PROBING THE STRUCTURE OF THE FOLDING INTERMEDIATE OF CYTOCHROME C

细胞色素C折叠中间体结构的探讨

• 批准号: 8168637
• 负责人: Osman Bilsel
• 金额: $ 1.08万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168637
• 关键词: Address; Amyotrophic Lateral Sclerosis; Computer Retrieval of Information on Scientific Projects Database; Cuprozinc Superoxide Dismutase; Data Collection; Disease; Disulfides; Equilibrium; Funding; Grant; Institution; Ions; Metals; Physiological; Process; Property; Proteins; Research; Research Personnel; Resources; Role; Sampling; Source; Structure; Temperature; Therapeutic; United States National Institutes of Health; beamline; crosslink; cytochrome c; disease-causing mutation; human disease; improved; mutant; protein folding; protein structure

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. An understanding of the mechanism by which proteins fold and misfold is relevant to a number of human diseases and efforts to find therapeutics to treat them. To better understand the protein folding process and why proteins sometimes misfold, our efforts are aimed at understanding the fundamental principles guiding the acquisition of the native functional protein structure. Many proteins fold to the native state via intermediates, and small-angle scattering studies at BioCAT are instrumental in understanding their structural properties. In our studies this year at BioCAT, we have focused our efforts in two ways. First, to improve data collection efficiency, we have built an autosampler to allow higher-throughput automated data collection at the beamline. With the increased sample processing capability, we have performed an analysis of the equilibrium denaturation of two proteins (CheY and cytochrome c) that address fundamental questions on the role of topology and the role of the unfolded state ensemble in guiding folding. Our group has also studied the effect of Zn metal ions and an intrinsic disulfide cross-link has on the structure of Cu,Zn-superoxide dismutase (SOD), a protein implicated in Lou Gehrig's disease. Our results suggest that, in the absence of metals and the disulfide cross-link, disease causing mutants of SOD are structurally unfolded, and possibly more likely to aggregate, at physiological temperatures. With the structural studies made possible by BioCAT, we are also in the process of addressing the effect of disease causing mutations on the structure of the native state of SOD.

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。 了解蛋白质折叠和错误折叠的机制与许多人类疾病以及寻找治疗方法的努力有关。 为了更好地理解蛋白质折叠过程以及为什么蛋白质有时会错误折叠，我们的努力旨在了解指导获得天然功能蛋白质结构的基本原则。 许多蛋白质通过中间体折叠成天然状态，BioCAT的小角散射研究有助于了解它们的结构特性。 在我们今年在BioCAT的研究中，我们集中精力在两个方面。 首先，为了提高数据收集效率，我们构建了一个自动进样器，以允许在光束线处进行更高通量的自动数据收集。 随着样品处理能力的提高，我们对两种蛋白质（CheY和细胞色素c）的平衡变性进行了分析，这些蛋白质解决了拓扑结构的作用和未折叠状态集合在指导折叠中的作用的基本问题。 我们的小组还研究了锌金属离子和内在的二硫键交联对铜，锌-超氧化物歧化酶（SOD），一种与卢伽雷氏病有关的蛋白质的结构的影响。 我们的研究结果表明，在没有金属和二硫键交联，致病突变体的SOD结构展开，并可能更容易聚集，在生理温度。 随着BioCAT的结构研究成为可能，我们也在解决引起疾病的突变对SOD天然状态结构的影响。