Structure mapping of molecular complexes by super-resolution microscopy

通过超分辨率显微镜绘制分子复合物的结构图

基本信息

批准号：
9902489
负责人：
Bo Huang
金额：
$ 34.28万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-05-04 至 2022-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9902489
关键词：
Address Affect Algorithmic Analysis Algorithms Antibodies Architecture Binding Biochemical Biological Models Biological Process Biology Cells Centrosome Chimeric Proteins Cilia Collaborations Complex Cryoelectron Microscopy Crystallography Data Development Diffusion Dimensions Disease Epitopes Explosion Face Fluorescence Fluorescence Microscopy Fluorescent Probes Focal Adhesions Gene Mutation Genes Genetic Heterogeneity Image Image Analysis Immunofluorescence Immunologic In Vitro Individual Label Macromolecular Complexes Maps Methods Microscope Microscopy Modernization Molecular Molecular Conformation Morphologic artifacts Morphology Mutation Nuclear Magnetic Resonance Nuclear Pore Complex Pathogenesis Positioning Attribute Process Proteins Regulation Resolution Ribosomes Role Signal Transduction Structure Structure-Activity Relationship Techniques Technology Tertiary Protein Structure Testing Transportation X-Ray Crystallography base ciliopathy computerized tools experience experimental study flexibility fluorophore image registration improved insight kinetosome macromolecular assembly macromolecule novel therapeutics organizational structure overexpression particle protein transport quantitative imaging reconstruction stoichiometry structural biology three dimensional structure trafficking

项目摘要

PROJECT SUMMARY Eluciding the structural organization of macromolecules or molecular complexes is one of the fundamental steps towards mechanistic understanding of their function and activity regulation. With atomic level details elucided by traditional structural biology techniques such as crystallography, nuclear magnetic resonance and cryo-electron microscopy, the next step is to place this structural information in to the cellular context. For this purpose, we take the approach of using super-resolution fluorescence microscopy to map the spatial coordinates of their individual components. For this approach, we plan to develop a scalable method for efficient alabeling endogenous proteins for super-resolution microscopy, as well as analysis algorithms for super-resolution images. We will apply this approach to the study of basal body - cilium transition zone, which has been shown in our preliminary study to be a gate structure that regulates protein trafficking in the cilium.

项目概要阐明大分子或分子复合物的结构组织是基本原理之一朝着机械性理解其功能和活动调节的步骤。具有原子级细节传统的结构生物学技术如晶体学、核磁共振和冷冻电子显微镜，下一步是将这些结构信息放入细胞环境中。为了这出于目的，我们采用超分辨率荧光显微镜的方法来绘制空间图它们各个组件的坐标。对于这种方法，我们计划开发一种可扩展的方法用于超分辨率显微镜的高效标记内源蛋白质，以及分析算法超分辨率图像。我们将应用这种方法来研究基体-纤毛过渡区，我们的初步研究已表明它是调节纤毛中蛋白质运输的门结构。