Structure and function of nonmuscle myosins

非肌肉肌球蛋白的结构和功能

• 批准号: 10278968
• 负责人: KRISHNA CHINTHALAPUDI
• 金额: $ 42.54万
• 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/10278968
• 关键词: ATP phosphohydrolase; Actins; Address; Adopted; Architecture; Binding; Biochemical; Biological; Biological Assay; Biology; Blood; 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; 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; Neurologic; Nucleotides; Physiological Processes; Play; Process; Property; Protein Engineering; Proteins; Regulation; Research; Resolution; Role; Structure; Techniques; Testing; Textbooks; Therapeutic; Tissues; Work; X-Ray Crystallography; base; cell motility; computerized data processing; deafness; design; enzyme activity; flexibility; high resolution imaging; human disease; innovation; mechanotransduction; 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.

项目摘要 真核细胞的一个标志是它们能够迁移、分裂、粘附和对环境信号作出反应。 非肌肉肌球蛋白-2（NM 2）马达在这些基本细胞的许多方面发挥重要作用。 通过形成短的双极丝与肌动蛋白丝相互作用的过程。NM 2电机是二进制开关 根据细胞环境在非活动状态和活动状态之间变化。NM 2电机的精确控制 活性对于其作为肌动蛋白细胞骨架的主要调节剂的细胞功能是关键的。异常调节， NM 2旁系同源物中的突变导致包括血液和神经障碍在内的一系列疾病， 心脏病、耳聋、肾炎和癌症。因此，需要NM 2特异性疗法，但缺乏基本的治疗方法。 关于NM 2旁系同源物的结构和调节的知识是其发展的瓶颈。我们的目标是 开发详细的结构和力学的理解如何通过NM 2电机驱动力的产生 各种细胞功能。利用创新和跨学科的技术，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，体外运动试验和高分辨率 荧光显微镜，我们将系统地剖析激活和调节的机制NM 2。到 为了实现这一目标，在目标1中，我们将确定NM 2马达ATP酶循环中的主要结构状态，以解释 酶功能在目标2中，我们将确定一个高分辨率的低温EM结构的非活性状态的全， 长度NM 2来解释其分子结构。在目标3中，我们将研究废除 在细胞中形成对NM 2细丝的动力学无活性状态的能力。总的来说，我们的研究将提供一个 更深入地了解NM 2的结构、功能和调节。重要的是，这些知识将促进 我们对细胞中出现的NM 2功能的理解，从而为未来的发展奠定了基础。 NM 2特异性治疗剂。