In-cell Structural Biology using Electron Paramagnetic Resonance Spectroscopy

使用电子顺磁共振波谱进行细胞内结构生物学

• 批准号: 1996168
• 金额: --
• 依托单位: University of St Andrews
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1996168
• 关键词: cell; Structural; Biology; using; Electron

The aim of this research is to understand the interactions that occur within the proteome and as a consequence, the interactions required for life. To achieve this, we first need to understand the structures and modes of action of the proteins involved. This project uses electron paramagnetic resonance (EPR) spectroscopy to determine how proteins fit together and in turn how this structure enables their function. EPR allows us to measure the energies of electron spins, and so determine the interactions they have with each other. The interactions change dependent on the distance between spins, and so we can use EPR to infer spin separation. By attaching spins to proteins at specific positions we can determine how proteins physically interact with one another. EPR is best suited to measuring nanometre distances, and so is a tool that is complementary to the more familiar methods of X-ray crystallography, electron microscopy and NMR. To-date most structural studies of proteins using EPR have looked at the proteins outside the cellular environment. This gives us a good indication of the function, but doesn't take into account interactions and changes in protein behaviour as part of a larger, regulated system. There is great scope to use EPR for measurements inside cells and thus obtain structural information in a native environment. The motivation for this research into the biomolecular action of proteins is to determine with quantitative evidence how proteins perform their roles in the cell and how changes to this mode of action can give rise to disease. We will be working with a variety of different cell lines, proteins and spin labels to test the applicability of our methodology as well as the extent to which it can be applied.

这项研究的目的是了解蛋白质组内发生的相互作用，从而了解生命所需的相互作用。要做到这一点，我们首先需要了解所涉及的蛋白质的结构和作用模式。该项目使用电子顺磁共振(EPR)光谱学来确定蛋白质是如何结合在一起的，进而确定这种结构如何使它们发挥功能。EPR使我们能够测量电子自旋的能量，从而确定它们之间的相互作用。相互作用的变化取决于自旋之间的距离，因此我们可以使用EPR来推断自旋分离。通过在特定位置将自旋连接到蛋白质上，我们可以确定蛋白质如何在物理上相互作用。EPR最适合测量纳米距离，因此是一种补充更常见的X射线结晶学、电子显微镜和核磁共振方法的工具。到目前为止，使用EPR对蛋白质的结构研究大多着眼于细胞环境之外的蛋白质。这给了我们一个很好的功能指示，但没有考虑到作为更大的、受调节的系统的一部分的相互作用和蛋白质行为的变化。EPR在细胞内的测量中有很大的应用范围，因此可以在自然环境中获得结构信息。这项研究蛋白质的生物分子作用的动机是用定量的证据确定蛋白质如何在细胞中发挥其作用，以及这种作用模式的变化如何导致疾病。我们将使用各种不同的细胞系、蛋白质和自旋标记来测试我们方法的适用性以及它可以应用的程度。