Structure and function of nonmuscle myosins

非肌肉肌球蛋白的结构和功能

• 批准号: 10649564
• 负责人: KRISHNA CHINTHALAPUDI
• 金额: $ 36.54万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10649564
• 关键词: ATP phosphohydrolase; Acceleration; Actins; Address; Adopted; Architecture; Binding; Biochemical; Biological; Biological Assay; Biology; Cell Adhesion; Cell division; Cell physiology; Cells; Cellular biology; Computer software; Cryoelectron Microscopy; Cues; Cytoskeleton; Data; Defect; Development; Disease; Electron Microscopy; Engineering; Enzyme Activation; Enzymes; Equilibrium; Eukaryotic Cell; Filament; Fluorescence Microscopy; Foundations; Future; Generations; Goals; Health; Heart Diseases; Hematological Disease; Heterogeneity; Human; In Vitro; Kinetics; Knowledge; Length; Malignant Neoplasms; Measurement; Microfilaments; Modeling; Molecular; Molecular Conformation; Motor; Motor Activity; Mutation; Myosin ATPase; Myosin S-2; Nephritis; Nucleotides; Physiological Processes; Play; Process; Property; Protein Dephosphorylation; Protein Engineering; Proteins; Regulation; Research; Resolution; Role; Structure; Techniques; Testing; Textbooks; Therapeutic; Tissues; Work; X-Ray Crystallography; cell fixing; cell motility; computerized data processing; deafness; design; enzyme activity; flexibility; high resolution imaging; human disease; innovation; mechanotransduction; migration; mutant; nervous system disorder; non-muscle myosin; novel; paralogous gene; programs; protein complex; sensor; tool

Project Summary A hallmark of eukaryotic cells is their ability to migrate, divide, adhere and respond to environmental cues. Nonmuscle myosin-2 (NM2) motors play an essential role in many aspects of these fundamental cellular processes by forming short bipolar filaments that interact with actin filaments. NM2 motors are binary switches that alter between inactive and active states depending on the cellular context. The precise control of NM2 motor activity is critical for its cellular function as master regulator of the actin cytoskeleton. Aberrant regulation due to mutations in NM2 paralogs contribute to a whole host of diseases including blood and neurological disorders, heart diseases, deafness, nephritis, and cancers. NM2-specific therapies are thus needed, yet the lack of basic knowledge about the structure and regulation of NM2 paralogs is a bottleneck to their development. We aim to develop a detailed structural and mechanistic understanding of how force generation by NM2 motors drive various cellular functions. Using innovative and interdisciplinary techniques including the state-of-the-art cryo- electron microscopy, X-ray crystallography, steady-state kinetics, in vitro motility assays and high-resolution fluorescence microscopy, we will systematically dissect the mechanisms of activation and regulation of NM2. To achieve this, in Aim 1, we will determine the major structural states in the ATPase cycle of NM2 motors to explain enzyme function. In Aim 2, we will determine a high-resolution cryo-EM structure of the inactive state of full- length NM2 to explain its molecular architecture. In Aim 3, we will study the consequences of abolishing the ability to form an inactive state on the dynamics of NM2 filaments in cells. Collectively, our studies will provide a deeper understanding of the structure, function and regulation of NM2. Importantly, this knowledge will advance our understanding of emergent NM2 functions in cells and thus, lay the foundation for future development of NM2-specific therapeutics.

项目概要 真核细胞的一个标志是它们具有迁移、分裂、粘附和对环境信号做出反应的能力。 非肌肉肌球蛋白 2 (NM2) 马达在这些基本细胞的许多方面发挥着重要作用 通过形成与肌动蛋白丝相互作用的短双极丝来进行过程。 NM2电机是二元开关 根据细胞环境在非活动和活动状态之间变化。 NM2电机的精准控制 活性对其作为肌动蛋白细胞骨架的主要调节因子的细胞功能至关重要。由于监管异常 NM2旁系同源物的突变会导致一系列疾病，包括血液和神经系统疾病， 心脏病、耳聋、肾炎和癌症。因此需要 NM2 特异性疗法，但缺乏基础疗法 对 NM2 旁系同源物的结构和调控的了解是其发展的瓶颈。我们的目标是 对 NM2 电机如何产生驱动力有详细的结构和机械理解 各种细胞功能。使用创新和跨学科技术，包括最先进的冷冻技术 电子显微镜、X 射线晶体学、稳态动力学、体外运动测定和高分辨率 荧光显微镜下，我们将系统剖析NM2的激活和调控机制。到 为了实现这一目标，在目标 1 中，我们将确定 NM2 电机 ATP 酶循环中的主要结构状态来解释 酶的功能。在目标 2 中，我们将确定全非活性状态的高分辨率冷冻电镜结构。 长度 NM2 来解释其分子结构。在目标 3 中，我们将研究废除 能够对细胞中 NM2 丝的动力学形成非活性状态。总的来说，我们的研究将提供 更深入地了解NM2的结构、功能和调控。重要的是，这些知识将推进 我们对细胞中新出现的 NM2 功能的理解，从而为未来的发展奠定了基础 NM2 特异性疗法。

项目成果

Competition between myosin II and βH-spectrin regulates cytoskeletal tension.

肌球蛋白 II 和 βH 血影蛋白之间的竞争调节细胞骨架张力。

• 发表时间: 2023-06-27
• 影响因子: 7.7
• 作者: Ibar, Consuelo;Chinthalapudi, Krishna;Heissler, Sarah M;Irvine, Kenneth D
• 通讯作者: Irvine, Kenneth D

Cryo-EM structure of the autoinhibited state of myosin-2.

肌球蛋白 2 自抑制状态的冷冻电镜结构。

• 发表时间: 2021-12-24
• 影响因子: 13.6
• 作者: Heissler, Sarah M;Arora, Amandeep S;Billington, Neil;Sellers, James R;Chinthalapudi, Krishna
• 通讯作者: Chinthalapudi, Krishna

Structural insights into actin isoforms.

肌动蛋白亚型的结构见解。

• 发表时间: 2023-02-15
• 影响因子: 7.7
• 作者: Arora, Amandeep S;Huang, Hsiang;Singh, Ramanpreet;Narui, Yoshie;Suchenko, Andrejus;Hatano, Tomoyuki;Heissler, Sarah M;Balasubramanian, Mohan K;Chinthalapudi, Krishna
• 通讯作者: Chinthalapudi, Krishna