Development of Biomolecular Analysis methods for 2D-IR Spectroscopy

二维红外光谱生物分子分析方法的开发

• 批准号: 2602917
• 金额: --
• 依托单位: University of York
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2602917
• 关键词: Development; Biomolecular; Analysis; methods; 2D

The aim of this PhD programme is to continue the development of 2-dimensional infrared spectroscopy tools and data analysis methods, using fundamental dynamic data to inform construction of analytical technologies. Experimental methods will utilise the STFC-Central Laser Facility ULTRA laser systems. We will seek to build on previous collaborative work towards scientific understanding of binding small molecules to proteins and DNA while advancing the capability of UK national facilities. The project is in 3 strands: Strand 1: High throughput 2D-IR spectroscopy of biomolecular complexesThis strand will develop the use of high throughput 2D-IR methods for observing small molecule binding to proteins. This will extend recent work on DNA-binding systems to the spectroscopically more complex and subtle problem of protein-drug binding. In particular, we will target the use of 2D-IR to report on changes in protein dynamics and intramolecular vibrational coupling upon drug binding, linking spectral observables to biological function or physical phenomena such as binding constants. Strand 2: Real time measurements of biomolecular interactions.This strand will develop modified T-jump IR and T-jump 2D-IR spectroscopy-based strategies that allow observation of dynamic non-equilibrium responses of biomolecules to steps in temperature, but extend them to allow combined heating/cooling experiments. These will be used to observe examples of dynamic biomolecular structure change in solution both during the initial perturbation but also upon relaxation, giving new access to the process of location of the binding site by the ligand and so direct observation of mechanisms such as induced fit or conformational selection. We will begin with melting/rebinding of double stranded DNA or RNA as test systems including ligand binding events, but progress towards experiments that can probe protein secondary structure change and ligand binding, potentially drawing on key systems from Strand 1.This strand will apply the cutting edge laser equipment of the CLF to advance our understanding of dynamic behaviour of biological molecules in solution. Strand 3: This ambitious final stage will combine the results of Strand 1 and 2 by developing measurements of protein-DNA binding, initially at equilibrium but potentially under dynamic conditions.

该博士课程的目的是继续开发二维红外光谱工具和数据分析方法，使用基本动态数据为分析技术的构建提供信息。实验方法将利用STFC中央激光设施ULTRA激光系统。我们将寻求建立在以前的合作工作对结合小分子蛋白质和DNA的科学理解，同时推进英国国家设施的能力。该项目分为三个部分：第一部分：生物分子复合物的高通量2D-IR光谱学这一部分将开发高通量2D-IR方法，用于观察小分子与蛋白质的结合。这将扩展最近的工作DNA结合系统的光谱更复杂和微妙的蛋白质药物结合的问题。特别是，我们将有针对性地使用2D-IR来报告药物结合后蛋白质动力学和分子内振动耦合的变化，将光谱观测值与生物功能或物理现象（如结合常数）联系起来。链2：生物分子相互作用的真实的时间测量。这条链将开发改进的T-jump IR和T-jump 2D-IR光谱为基础的策略，允许观察生物分子对温度步骤的动态非平衡响应，但将其扩展到允许组合加热/冷却实验。这些将被用来观察在溶液中的动态生物分子结构变化的例子，无论是在初始扰动，但也在放松时，给新的访问的过程中的结合位点的位置的配体和直接观察的机制，如诱导适合或构象选择。我们将开始与熔化/重新结合的双链DNA或RNA作为测试系统，包括配体结合事件，但进展的实验，可以探测蛋白质二级结构的变化和配体结合，潜在的借鉴关键系统从链1。这条链将应用CLF的尖端激光设备，以提高我们的理解的动态行为的生物分子在溶液中。链3：这个雄心勃勃的最后阶段将联合收割机链1和2的结果，通过开发蛋白质-DNA结合的测量，最初在平衡状态下，但可能在动态条件下。