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

• 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
• 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

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.

项目摘要 阐明大分子或分子复合物的结构组织是分子生物学的基础之一。 逐步机械地理解其功能和活动调节。原子级的细节 通过传统的结构生物学技术，如晶体学，核磁共振， 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。为此 目的，我们采取的方法，使用超分辨率荧光显微镜映射的空间 它们各自的坐标。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性蛋白质，以及用于 超分辨率图像我们将把这种方法应用于基体-纤毛过渡区的研究， 在我们的初步研究中已经显示，它是一个调控纤毛中蛋白质运输的门结构。