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）：我们建议开发、分析、实施和测试一种在扫描力显微镜中跟踪单个粒子的新方法。研究单分子动力学和分子间相互作用的能力是分子生物学持续进步以及理解和治疗各种遗传疾病的关键组成部分。当前的技术包括在光学显微镜中使用单粒子跟踪和在光阱中使用位置跟踪。光学显微镜中的粒子追踪因其时间分辨率而受到限制。光陷阱可以提供卓越的空间和时间分辨率；然而，构建具有如此敏感性的系统极具挑战性。由于扫描力显微镜精致的空间分辨率以及在液体中操作的能力，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。每张图像可能需要几秒到几分钟的时间来获取，因此这种方法的适用性极其有限。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。我们的目标是 (1) 设计和测试算法，用于在不成像的情况下沿着生物聚合物（如 RNA、微管和肌动蛋白）快速移动尖端，(2) 将这些算法与检测和估计方案结合起来，以跟踪在此类结构上移动的分子，例如分子马达或聚合酶，以及（3）应用该方案来研究试肌球蛋白和肌球蛋白V沿着肌动蛋白的运动。 公共健康相关性：在这个项目中，我们建议开发一种新方法来研究单分子在生物聚合物上移动的动力学。该方法是一种以扫描力显微镜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共振频率固有的高时间分辨率。