An Optical Single Molecule Scanner of Protein Motion

尽管X射线晶体学和电子显微镜技术取得了巨大的进步，但我们还没有一种方法来可视化运动中的功能蛋白质。该奖学金将引领所需的突破，并开发出第一种光学仪器，以实时和单分子水平可视化蛋白质。我们建议开发一种仪器，以特定和灵敏的方式探测单个蛋白质，同时尽可能少地干扰它们。我们的愿景是创造一种“分子扫描仪”，可以扫描任意蛋白质及其动力学，这是一种超越当前最先进技术的技术。实现这种传感器将导致对生命机制如何运作的新的基本理解。微型光学传感器将使我们能够以全新的方式分析蛋白质。我们将能够在便携式硬币大小的实验室中通过光致振动运动来检测蛋白质。我设想的进展将导致一种全新的方法，用于分析和诊断蛋白质错误折叠疾病（蛋白质病），如朊病毒病，阿尔茨海默病，帕金森病，淀粉样变性和其他各种疾病。我们的传感器平台将能够通过提供实验室测试床来观察纳米机器的真实的运动，从而为人工分子机器的发展做出贡献。我们将利用光等离子体传感器实现这一仪器。光等离子体传感器通过光的反射驱动循环来增强检测信号。他们将光集中在纳米级，探测单个蛋白质。我们的目标是扫描单个蛋白质的纳米级光场，以提供有关蛋白质结构及其动力学的信息，在纳秒的时间尺度和单键和原子的空间尺度上解析蛋白质运动和振动。这项研究开发的光学技术将在蛋白质分析中开辟全新的领域。它将测量和可视化蛋白质结构及其原位，溶液和表面的动力学。它将完成蛋白质组学的“圣杯”之一。此外，这种技术可以集成在芯片上，允许从微量样品中识别错误折叠的蛋白质，样品制备最少。因此，它将为制药和化学分析行业创造新的分析方法，生物标志物和标准。许多行业将受益于该奖学金的进步，包括分析传感仪器，一个484亿美元的国际市场。医学界迫切需要这种分析工具来快速检测和识别内在无序的蛋白质，这些蛋白质会导致使人衰弱的蛋白质病，如帕金森病和阿尔茨海默病，影响全球超过4700万人，每年的医疗费用约为6040亿美元（WHO 2017）。

Despite dramatic advances in x-ray crystallography and electron microscopy, we do not have a way to visualise functional proteins in motion.This fellowship will lead the required breakthroughs and develop the first optical instrument to visualise proteins in real-time and at the level of single molecules. We propose to develop an instrument to probe single proteins in a specific and sensitive manner, while disturbing them as little as possible. The vision is to create a 'molecular scanner' that can characterise an arbitrary protein and its dynamics, a technology that is beyond the current state-of-the-art. Realising this sensor will lead to a new fundamental understanding of how the machinery of life functions. The micro-optical sensor will allow us to analyse proteins in entirely new ways. We will be able to detect proteins specifically, from optically-induced vibrational motions, on portable coin-sized laboratories. The advances I envisage will result in a completely new approach for the analysis and diagnosis of protein-misfolding diseases (proteinopathies) such as prion diseases, Alzheimer's disease, Parkinson's disease, amyloidosis, and a wide range of other disorders. Our sensor platform will be able to contribute to the development of artificial molecular machinery by providing laboratory test beds that observe the motions of nano-machines in real time.We will realise this instrument with optoplasmonic sensors. Optoplasmonic sensors enhance detection signals by reflection-driven circulation of the light. They concentrate the light at the nanoscale where they probe single proteins. We aim to scan the nanoscale light field across a single protein to provide information on the protein structure and its dynamics, resolving protein motions and vibrations at a temporal scale of nanoseconds and at a spatial scale of single bonds and atoms. The optical technique developed in this fellowship will instigate entirely new domains in protein analysis. It will measure and visualise protein structure and its dynamics in-situ, in solution and at surfaces. It will accomplish one of the "holy-grails" of proteomics. Also, this technique can be integrated on a chip, allowing the identification of misfolded proteins from a trace amount of sample, with minimal sample preparation. Thereby it will create new analysis methods, biomarkers and standards for the pharmaceutical and chemical analysis industries.A multitude of industries will be benefitted by the advances of this fellowship, including analytical sensing instrumentation, a $48.4 billion international market. The medical community desperately needs this analysis tool to rapidly detect and characterise intrinsically disordered proteins which cause the debilitating proteinopathies such as Parkinson's and Alzheimer's disease affecting more than 47 million worldwide, at an annual healthcare cost of ~$604 billion (WHO 2017).