Intracellular transport and organelle biology at the nanoscale: A multidimensional super-resolution approach

纳米尺度的细胞内运输和细胞器生物学：多维超分辨率方法

基本信息

批准号：
10623590
负责人：
Ke Xu
金额：
$ 44.14万
依托单位：
UNIVERSITY OF CALIFORNIA BERKELEY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-15 至 2028-07-31
项目状态：
未结题

项目摘要

Project Summary My research takes a unique approach in which the development of next-generation microscopy methods progresses in parallel with fundamental discoveries in cell biology. On the method-development front, besides earlier success in achieving sub-10 nm resolution for super-resolution microscopy, my recent work has pioneered the concept of multidimensional and multifunctional super-resolution microscopy, in which intracellular functional parameters, including local chemical polarity, pH, diffusivity, and protein activity, are mapped out at nanometer resolution and single-molecule sensitivity. Empowered by such capabilities, my lab has been highly successful in unveiling hidden subcellular structures and processes, as well as their underlying biophysical rules, for diverse systems ranging from the mammalian cytoskeleton, intracellular transport, organelle morphology and biogenesis, to membrane biology. Our future research continues to push forward the synergy between method development and biological discoveries. Major directions include charge-modulated protein interactions and effects on diffusion inside the organelle lumen, superdiffusion and subdiffusion in the living cell, organelle pH dynamics and role in protein trafficking, and the structure and physical properties of the ER exit site in relationship with the biogenesis of transport carriers. Moreover, by integrating super-resolution microscopy with FIB-SEM, we will obtain holistic pictures of the unusually thin tubular organelles we recently discovered and further substantiate their functions and biogenesis. Separately, we are developing a new tool, single-molecule electrophoresis microscopy, to quantify protein charge states at the super-resolution level. Together, through the continued development of empowering microscopy tools and their tactical application to fundamental biological questions, we will continue shifting the paradigms of how we understand the complex, dynamic behavior of the cell.

项目概要我的研究采用了一种独特的方法，其中下一代显微镜方法的开发与细胞生物学的基本发现并行进展。在方法开发方面，除了我早期的工作是在超分辨率显微镜中实现亚 10 nm 分辨率，我最近的工作是开创了多维多功能超分辨率显微镜的概念，其中细胞内功能参数，包括局部化学极性、pH、扩散率和蛋白质活性以纳米分辨率和单分子灵敏度绘制。有了这样的能力，我的实验室在揭示隐藏的亚细胞结构和过程及其基础方面取得了巨大成功生物物理规则，适用于从哺乳动物细胞骨架、细胞内运输、细胞器形态和生物发生，到膜生物学。我们未来的研究将继续推动方法开发和生物学发现之间的协同作用。主要方向包括电荷调制蛋白质相互作用以及对细胞器腔内扩散、细胞器内超扩散和亚扩散的影响活细胞、细胞器 pH 动力学和蛋白质运输中的作用，以及细胞器的结构和物理特性 ER 出口位点与转运载体的生物发生有关。此外，通过集成超分辨率通过 FIB-SEM 显微镜，我们将获得最近我们研究的异常薄的管状细胞器的整体图像发现并进一步证实了它们的功能和生物发生。另外，我们正在开发一个新工具，单分子电泳显微镜，以超分辨率水平量化蛋白质电荷状态。共同努力，通过不断开发增强显微镜工具及其战术应用，基本的生物学问题，我们将继续改变我们理解复杂的范式，细胞的动态行为。