MEDIC - MicroED Imaging Center at UCLA

MEDIC - 加州大学洛杉矶分校 MicroED 成像中心

基本信息

批准号：
10155524
负责人：
Tamir Gonen
金额：
$ 122.83万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10155524
关键词：
3-Dimensional Automobile Driving Biochemistry Biological Biomedical Research Categories Collaborations Communities Country Coupled Cryoelectron Microscopy Crystallization Crystallography Development Disease Educational workshop Electron Diffraction Microscopy Electrons Engineering Goals Growth Human Resources Institution Life Light Macromolecular Complexes Methodology Methods Modality Molecular Biology Molecular Structure Natural Products Phase Preparation Procedures Resolution Resources Roentgen Rays Sampling Structure System Technology Testing Time Toxin Training Visible Radiation X ray diffraction analysis community engagement electron diffraction experience improved innovation instrument instrumentation macromolecular assembly method development nanocrystal nanoscale next generation novel novel therapeutics programs protein structure screening small molecule symposium technology development tool

项目摘要

MEDIC – MicroED Imaging Center at UCLA Project Summary We are proposing to establish a national resource at UCLA for the development, training and dissemination of advanced electron diffraction technologies. Cryo-electron microscopy (cryoEM) methods promise new life to high-throughput macromolecular structure determination. CryoEM overcomes the fundamental barrier to X-ray diffraction determination of macromolecular complexes: growing X-ray grade crystals. Fortunately, the emergence of microcrystal electron diffraction (MicroED) facilitates the determination of new protein structures at atomic resolution from vanishingly small crystals. MicroED exploits the strong interaction between electrons and nano-scale three-dimensional crystals and takes advantage of emerging cryoEM instrumentation coupled to established crystallographic methods. We pioneered MicroED to achieve milestone discoveries, namely we determined previously unknown protein structures at atomic resolution from crystals smaller than the wavelength of visible light. 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 of new and more efficient approaches to structure determination by MicroED opens 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. Our proposed innovations to technology development in MicroED are in four categories: (1) improved sample screening and preparation, (2) novel refinement and phasing methods, (2) demonstrating the applicability of MicroED to natural products and small molecules, and (4) engineering new hardware for investigating structure dynamics in real time. The biomedical problems associated with these types of assemblies are broad and impact biomedicine, both through the basic understanding of disease and through enabling new therapeutic platforms. Our group of key personnel brings together expertise from all modalities of cryoEM, crystallography, biochemistry, molecular biology, and computation to help develop the next generation of MicroED tools. This effort will be informed by projects led by expert groups around the country working on projects that pose significant structural challenges. Together, we propose a resource that can make nanocrystallography and MicroED routine methodologies for non-experts. Extensive user training and community engagement will further disseminate the MicroED technology and bring new structures to light.

MEDIC – 加州大学洛杉矶分校的 MicroED 成像中心项目概要我们提议在加州大学洛杉矶分校建立一个国家资源，用于开发、培训和传播先进的电子衍射技术为冷冻电子显微镜（cryoEM）方法带来了新的生命。高通量大分子结构测定克服了 X 射线的根本障碍。大分子复合物的衍射测定：生长的 X 射线级晶体幸运的是，微晶电子衍射（MicroED）的出现促进了新蛋白质结构的测定 MicroED 利用电子之间的强相互作用，以原子分辨率从极小的晶体中进行分析。和纳米级三维晶体，并利用新兴的冷冻电镜仪器耦合我们开创了 MicroED 来实现里程碑式的发现，即我们从小于波长的晶体中获得原子分辨率的先前未知的蛋白质结构这些技术进步，加上先进冷冻电镜的可用性更高。仪器，为进一步改进高通量结构测定提供了机会。 MicroED 开发新的、更有效的结构测定方法开辟了新的途径用于全面探索仍达不到标准的复杂大分子结构这些系统包括生长小、脆弱或不完美晶体的大分子复合物。 MicroED 技术开发的拟议创新分为四类：(1) 改进的样本筛选和制备，(2) 新颖的精制和定相方法，(2) 证明其适用性 MicroED 应用于天然产物和小分子，以及 (4) 设计用于研究结构的新硬件与这些类型的组件相关的生物医学问题是广泛且具有影响力的。生物医学，既通过对疾病的基本了解，又通过启用新的治疗平台。我们的关键人员团队汇集了冷冻电镜、晶体学、生物化学、分子生物学和计算有助于开发下一代 MicroED 工具。全国各地的专家组领导的项目将为这些工作提供信息，这些项目我们共同提出了一种可以制造纳米晶体学和针对非专家的 MicroED 常规方法将进一步得到广泛的用户培训和社区参与。传播 MicroED 技术并揭示新结构。