Development of Biomolecular Analysis methods for 2D-IR Spectroscopy

该博士课程的目标是继续发展二维红外光谱工具和数据分析方法，使用基本的动态数据来告知分析技术的建设。实验方法将利用stfc中央激光设施ULTRA激光系统。我们将在先前的合作工作基础上，寻求对小分子与蛋白质和DNA结合的科学理解，同时提高英国国家设施的能力。该项目分为3个部分：第一部分：生物分子复合物的高通量2D-IR光谱，该部分将开发高通量2D-IR方法用于观察小分子与蛋白质的结合。这将把最近dna结合系统的研究扩展到光谱上更复杂和微妙的蛋白质-药物结合问题。特别是，我们将利用2D-IR来报告药物结合时蛋白质动力学和分子内振动耦合的变化，将光谱可观察到的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实时测量。该链将开发改进的t -跳红外和t -跳二维红外光谱为基础的策略，允许观察生物分子的动态非平衡响应在温度的步骤，但扩展他们允许联合加热/冷却实验。这些将用于观察溶液中动态生物分子结构变化的例子，无论是在初始扰动期间还是在松弛时，都是如此，为配体定位结合位点的过程提供了新的途径，从而直接观察诸如诱导契合或构象选择等机制。我们将从双链DNA或RNA的融化/重结合作为测试系统开始，包括配体结合事件，但在可以探测蛋白质二级结构变化和配体结合的实验方面取得进展，可能会从1链中提取关键系统。这条线将应用CLF的尖端激光设备来推进我们对溶液中生物分子动态行为的理解。链3：这个雄心勃勃的最后阶段将结合链1和链2的结果，通过开发蛋白质- dna结合的测量，最初是在平衡状态，但可能在动态条件下。

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.