Spatial Mutation of Membrane Protein Assembly Dynamics Using Nano-Actuators

使用纳米致动器的膜蛋白组装动力学的空间突变

• 批准号: 8918731
• 负责人: Young-wook Jun
• 金额: $ 22.5万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8918731
• 关键词: Address; Angiopoietins; Behavior; Biological; Biology; Blinking; Blood Vessels; Cells; Characteristics; Complex; Coupling; Development; Diagnostic; Disease; EGF-Like Domain; Endothelial Cells; Environment; Event; Goals; Health; Heterogeneity; Image; Imagery; Immunoglobulins; Individual; Individuality; Life; Ligand Binding; Ligands; Link; Location; Magnetism; Malignant Neoplasms; Maps; Membrane; Membrane Proteins; Microscopy; Molecular; Monitor; Morphogenesis; Mutation; Optics; Photobleaching; Process; Property; Protein Tyrosine Kinase; Resolution; Role; Signal Transduction; Signaling Molecule; Spatial Distribution; System; TIE-2 Receptor; Techniques; Technology; Testing; Therapeutic; Time; Transducers; angiogenesis; base; cell behavior; extracellular; imaging system; innovation; insight; interest; migration; nano; nanoparticle; nanoprobe; novel; optical imaging; particle; plasmonics; receptor; research study; response; segregation; single molecule; spatiotemporal; temporal measurement; tool

DESCRIPTION (provided by applicant): Angiopoietins stimulate proangiogenic signals of endothelial cells (ECs) via ligand-induced assembly and translocation of Tie2 receptors and modulate several key processes in ECs including proliferation, migration, and assembly. However, it is still unclear how these biomolecular regulators coordinate highly complex morphogenesis such as vascular sprouting, migration, branching, and network formation. To understand molecular mechanisms of angiogenesis with high spatio-temporal complexity, the spatiotemporal dynamics of Tie2 must be monitored in real-time with high resolution. Here, I propose an innovative approach to investigate dynamic assembly and spatial distribution changes of Tie2 using a new magneto-plasmonic nano-actuation system. Unlike conventional microscopy that only provides a static assessment of cell status, the proposed system allows us to visualize biomolecular events of the signaling molecules continuously over the entire angiogenesis period as well as throughout morphogenetic development. Additionally, by using magnetic tweezing capability of the nano-actuation system, control of Tie2 spatial distributions is possible, enabling the study of how spatially polarized Tie2 distribution influences the cell's behavior and fate. With this new technique, specifically, I will challenge the following questions: 1. How do the assembly state and distribution of Tie2 change after stimulation with various ligands such as angiopoietins? 2. Are the assembly states of Tie2 related to the different subcellular locations such as cell periphery, cell-to-cell contacts, and cell-to-substratum contacts? 3. How do spatially polarized distributions of Tie2 influence a cell's behavior?

描述（由申请人提供）：血管生成素通过配体诱导的TIE2受体的组装和易位刺激内皮细胞（EC）的促血管生成信号，并调节EC中的几个关键过程，包括增殖，迁移和组装。然而，尚不清楚这些生物分子调节剂如何协调高度复杂的形态发生，例如血管发芽，迁移，分支和网络形成。为了了解具有高时空复杂性的血管生成的分子机制，必须通过高分辨率实时监测TIE2的时空动力学。在这里，我提出了一种创新的方法，可以使用新的磁质质量纳米效应系统研究TIE2的动态组装和空间分布变化。与仅提供细胞状态的静态评估的常规显微镜不同，该系统使我们能够在整个血管生成期以及整个形态发生发育中连续地可视化信号分子的生物分子事件。此外，通过使用纳米实现系统的磁性镊子能力，对TIE2空间分布的控制是 可能，可以研究空间极化的TIE2分布如何影响细胞的行为和命运。具体来说，有了这种新技术，我将挑战以下问题： 1。在用各种配体（例如血管生成素）刺激后，TIE2的组装状态和分布如何变化？ 2。TIE2的组装状态是否与不同的亚细胞位置有关，例如细胞周围，细胞间接触和细胞到囊膜的接触？ 3. TIE2的空间极化分布如何影响细胞的行为？