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

推动生物医学项目(DBP) 摘要 我们为这项计划选择了推动生物医学项目(DBP)，以涵盖生物医学领域的前沿 从东海岸到西海岸对美国进行研究和报道。生物化学中的一些最基本的问题是 利用微晶电子衍射(MicroED)解决：配体如何调节结构和 G蛋白偶联受体的功能？(莱夫科维茨，史蒂文斯)；构象动力学是什么？ 在最大的膜转运蛋白家族中参与底物识别和运输？(艾布拉姆森， 参与神经传递和融合的大孔蛋白复合体的结构生物学 (Hurley)；膜蛋白中的质子化和带电状态及其如何影响蛋白质功能(Rees， 耶格尔)；蛋白质如何被抗体识别以及蛋白质如何调节基因表达(比约克曼， 以及微电子能谱在理解和发现天然产物方面的效用(范德顿克)。我们选择了 专注于天然产物、蛋白质复合体和膜蛋白的研究，因为这些 对于药物发现和医学来说都是极具挑战性的，但也是非常重要的目标。与以下项目的互动 DBP将由戈南博士监督，他有20多年的膜工作经验 不同种类的蛋白质，包括通道、转运体和受体，并决定了几个关键的 用低温电子显微镜和X射线结晶学研究膜蛋白结构。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。DBP的许多目标都是 难以表达、提纯和结晶。通过应用MicroED，我们帮助缓解了这些困难；当 如果以低量表达感兴趣的膜蛋白，所得到的材料可能仍然足以 用微电子能谱进行结构测定。最后，MicroED可以从Femtogram中提供原子分辨率结构 大量的小分子和天然产物，而不需要额外的提纯和结晶。所有DBP 受益于该应用程序的技术和研究开发项目，是 他们。它们也是传播微ED技术的极佳工具。

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.