USING WAXS TO STUDY MOLTEN GLOBULE STATES OF PROTEINS

使用蜡研究蛋白质的熔球态

• 批准号: 7722758
• 负责人: LEE MAKOWSKI
• 金额: $ 1.26万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7722758
• 关键词: Breathing; Chicago; Computer Retrieval of Information on Scientific Projects Database; Crystallography; Data; Funding; Grant; Institution; KLK3 gene; Lactalbumin; Life; Literature; Methods; Nature; Proteins; Quaternary Protein Structure; Research; Research Personnel; Resources; Solutions; Source; Structure; System; Time; Ubiquitin; United States National Institutes of Health; Universities; apomyoglobin; protein folding

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. Previous studies by our group at the APS have demonstrated that wide angle x-ray scattering (WAXS) from proteins in solution is highly sensitive to changes in secondary, tertiary and quaternary protein structure. It is particularly powerful when applied to dynamic or flexible systems such as partially folded proteins or ensembles of structure states which are not usually amenable to x-ray crystallography. Although there is a considerable body of literature on the transition of proteins from the native to the molten globule to the unfolded states, there still exists a considerable lack of understanding of what and how much structure is present in the intermediate molten globule state. This proposal seeks to use WAXS analysis of native, molten globule and denatured states of well-characterized proteins to elucidate the nature of the molten globule state. WAXS data will be collected from proteins such as apomyoglobin; lactalbumin, ubiquitin and lactoglobulin which have been induced to undergo molten globule formation and denaturation. Using methods of analysis developed in our studies of protein breathing, we will characterize the dynamics of the molten globule state of each of these proteins. These results will be compared to theoretical structure ensemble predictions performed by T. Sosnick at the University of Chicago. These studies will be followed by time-resolved WAXS studies of the more stable (in terms of life-time) intermediates within the protein group studied in order to follow both the changes in structure and dynamics during the formation of folding intermediates in these systems.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 我们小组之前在 APS 的研究表明，溶液中蛋白质的广角 X 射线散射 (WAXS) 对二级、三级和四级蛋白质结构的变化高度敏感。当应用于动态或灵活的系统（例如部分折叠的蛋白质或通常不适合 X 射线晶体学的结构状态集合）时，它特别强大。尽管有大量关于蛋白质从天然状态到熔球状态再到未折叠状态的转变的文献，但对于中间熔球状态中存在什么结构以及有多少结构仍然缺乏了解。该提案旨在利用 WAXS 对已充分表征的蛋白质的天然状态、熔球状态和变性状态进行分析，以阐明熔球状态的性质。 WAXS 数据将从脱辅基肌红蛋白等蛋白质中收集；乳清蛋白、泛素和乳球蛋白被诱导发生熔球形成和变性。使用我们在蛋白质呼吸研究中开发的分析方法，我们将表征每种蛋白质的熔球状态的动力学。这些结果将与芝加哥大学 T. Sosnick 进行的理论结构集合预测进行比较。这些研究之后将对所研究的蛋白质组内更稳定（就寿命而言）中间体进行时间分辨WAXS研究，以便跟踪这些系统中折叠中间体形成过程中结构和动力学的变化。