Structure mapping of molecular complexes by super-resolution microscopy

项目总结 确定大分子或分子络合物的结构组织是基本的方法之一 逐步机械化地理解它们的功能和活动调节。原子级别的详细信息 由传统的结构生物学技术阐明，如结晶学、核磁共振和 冷冻电子显微镜，下一步是将这种结构信息放入细胞环境中。为了这个 目的：我们采用超分辨荧光显微技术来绘制空间图谱。 它们各自组件的坐标。对于这种方法，我们计划开发一种可伸缩的方法 用于超分辨率显微镜的高效白蛋白标记，以及分析算法 超分辨率图像。我们将把这种方法应用于基底体-纤毛过渡带的研究，该过渡带 我们的初步研究表明，它是一种门结构，调节纤毛中的蛋白质运输。

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.