Spatial Mutation of Membrane Protein Assembly Dynamics Using Nano-Actuators

描述（由申请人提供）：血管生成素通过配体诱导的内皮细胞组装和Tie2受体易位刺激内皮细胞的促血管生成信号，并调节内皮细胞的增殖、迁移和组装等几个关键过程。然而，目前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，如维管发芽、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分子机制，必须对Tie2的时空动态进行高分辨率实时监测。本文提出了一种创新的方法，利用新型磁等离子体纳米驱动系统来研究Tie2的动态组装和空间分布变化。与传统显微镜只能提供细胞状态的静态评估不同，该系统允许我们在整个血管生成期间以及整个形态发生发展过程中连续可视化信号分子的生物分子事件。此外，利用纳米驱动系统的磁镊能力，对Tie2的空间分布进行了控制

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?