Cytoskeletal Motors and Scaffolds in Membrane Dynamics and Motility

膜动力学和运动中的细胞骨架马达和支架

• 批准号: 9143338
• 负责人: E. Michael Ostap
• 金额: $ 7.24万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-26 至 2019-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9143338
• 关键词: Actins; Adhesions; Autophagocytosis; Binding; Biochemical; Biochemistry; Biological; Cardiovascular Diseases; Cell physiology; Cells; Cellular biology; Collection; Complex; Cytoskeletal Proteins; Cytoskeleton; Defect; Dependence; Development; Dissociation; Dynein ATPase; Early Endosome; Endocytosis; Endosomes; Event; Family; Filament; Fluorescence; Fluorescence Polarization; Geometry; Goals; Head; Healthcare; Hearing; Image; In Vitro; Intracellular Membranes; Kinesin; Kinetics; Life; Link; Malignant Neoplasms; Measures; Mechanics; Membrane; Membrane Proteins; Microfabrication; Microfilaments; Microscopy; Microtubules; Molecular; Molecular Motors; Morphogenesis; Motion; Motor; Movement; Myosin ATPase; Myosin Type I; Neurons; Organelles; Pathologic Processes; Pathology; Property; Protein Isoforms; Proteins; Recruitment Activity; Regulation; Research; Research Personnel; Resolution; Role; Scaffolding Protein; Sorting - Cell Movement; Stress; Techniques; Technology; Tertiary Protein Structure; Testing; Tissues; Total Internal Reflection Fluorescent; Transmembrane Transport; Work; Working stroke; Wound Healing; X-Ray Crystallography; arm; base; biophysical properties; cell motility; member; membrane assembly; motor control; nanometer; nexin; optical traps; polymerization; protein function; public health relevance; reconstitution; scaffold; single molecule; structural biology; technology development; trafficking

DESCRIPTION (provided by applicant): The microtubule and actin cytoskeletons are essential for the movement, polarization, sorting, and morphogenesis of intracellular membrane compartments. Molecular motors, scaffolding proteins, and associated filaments are recruited to membranes to power diverse trafficking events that have different force, power, kinetic, and transport requirements. Defining the mechanisms of trafficking requires understanding how different motor isoforms and families work alone and in teams, and how motors and filaments work with the scaffolding proteins and adapters that link them to membranes. Determining these mechanisms requires a detailed understanding of: the cellular organization and dynamics of the cytoskeleton and membranes; the spatial, kinetic, and mechanical relationship of different motors and filaments; the structural and biophysical properties of cytoskeletal-membrane assemblies; and the biophysical parameters that define the capabilities and mechanisms of motors and scaffolds when operating under working conditions. To this end, we assembled an extraordinarily strong scientific team with expertise in cell biology, biochemistry, structural biology, structural dynamics, and technology-development to define the role of the cytoskeleton and molecular motors in trafficking. Our team includes pioneers in the use of state-of-the-art imaging, single-molecule, and structural techniques to discover how cytoskeletal proteins function in complex cellular events. High-resolution live-cell microscopy, reconstituted cytoskeletal geometries using microfabrication and dielectrophoresis, X-ray crystallography, nanometer-resolved fluorescence tracking, single-molecule fluorescence polarization, optical trapping, and advanced biochemical techniques will be applied in highly collaborative studies to understand how motors, scaffolds, and filaments work together to power membrane dynamics. The projects and investigators are interdependent and are closely linked through research goals and common technologies, and the Aims were formulated to capitalize on the unique strengths of the team members while taking advantage of extensive synergies between the groups. We will focus on the following four Aims: (1) Investigate the Dynamics of Molecular Motors in Organelle Transport and Membrane Remodeling; (2) Investigate the Structural, Biochemical, and Cellular Properties of Cytoskeleton-Membrane Scaffolds in Organelle Morphogenesis and Motility; (3) Discover the Mechanical and Biochemical Adaptations of Membrane- Associated Motors and Scaffolds; (4) Investigate the Structural Dynamics of Myosin, Dynein and Motor Collections.

描述（由申请人提供）：微管和肌动蛋白细胞骨架对于细胞内膜区室的运动、极化、分选和形态发生至关重要。分子马达、支架蛋白和相关的细丝被招募到膜上，为具有不同力、功率、动力学和运输要求的各种运输事件提供动力。定义贩运机制需要了解不同的运动亚型和家族如何单独和团队工作，以及运动和细丝如何与将它们连接到膜的支架蛋白和适配器一起工作。确定这些机制需要详细了解：细胞骨架和膜的细胞组织和动力学;不同马达和细丝的空间，动力学和机械关系;细胞膜组件的结构和生物物理特性;以及定义马达和支架在工作条件下操作时的能力和机制的生物物理参数。为此，我们组建了一支非常强大的科学团队，他们拥有细胞生物学，生物化学，结构生物学，结构动力学和技术开发方面的专业知识，以确定细胞骨架和分子马达在贩运中的作用。我们的团队包括使用最先进的成像，单分子和结构技术来发现细胞骨架蛋白在复杂细胞事件中的功能的先驱。高分辨率活细胞显微镜，使用微加工和介电泳，X射线晶体学，纳米分辨荧光跟踪，单分子荧光偏振，光学捕获和先进的生物化学技术重建细胞骨架几何形状将应用于高度合作的研究，以了解电机，支架和细丝如何共同工作，为膜动力学提供动力。项目和研究人员是相互依赖的，并通过研究目标和共同技术紧密联系在一起，制定目标是为了利用团队成员的独特优势，同时利用团队之间的广泛协同作用。本课程将围绕以下四个方面展开：（1）研究细胞器运输和膜重构中的分子马达动力学;（2）研究细胞器-膜支架在细胞器形态发生和运动中的结构、生物化学和细胞特性;（3）发现膜相关马达和支架的机械和生物化学适应;（4）研究细胞器-膜支架的结构和生物化学适应。（4）研究肌球蛋白、动力蛋白和运动集合的结构动力学。