In Vitro Dynamics of Kinesin 1 and Myosin VI

驱动蛋白 1 和肌球蛋白 VI 的体外动力学

• 批准号: 8037008
• 负责人: PAUL R SELVIN
• 金额: $ 34.58万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-06-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8037008
• 关键词: ATP phosphohydrolase; Address; Affect; Alzheimer' s Disease; Amino Acids; Antineoplastic Agents; Binding; CENP-E protein; Calmodulin; Cells; Central Nervous System Diseases; Chimera organism; Color; Cysteine; Data; Defect; Development; Dimensions; Dimerization; Family member; Fluorescence; Grant; Hand; Head; Head and neck structure; Health; Heart Diseases; Image; In Vitro; Kinesin; Kinetics; Length; Leucine Zippers; Light; Link; Maintenance; Malignant neoplasm of brain; Measures; Microtubules; Molecular Motors; Motion; Motor; Motor Activity; Movement; Myosin ATPase; Neck; Nucleotides; Occupations; Organelles; Paper; Pharmaceutical Preparations; Phase; Physiological; Play; Positioning Attribute; Probability; Protein Region; Proteins; Regulation; Resolution; Rice; Role; Running; SWI1; Science; Seminal; Signal Transduction; Structure; Tail; Techniques; Testing; Time; Torsion; Walking; Work; arm; cancer type; cell motility; crosslink; deafness; falls; fluorescence imaging; fluorophore; graduate student; millisecond; mutant; myosin VI; nanometer; optical traps; single molecule; single-molecule FRET; small molecule; tool

DESCRIPTION (provided by applicant): Molecular motors--kinesin and myosins--play a crucial role in the maintenance and development of the organization, motility, and signaling of healthy cells. Central nervous-system disorders, such as Alzheimer's disease and certain types of cancer, all arise from molecular motors gone awry. There are, however, several fundamental questions about how kinesin-1 moves. (Kinesin-1 is the "standard-bearer" of kinesins.) Also largely unknown is how Myosin VI moves. To answer these questions we will address the motors at the in vitro, single molecule level. We will apply a number of single molecule tools, including some that have been developed in the first 4 years of this proposal. We will use FIONA (Fluorescence Imaging with One Nanometer Accuracy), DOPI (Defocused Orientation and Position Imaging), and SHREC (Single Molecule High REsolution Colocalization). The theme of the kinesin part is: how is motility affected by each of kinesin's parts, including the head, the coiled-coil, and the tail? The theme of the myosin VI part is: how does such a small motor take such a large step? We will present unpublished data which suggests that kinesin is bound by both heads during a run, making the probability of falling off very low. We will also suggest that the large non-helical region in kinesin-1's coiled-coiled region allows this motor to walk in an "asymmetric fashion," allowing the cargo to point forward. Finally, we will suggest that full-length kinesin takes "pauses" due to the tail-region folding over and interacting with the head region and possibly the microtubule. With regards to myosin VI, we have an enormous amount of preliminary data. It will suggest that the head undergoes a 180¿ swing during the powerstroke. Furthermore, we suggest that a 3-helix motif in the lever arm uncoils and creates an unprecedented 24 nm extension, which allows the motor to take a 36 nm step. PUBLIC HEALTH RELEVANCE Molecular motors have the job of moving and organizing organelles within a cell. Problems within the motors cause brain cancer, Alzheimer's disease, etc. We seek to understand the basic workings of kinesins and myosin VI, two important motors.

描述（由应用提供）：分子电动机 - 运动蛋白和肌球蛋白 - 在组织的维持和发展，健康细胞的信号和信号传导中扮演至关重要的作用。中枢神经系统疾病（例如阿尔茨海默氏病和某些类型的癌症）都源于分子电机出现了。但是，关于运动蛋白-1的移动有几个基本问​​题。 （驱动蛋白-1是驱动蛋白的“标准杆”。）也未知的是肌球蛋白VI的移动方式。为了回答这些问题，我们将在体外单分子水平上解决电动机。我们将应用许多单分子工具，包括在该提案的前4年中开发的一些工具。我们将使用fiona（具有一个纳米精度的荧光成像），dopi（散落的方向和位置成像）和SHREC（单分子高分辨率）共定位）。动力素部分的主题是：运动能力如何受到运动蛋白的各个部分的影响，包括头部，盘绕螺旋和尾巴？肌球蛋白VI部分的主题是：如此小的电动机如何迈出如此大的一步？我们将提供未发表的数据，这表明动力蛋白在运行期间都受两个头部的束缚，从而使跌落很低的概率非常低。我们还将建议，在动机1的盘绕区域中的大型非螺旋区域使该电动机以“不对称方式”行驶，从而使货物指向前进。最后，我们建议全长驱动蛋白由于尾区折叠并与头部区域相互作用以及可能的微管而进行“停顿”。关于肌球蛋白VI，我们有大量的初步数据。这表明头部在力量中经历了180次摇摆。此外，我们建议杠杆臂中的3螺旋图案脱落并创建前所未有的24 nm延伸，这使电动机可以采取36 nm的步骤。公共卫生相关性分子电动机具有在细胞内移动和细胞器的工作。电动机中的问题会导致脑癌，阿尔茨海默氏病等。我们试图了解两个重要电机的驱动蛋白和肌球蛋白VI的基本起作用。