MEDIC - MicroED Imaging Center - Driving Biomedical Projects (DBPs)

推动生物医学项目（DBP） 总结 我们选择了驱动生物医学项目（DBPs），以涵盖生物医学领域的最前沿 研究并覆盖美国的东海岸到西海岸。生物化学中一些最基本的问题是 正在解决使用微晶电子衍射（MicroED）：配体如何调节结构， G蛋白偶联受体的功能（Lefkowitz，Stevens）;什么是构象动力学， 在最大的膜转运蛋白家族中参与底物识别和转运？（艾布拉姆森， Zheng）;参与神经传递和融合的大孔蛋白质复合物的结构生物学 （Hurley）;膜蛋白中的质子化和带电状态以及它们如何影响蛋白质功能（Rees， Yeager）;蛋白质如何被抗体识别以及蛋白质如何调节基因表达（Bjorkman， Feigon）;以及MicroED在理解和发现天然产物中的效用（货车der Donk）。我们选择 把我们的精力集中在研究天然产物，蛋白质复合物和膜蛋白，因为这些 是极具挑战性的，但对于药物发现和医学来说非常重要的目标。与其他人的互动 DBPs将由Gonen博士监督，他有20多年的膜工作经验 不同种类的蛋白质，包括，通道，转运蛋白和受体，并确定了几个关键 膜蛋白结构的cryoEM和X射线晶体学。此外，他还拥有化学学位， 这使他能够有效地与化学家和生物学家进行交流。许多DBP目标是 难以表达、纯化和结晶。通过应用MicroED，我们可以帮助缓解这些困难;当 如果目的膜蛋白以低量表达，则所得材料仍可能足以用于 通过MicroED进行结构测定。最后，MicroED可以从飞秒级提供原子级分辨率的结构， 大量的小分子和天然产物而无需额外的纯化和结晶。所有DBP 受益于该应用程序的技术和研究开发项目，并且是良好的测试平台， 他们它们也是传播MicroED技术的极好工具。

Driving Biomedical Projects (DBPs) Summary We selected the Driving Biomedical Projects (DBPs) for this proposal to encompass the forefront in biomedical research and cover the US from coast to coast. Some of the most fundamental questions in biochemistry are being addressed using microcrystal electron diffraction (MicroED): How do ligands regulate the structure and function of G-protein coupled receptors? (Lefkowitz, Stevens); What are the conformational dynamics that are involved in substrate recognition and transport in the largest family of membrane transporters? (Abramson, Zheng); Structural biology of large porous protein complexes that are involved in neurotransmission and fusion (Hurley); The protonation and charged state in membrane proteins and how those affect protein function (Rees, Yeager); How proteins are recognized by antibodies and how proteins can regulate gene expression (Bjorkman, Feigon); and the utility of MicroED in understanding and discovery of natural products (van der Donk). We chose to focus our efforts on the study of natural products, protein complexes and of membrane proteins because these are extremely challenging and yet very important targets for drug discovery and medicine. The interactions with the DBPs will be supervised by Dr. Gonen, who has more than 20 years' experience working with membrane proteins of different kinds, including, channels, transporters and receptors and has determined several key membrane protein structures by cryoEM and X-ray crystallography. Moreover, he holds a degree in Chemistry which allows him to effectively communicate with chemists and biologists alike. Many of the DBP targets are hard to express, purify and crystallize. By applying MicroED we help to alleviate these difficulties; when a membrane protein of interest if expressed in low amounts, the resulting material may still be sufficient for structure determination by MicroED. Finally, MicroED can deliver atomic resolution structures from femtogram amounts of small molecules and natural products without additional purification and crystallization . All DBPs benefit from the Technology and Research Development projects of this application and are good testbeds for them. They are also excellent vehicles for dissemination of MicroED technology.