heavy-atom replacement (SAXS-HR) to determine directly intramolecular distance distributions in heavy-atom duster labeled biomolecules. The results will constrain molecular-dynamics simulations to provide three-dimensional pictures of protein folding intermediates and molten globules. We propose to perform a proof of principle' test of the SAXS-HR technique: measurements on DNA duplexes and flexible DNA single strands labeled with nanogold clusters will demonstrate that site-specific distance distributions can be recovered by SAXS-HR. These experiments will provide a foundation for studies of the tertiary structure present in the apomyoglobin molten globule folding intermediate.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 为了深入了解蛋白质折叠的分子事件，折叠中间体和熔融球的三级结构的知识将是非常有益的。然而，传统的晶体学和磁共振技术不能表征的动态，三维构象合奏，组成这些国家。虽然时间分辨荧光共振能量转移（FRET）的研究可以提供一些信息的距离分布，这种分析依赖于假设的FRET染料的方向和分布的形状（一般是高斯）。我们建议使用小角X射线散射结合重原子置换（SAXS-HR）直接确定重原子簇标记的生物分子的分子内距离分布。这些结果将限制分子动力学模拟，以提供蛋白质折叠中间体和熔融球的三维图像。我们建议进行SAXS-HR技术的原理验证测试：对用纳米金簇标记的DNA双链体和柔性DNA单链的测量将表明，位点特异性距离分布可以通过SAXS-HR恢复。这些实验将为研究脱辅基肌红蛋白熔融球折叠中间体中存在的三级结构提供基础。