Transition To Excellence: From Single-Molecule Force Spectroscopy to Single-Particle Cryogenic Electron Microscopy

这个过渡到卓越项目将支持主要研究者的专业发展，学习尖端的高分辨率显微镜技术：单粒子低温电子显微镜（SP Cryo-EM）。在该项目的学习阶段完成后，这些新技术将用于深入了解负责获得结构和大型复杂蛋白质结构维持的基本生物过程。这些过程目前对大型蛋白质的了解很少，但它们对蛋白质的正确生物功能至关重要;功能障碍可能导致许多严重疾病，包括神经退行性疾病，如帕金森病。SP Cryo-EM最近彻底改变了结构生物学，因此该项目有可能在迄今为止无法达到的三维细节水平上为蛋白质结构获取和恢复过程的基础知识做出贡献，这对于理解蛋白质折叠，错误折叠和结构恢复机制非常有用。该项目涉及SP Cryo-EM的密集理论和实验培训，并将专业推进一名高级研究员和一些博士后，博士和本科生。该项目将促进使用低温电子显微镜的新型高分辨率结构方法来表征蛋白质折叠途径，这是使用传统方法难以实现的。该项目将使首席研究员能够通过伴侣蛋白对蛋白质折叠和蛋白质结构恢复机制进行高分辨率（近原子）低温电子显微结构研究领域的研究能力进行重大扩展。这一转变将使人们对大分子内部和之间复杂的原子间相互作用有新的认识，这些相互作用决定了结构的形成和动力学，这对于获得或恢复生物功能至关重要。结合分子水平的分辨率技术，如原子力显微镜与低温电子显微镜的高分辨率结构方法，将允许详细表征蛋白质生物合成和非常大的蛋白质，如锚蛋白和萤火虫荧光素酶折叠途径之间的关系，这是以前在接近自然条件下无法以原子分辨率进行研究的。该奖项反映了NSF的法定使命，通过使用基金会的知识价值和更广泛的影响审查标准进行评估，

This Transition to Excellence Project will support the Principal Investigator’s Professional Development to learn the cutting edge high-resolution microscopy technique: Single-Particle Cryogenic Electron Microscopy (SP Cryo-EM). After the learning phase of the Project is completed, these new techniques will be applied to gain insights into fundamental biological processes responsible for the acquisition of structure and for structural maintenance of large and complex proteins. These processes are currently poorly understood for large proteins, yet they are critical for proteins’ correct biological functions; malfunction may lead to a number of serious illnesses including neurodegenerative disorders such as Parkinson’s disease. SP Cryo-EM has recently revolutionized structural biology and therefore this project has a potential to contribute to the basic knowledge of protein structure acquisition and restoration processes at a hitherto unattainable level of three-dimensional detail, which will be extremely useful for understanding protein folding, misfolding and structural recovery mechanisms. The project involves intense theoretical and experimental training in SP Cryo-EM and will professionally advance one senior investigator and a number of postdoctoral, doctoral and undergraduate students. This project will promote the use of novel high resolution structural methods of cryogenic electron microscopy to characterize protein folding pathways, which have been difficult using traditional methods. This project will allow the Principal Investigator to undergo a major expansion of research capabilities into the field of high resolution (near atomic) cryogenic electron microcopy structural studies of protein folding and protein structure recovery mechanisms by chaperones. This transition will allow new insights into complex interatomic interactions inside and between macromolecules that determine structure formation and dynamics, which are critical for the acquisition or restoration of biological function. A combination of molecular-level resolution techniques such as atomic force microscopy with high resolution structural methods of cryogenic electron microscopy will allow detailed characterization of the relationships between protein biosynthesis and folding pathways for very large proteins such as ankyrins and firefly luciferase, which could not be studied before at atomic resolution under near native conditions.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.