A novel biochemical nanomotor

描述（由申请人提供）：纳米马达将化学或其他能源转化为运动，施加可以移动外部货物的力。它们是纳米技术的重要组成部分。该项目的总体目标是开发一种沿着线性聚合物移动的生化纳米马达。这种器件将用作动力纳米器件内的模块。它将被控制和指挥，以便沿着指定的轨道将货物运送到特定的地址。为此，我们将使用依赖atp的蛋白质ClpX，它接合、展开和移动蛋白质。在其原生细菌环境中，ClpX的功能是将蛋白质底物喂入蛋白酶。我们将调整它沿着DNA移动，从而使用ClpX作为atp驱动的绞车，DNA作为电缆。为了实现这些目标，我们将进行以下实验。具体目标1：构建嵌合物，包括：(1)与ClpX结合并启动易位的蛋白质或肽，连接到(2)ClpX随后继续移动的DNA。具体目标2：开发和应用分析来评估ClpX沿着DNA的动力易位。具体目标3：使用这些测定，修改DNA的性质，以优化易位速度和功率。利用大肠杆菌ClpX和来自不同细菌物种的ClpX诱变来改善电机的预期性能。

DESCRIPTION (provided by applicant): Nanomotors convert chemical or other energy sources into motion, exerting forces that can move an external cargo. They are an important part of nanotechnology. The overall goal of this project is to develop a biochemical nanomotor that moves along a linear polymer. Such a device will be used as a module within a powered nanodevice. It will be controlled and directed so as to deliver cargo along specified tracks to specific addresses. To that end we will use the ATP-dependent protein ClpX, which engages, unfolds and moves proteins. In its native bacterial context ClpX functions to feed protein substrates into a protease. We will adapt it to move instead along DNA, thereby using ClpX as an ATP-powered winch with DNA as a cable. In furtherance of these goals we will perform the following experiments. Specific Aim 1: Construct chimeric objects consisting of (1) a protein or peptide that engages ClpX and initiates translocation, linked to (2) a DNA along which ClpX subsequently continues to move. Specific Aim 2: Develop and apply assays to assess the powered translocation of ClpX along DNA. Specific Aim 3: Using these assays, modify the properties of DNA to optimize translocation velocity and power. Use mutagenesis of E. coli ClpX and ClpX sourced from diverse bacterial species to improve the desired properties of the motor. PUBLIC HEALTH RELEVANCE: oject Narrative Nanomotors convert chemical or other energy sources into motion, exerting forces that can move an external cargo. They are an important part of nanotechnology. We will adapt the bacterial protein ClpX, which engages, unfolds and moves proteins, developing it to function as a novel biochemical nanomotor that moves along DNA.