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Structural basis of motility by dimeric kinesin and myosin motor proteins

二聚驱动蛋白和肌球蛋白运动蛋白运动的结构基础

基本信息

批准号：
10533263
负责人：
CHARLES VAUGHN SINDELAR
金额：
$ 34.39万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-05-01 至 2023-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10533263
关键词：
3-Dimensional Actins Active Sites Address Automobile Driving Binding Biochemical Cardiomyopathies Cell division Cells Chemical Structure Chemicals Classification Communication Complex Consumption Couples Cryoelectron Microscopy Cytokinesis Cytoskeletal Filaments Cytoskeleton DNA Structure Data Defect Development Dimerization Disease Electron Microscope Filament Freezing Funding Generations Goals Head Hydrolysis Image Individual Investigation Kinesin Life Malignant Neoplasms Maps Mechanics Methods Microfilaments Microtubules Mitosis Molecular Molecular Conformation Molecular Machines Molecular Motors Motor Movement Myosin ATPase Myosin Type V Nature Neurotransmitters Nonmuscle Myosin Type IIA Nucleotides Pharmacologic Substance Power stroke Preparation Process Production Productivity Proteins Research Resolution Role Sampling Shapes Spastic Paraplegia Structure Synapses System Time Visualization Walking Work analog arm cancer therapy cell motility design dimer experimental study foot image processing improved inhibitor inorganic phosphate insight molecular rearrangement nanometer resolution nerve supply three dimensional structure three-dimensional visualization trait

项目摘要

Project Summary Kinesin and myosin are so-called `motor proteins' that can use two `feet' to walk along microtubule and actin filaments (respectively) that make up the cytoskeleton. These motors support many vital functions with the cell, including pulling DNA structures apart during cell division and resupplying nerve junctions (the synapse) with neurotransmitters (such as seratonin). The precise mechanisms by which these elaborate molecular machines, which are composed of tens of thousands of exquisitely arranged atoms, are able to `walk' are complex and incompletely understood. To see how they work, it is necessary to visualize these complex structures in three dimensions at sufficient levels of detail to resolve individual atoms– and to follow molecular rearrangements that happen while the motors step forward. This goal, however, has long remained out of reach due to the extreme technical challenges involved. We have addressed this problem by developing new methods to analyze images of frozen motor-filament assemblies collected by latest-generation electron microscopes. This approach, known as cryo-electron microscopy, allows us to directly visualize the three dimensional shape of individual molecular motor proteins attached to their partner filaments. During the previous funding period, we solved 3D structures of truncated single `feet' (one motor domain) of kinesin and myosin motors attached to their partner filaments, showing in atomic detail how these structures changed when molecules of ATP fuel were bound and consumed. We also captured a 3D structure of an intact pair of kinesin molecules (dimer) caught in mid- step on a microtubule. This allowed us to visualize, for the first time, a way in which the two `feet' of kinesin can pull on each other in a way to stay coordinated while walking. In our ongoing research we are improving our methods to capture more intermediates in the stepping process of kinesin, in order to gain a complete more understanding of how it walks. We are improving our analysis methods to better resolve precise chemical details within these structures. Finally, we are extending our approach to understand how a pair of myosin molecules can walk along the actin filament. Results of our studies are expected to aid the development of a new generation of pharmaceutical agents for treating cancer and a wide variety of other diseases.

项目摘要肌动蛋白和肌球蛋白是一种所谓的“马达蛋白”，它们可以用两只“脚”走路。构成细胞骨架的微管和肌动蛋白细丝。这些马达支持细胞的许多重要功能，包括在细胞过程中拆分DNA结构神经接头(突触)与神经递质(如血清素)。这些精密的分子机器的精确机制，是由数以万计排列精巧的原子组成，能够‘行走’的都是复杂的也不能完全理解。为了了解它们是如何工作的，有必要将这些复杂的东西可视化以足够详细的水平分解单个原子的三维结构--以及跟随马达前进时发生的分子重排。这个目标，然而，由于涉及的极端技术挑战，长期以来一直遥不可及。我们已经通过开发新的方法来分析冻结的图像来解决这个问题用最新一代电子显微镜收集的电机细丝组件。这种方法，被称为冷冻电子显微镜，使我们能够直接看到三维形状单个分子马达蛋白附着在它们的伴侣细丝上。在之前的资助期间，我们解决了截断的单足(一个马达)的3D结构结构域)连接到它们的配对细丝上的动蛋白和肌球蛋白马达，以原子表示详细说明当ATP燃料分子被结合和消耗时，这些结构是如何变化的。我们还捕捉到了一对完整的动蛋白分子(二聚体)的3D结构，这些分子被捕捉到踩在微管上。这让我们第一次看到，这两种方式动蛋白的“脚”可以相互牵拉，以保持行走时的协调性。在我们正在进行的研究我们正在改进我们的方法，以在分步过程中捕获更多的中间体为了更全面地了解它是如何行走的。我们正在改进我们的分析方法可以更好地解析这些结构中的精确化学细节。最后，我们正在扩展我们的方法，以了解一对肌球蛋白分子是如何行走的肌动蛋白细丝。我们的研究结果有望帮助开发一种新的用于治疗癌症和各种其他疾病的新一代药物。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A vertebrate myosin-I structure reveals unique insights into myosin mechanochemical tuning.

脊椎动物肌球蛋白-I 结构揭示了对肌球蛋白机械化学调节的独特见解。

DOI：
10.1073/pnas.1321022111
发表时间：
2014
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Shuman,Henry;Greenberg,MichaelJ;Zwolak,Adam;Lin,Tianming;Sindelar,CharlesV;Dominguez,Roberto;Ostap,EMichael
通讯作者：
Ostap,EMichael

Structural basis of fast- and slow-severing actin-cofilactin boundaries.

DOI：
10.1016/j.jbc.2021.100337
发表时间：
2021-01
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Hocky GM;Sindelar CV;Cao W;Voth GA;De La Cruz EM
通讯作者：
De La Cruz EM

High-resolution structures of kinesin on microtubules provide a basis for nucleotide-gated force-generation.

DOI：
10.7554/elife.04686
发表时间：
2014-11-21
期刊：
eLife
影响因子：
7.7
作者：
Shang Z;Zhou K;Xu C;Csencsits R;Cochran JC;Sindelar CV
通讯作者：
Sindelar CV

Twist response of actin filaments.

肌动蛋白丝的扭曲反应。

DOI：
10.1073/pnas.2208536120
发表时间：
2023-01-24
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
通讯作者：

Calcium sensitive ring-like oligomers formed by synaptotagmin.

由突触结合蛋白形成的钙敏感环状寡聚物。

DOI：
10.1073/pnas.1415849111
发表时间：
2014
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Wang,Jing;Bello,Oscar;Auclair,SarahM;Wang,Jing;Coleman,Jeff;Pincet,Frederic;Krishnakumar,ShyamS;Sindelar,CharlesV;Rothman,JamesE
通讯作者：
Rothman,JamesE