A Novel SFM-based Method for Studying Single Molecule Dynamics

一种基于 SFM 的单分子动力学研究新方法

• 批准号: 7944647
• 负责人: Sean B. Andersson
• 金额: $ 12.5万
• 依托单位: BOSTON UNIVERSITY (CHARLES RIVER CAMPUS)
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-15 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7944647
• 关键词: Actins; Address; Algorithms; Arts; Atomic Force Microscopy; Binding; Biological; Biopolymers; Cells; DNA; DNA-Directed RNA Polymerase; Detection; Development; Dynein ATPase; Feedback; Foundations; Frequencies; Genetic Transcription; Goals; Head; Hereditary Disease; Image; In Vitro; Laws; Life; Liquid substance; Location; Measurement; Methods; Microscope; Microscopy; Microtubules; Modeling; Molecular Biology; Molecular Motors; Monitor; Motion; Motor; Myosin Type V; Optics; Pattern; Polymerase; Polymers; Positioning Attribute; Process; Proteins; Research; Resolution; Sampling; Scanning; Scanning Probe Microscopes; Scheme; Solutions; Speed; Structure; System; Techniques; Testing; Time; Tropomyosin; Update; Work; base; cantilever; design; improved; interest; millisecond; nanometer; nanoscale; novel; novel strategies; optical traps; particle; public health relevance; repair enzyme; research study; simulation; single molecule; theories; time use; tool

DESCRIPTION (provided by applicant): We propose to develop, analyze, implement and test a novel approach for tracking single particles in a scanning force microscope. The ability to study the dynamics of single molecules and of the interactions between molecules is a critical component for continued progress in molecular biology and for understanding and treating a variety of genetic diseases. Current techniques include using single particle tracking in optical microscopy and position tracking in optical traps. Particle tracking in optical microscopy is limited in its temporal resolution. Optical traps can provide superb spatial and temporal resolution; the construction of systems with such sensitivity, however, is extremely challenging. Due to the exquisite spatial resolution of scanning force microscopy as well as its ability to operate in liquid, it has become a standard tool for studying the structure of single molecules. The standard approach for studying dynamics is the use of time-lapse imaging. Each image can take seconds to minutes to acquire and thus the applicability of this approach is extremely limited. The exploratory research in this proposal is focused on developing feedback control algorithms for an atomic force microscope to directly track a single molecule such as a motor protein or a polymerase. We aim to (1) design and test algorithms for rapidly moving the tip along a biological polymer such as RNA, microtubules, and actin, without imaging, (2) combine these algorithms with detection and estimation schemes to track molecules moving on such structures, such as molecular motors or polymerases, and (3) apply the scheme to study the motion of tryopomyosin and of myosin V along actin. PUBLIC HEALTH RELEVANCE: In this project we propose to develop a novel method for studying the dynamics of single molecules moving on biopolymers. The method is a new control approach centered on the concept of particle tracking with a scanning force microscope. It takes advantage of the high spatial resolution of scanning force microscopy and the high temporal resolution inherent in the high resonant frequencies of the cantilevers.

描述（由申请人提供）：我们建议在扫描力显微镜中开发，分析，实施和测试一种新方法来跟踪单个颗粒。研究单分子和分子之间相互作用的动力学的能力是分子生物学以及理解和治疗多种遗传疾病的持续进展的关键成分。当前技术包括在光学显微镜和光学陷阱中的位置跟踪中使用单个粒子跟踪。光学显微镜中的粒子跟踪在其时间分辨率上受到限制。光学陷阱可以提供出色的空间和时间分辨率；但是，具有这种敏感性的系统的构建极具挑战性。由于扫描力显微镜的精美空间分辨率及其在液体中运行的能力，因此它已成为研究单分子结构的标准工具。研究动力学的标准方法是使用延时成像。每个图像可能需要秒到几分钟才能获取，因此该方法的适用性极为有限。该提案中的探索性研究重点是开发原子力显微镜的反馈控制算法，以直接跟踪单个分子（例如运动蛋白或聚合酶）。我们的目的是（1）设计和测试算法，以快速沿着生物聚合物（例如RNA，微管和肌动蛋白）迅速移动尖端，而无需成像，（2）将这些算法与检测和估计方案相结合，以跟踪分子在此类结构上进行的分子，例如分子运动或（3）运动均采用运动范围，并在此类结构上进行动作，并研究了该方案。 公共卫生相关性：在这个项目中，我们建议开发一种新的方法来研究在生物聚合物上移动的单分子的动力学。该方法是一种以扫描力显微镜的粒子跟踪概念为中心的新控制方法。它利用了扫描力显微镜的高空间分辨率和悬臂高谐振频率固有的高时间分辨率。