PROBING THE STRUCTURE OF THE FOLDING INTERMEDIATE OF CYTOCHROME C

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 了解蛋白质折叠和错误折叠的机制与许多人类疾病有关，并努力寻找治疗这些疾病的方法。为了更好地了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到自然状态，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，我们在两个方面集中了我们的努力。首先，为了提高数据收集效率，我们建造了一个自动采样器，以便在光束线上进行更高吞吐量的自动数据收集。随着样品处理能力的提高，我们已经对两种蛋白质(Chey和细胞色素c)的平衡变性进行了分析，这两种蛋白质解决了拓扑的作用和未折叠状态系综在指导折叠中的作用等基本问题。我们的团队还研究了锌金属离子和一种内在的二硫键HAS对铜锌超氧化物歧化酶(SOD)结构的影响，这是一种与Lou Gehrig病有关的蛋白质。我们的结果表明，在没有金属和二硫键交联的情况下，致病突变的SOD在生理温度下在结构上是展开的，并且可能更有可能聚集在一起。随着BioCAT的结构研究成为可能，我们也在研究致病突变对天然超氧化物歧化酶结构的影响。

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.