Ultra-stable, high-bandwidth measurement platform for high-precision studies of rapid conformational dynamics in single biomolecules

超稳定、高带宽测量平台，用于单个生物分子快速构象动力学的高精度研究

• 批准号: RTI-2016-00172
• 负责人: Woodside, Michael
• 金额: $ 7.28万
• 依托单位: University of Alberta
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=591366
• 关键词: Ultra; stable; bandwidth; measurement; platform

Biological molecules like proteins, DNA, and RNA form complex structures that are required for them to function correctly. Understanding how these structures form (known as ‘folding’) is important for understanding both proper function and dysfunction (which may lead to disease). One of the most sensitive ways to study folding is to use laser tweezers to grab onto single molecules and unravel their structure by pulling them apart. This proposal will build an ultra-stable system for measuring the folding of single molecules, one that can observe very rapid (~1 μs) events with atomic-scale resolution over long periods of time (tens of minutes), providing the most precise measurements to date of real-time structural changes in these molecules. We will use it to study fundamental questions such as how folding works as a physical process and what determines the specific behaviour observed. More specifically, we will study the very shortest-lived events during folding, the elusive ‘transition states’ that act as bottlenecks in the process and therefore dominate the dynamics. The path taken through the transition states contains all the critical information about the mechanism of folding, but it is very hard to detect. We will measure these ‘transition paths’ not only to learn about the basic physical principles of folding, but also to understand how folding goes wrong: such misfolding is at the root of many incurable diseases, like Alzheimer’s and ALS, but the mechanisms that drive it at the molecular level remain poorly understood. Furthermore, we will use the new instrument to study still-unsolved biological processes like how viruses can alter the way the genetic code is read, which pose important scientific riddles but also have potential applications in medicine. Studies like these, making use of the incredible resolution and sensitivity of single-molecule probes, are one of the hottest frontiers in folding studies. We have right now a remarkable opportunity to make significant and lasting advances in a decades-old problem with relevance to biology, physics, chemistry, and medicine. Without this instrumentation, we will not be able to pursue such exciting science.

蛋白质，DNA和RNA之类的生物分子形成了它们正确起作用所需的复杂结构。了解这些结构的形成（称为“折叠”）对于理解适当的功能和功能障碍（可能导致疾病）很重要。研究折叠的最敏感方法之一是使用激光镊子抓住单分子并通过将它们拉开来揭开其结构。该建议将建立一个超稳的系统，用于测量单分子的折叠，该系统可以在长时间（数十分钟）内观察到具有原子尺度分辨率的非常快速（〜1μs）的事件，从而提供了这些分子中实时结构变化以后的最精确测量。我们将使用它来研究基本问题，例如折叠方式如何作为物理过程以及确定观察到的特定行为的原因。更具体地说，我们将研究折叠期间最短的事件，即在此过程中起瓶颈的难以捉摸的“过渡状态”，因此主导了动态。通过过渡状态采取的路径包含有关折叠机制的所有关键信息，但很难检测到。我们将衡量这些“过渡路径”，不仅是为了了解折叠的基本物理原理，而且还要了解折叠是如何出现的：这种错误折叠是许多无法治愈的疾病的根源，例如阿尔茨海默氏症和ALS，而且在分子水平上驱动它的机制仍然不足。此外，我们将使用新工具来研究仍未解决的生物学过程，例如病毒如何改变遗传密码的阅读方式，从而构成了重要的科学谜语，但在医学中也有潜在的应用。这样的研究是利用单分子问题的令人难以置信的分辨率和敏感性，是折叠研究中最热门的边界之一。现在，我们有一个非凡的机会，可以在与生物学，物理学，化学和医学相关的数十年问题中取得重大而持久的进步。没有这种仪器，我们将无法追求这种令人兴奋的科学。