Protein structure, function and analysis in biofluids using ultrafast spectroscopy

背景资料：生物流体，如血清（去除红细胞的血液），提供了一个人健康的重要监测，因为它们含有蛋白质，糖，磷脂和核酸的复杂混合物，提供了代谢的分子指纹。测量生物流体分子组成的变化为监测健康和检测疾病提供了一种方便、信息丰富的方法。我们最近证明，超快2D-IR光谱是血清分析的有力工具。使用一系列超短红外激光脉冲，2D-IR可以抑制强烈的水分吸收，并拾取生物流体中蛋白质的信号，就像大海捞针一样（Hume et al Chemical Science 10，6448-6456（2019））。通过将红外光谱扩展到第二光谱维度（如2D-NMR），2D-IR还可以提供对蛋白质结构、结构的快速变化（结构动力学）和分子间相互作用极其敏感的2D“地图”。在这个项目中，我们将探索血清中蛋白质的结构、动力学和功能。通过了解如何2D-IR地图可以用来唯一地确定一个感兴趣的蛋白质的混合物，在确定如何生物流体的复杂分子环境影响其结构和动力学，这个博士将开发令人兴奋的新的见解和测量方法相关的化学，制药和生物医学领域。包括化学家、生物医学光谱学家和物理学家。该项目将测量一系列关键血清蛋白的2D-IR光谱，并开发测量其在真实世界样本中浓度的方法。利用2D-IR首次探测水中蛋白质动力学的能力，该项目将发展我们对身体生理环境如何影响蛋白质结构和动力学以及影响药物结合等过程的理解。

Background: Biofluids, such as blood serum (blood with the red blood cells removed), provide an important monitor of a person's health because they contain a complex mixture of proteins, sugars, phospholipids and nucleic acids that gives a molecular fingerprint of metabolism. Measuring changes in the molecular make-up of biofluids offers an easily accessible, informative way to monitor health and detect disease.We have recently shown that ultrafast 2D-IR spectroscopy is a powerful tool for blood serum analysis. Using a sequence of ultrashort infrared laser pulses, 2D-IR can suppress strong water absorptions and pick out signals due to proteins in biofluids like finding needles in a haystack (Hume et al Chemical Science 10, 6448-6456 (2019)). By spreading an infrared spectrum over a second spectral dimension (like 2D-NMR) 2D-IR also gives a 2D 'map' that is extremely sensitive to protein structure, rapid changes in structure (structural dynamics) and intermolecular interactions.In this project we will explore the structure, dynamics and function of proteins in blood serum. By understanding how the 2D-IR map can be used to uniquely identify a protein of interest in a mixture, before determining how the complex molecular environment of a biofluid affects its structure and dynamics, this PhD will develop exciting new insights and measurement methods relevant to the chemical, pharmaceutical and biomedical sectors.ObjectivesThis PhD project will be part of a multidisciplinary study developing 2D-IR for biomedical applications, including chemists, biomedical spectroscopists and physicists. The project will measure the 2D-IR spectra of a range of key blood serum proteins and develop methods to measure their concentrations in real-world samples. Using the ability of 2D-IR to probe protein dynamics in water for the first time, the project will develop our understanding of how the physiological environment of the body affects protein structure and dynamics and influences processes such as drug binding.