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.

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。 我们在APS的研究小组以前的研究表明，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，如部分折叠的蛋白质或结构状态的集合时，它是特别强大的，这些系统通常不适合X射线晶体学。虽然有相当多的关于蛋白质从天然到熔融球再到未折叠状态的转变的文献，但仍然存在对中间熔融球状态中存在什么和多少结构的理解相当缺乏。该提案旨在使用WAXS分析的天然，熔融球和变性状态的良好表征的蛋白质，以阐明熔融球状态的性质。WAXS数据将从蛋白质中收集，如脱辅基肌红蛋白、乳白蛋白、泛素和乳球蛋白，这些蛋白质已被诱导进行熔融球形成和变性。使用我们在蛋白质呼吸研究中开发的分析方法，我们将描述这些蛋白质中每一种的熔融球状态的动力学特征。这些结果将与T.芝加哥大学的索斯尼克。这些研究之后将进行时间分辨WAXS研究，研究蛋白质组中更稳定的（就寿命而言）中间体，以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。