Micro Electron Diffraction of Toxic and/or Infectious Macromolecular Nanoassemblies

项目总结 尽管结构生物学取得了现代进展，但许多与生物医学相关的结构 大分子组装仍然遥不可及或缺乏原子分辨率细节。此外， 确定全新结构的过程仍然很费力。所需的大晶体 传统的结晶学实验是一项艰巨的任务， 用X射线结晶学分析小晶体或不完美晶体的结构仍然有限。冷藏- 电子显微镜(冷冻-EM)方法有望为高通量带来新的生命 大分子结构测定的方法。低温电磁革命带来了 有了它，包括微电子衍射(MicroED)在内的新的高分辨率方法。MicroED 利用电子和纳米级三维晶体之间的强相互作用 利用新兴的冷冻-EM仪器来对抗已有的结晶学知识。 我的团队帮助MicroED取得了里程碑式的发现，并确定了全新的发现 来自晶体和晶体碎片的蛋白质结构小于可见光波长 光，原子分辨率。这些技术进步，加上更多可用的 先进的低温电磁仪器的出现，为进一步提高高能电子显微镜的性能提供了机会。 确定吞吐量结构。发展新的、更有效的方法来 利用微电子能谱进行结构测定可能为全面探索新途径开辟新途径 复杂的大分子结构，仍然是标准方法无法达到的。这些 系统包括生长小的、脆弱的或不完美的晶体的大分子络合物。这个 与这些类型的组件相关的生物医学问题多种多样，并可能广泛地 影响生物医学，既通过对疾病的基本解释，也作为治疗 站台。具体地说，我们的目标是感染性和/或有毒的丝状纳米组件 与淀粉样变性疾病有关。通过我们的努力确定这些具有挑战性的 结构，我们找到了指导低温EM改进和发展的灵感 技术，特别是微电子能谱。怀着这个目标，我的团队将承担这一发展 新的高通量晶体结构测定方法，同时获得 以真正的原子分辨率压制生物医学目标的结构。

PROJECT SUMMARY Despite modern advances in structural biology, structures of many biomedically relevant macromolecular assemblies remain out of reach or lack atomic resolution detail. In addition, the process of determining entirely novel structures remains laborious. Large crystals required for conventional crystallography experiments are a challenge to grow, and determination of structures from small or imperfect crystals by x-ray crystallography remains limited. Cryo- electron microscopy (cryo-EM) methods promise to bring new life to high-throughput approaches in macromolecular structure determination. The cryo-EM revolution has brought with it new high-resolution methods including micro electron diffraction (MicroED). MicroED exploits the strong interaction between electrons and nano-scale three-dimensional crystals by leveraging emerging cryo-EM instrumentation against established crystallographic knowledge. My group has helped achieve milestone discoveries in MicroED and determined entirely new protein structures from crystals and crystal fragments smaller than the wavelength of visible light, at atomic resolution. These technological advances, coupled with the greater availability of advanced cryoEM instruments, present an opportunity for further improvement of high- throughput structure determination. The development new and more efficient approaches to structure determination by MicroED could open new avenues for comprehensive exploration of complex macromolecular structures that remain out of reach for standard methods. These systems include macromolecular complexes that grow small, fragile, or imperfect crystals. The biomedical problems associated with these types of assemblies are varied and could broadly impact biomedicine, both through the basic interpretation of disease and as therapeutic platforms. Specifically, we aim to target infectious and/or toxic filamentous nanoassemblies associated with amyloid disease. Through our efforts in determining these challenging structures, we find inspiration to guide the improvement and development of cryoEM techniques, particularly MicroED. With this goal in mind, my group will take on the development of new high-throughput methods for crystallographic structure determination while obtaining structures of pressing biomedical targets at truly atomic resolution.