RII Track-4: Molecular Visualization by Single Particle Cryo-Electron Microscopy

点击翻译按钮获取中文摘要

Visualization of biomacromolecules at atomic resolutions provides important informationfor the in-depth understanding of the fundamental biological processes and the rules of life.Traditionally, X-ray crystallography and nuclear magnetic resonance have been the only twoavailable structural tools that can experimentally determine three-dimensional (3D) structures ofbiomacromolecules at atomic levels. Recent advances in the instrumentation and methodology ofthe advanced microscopy technique called single-particle cryo-electron microscopy (cryo-EM) isa game-changer for molecular visualization. The cryo-EM technique has become the preferredtechnique for determining the 3D structure of large macromolecular complex or membraneproteins that have been dauntingly challenging to either of the two conventional structuraltechniques. This fellowship will support the PI to develop the cryo-EM expertise at the Cryo-EMCenter of the California Institute of Technology and apply the cryo-EM technique to the structuralcharacterization of biomolecules at the atomic level. Adding cryo-EM to the PI’s structural toolboxwill not only stimulate new research directions in the PI’s research group at the University ofNebraska-Lincoln, but also benefit many research groups across the state of Nebraska, which isthe only state that hosts a Big10 university but without expertise or instrumentation for cryo-EM.With the support of this fellowship, the PI and her group member will receivecomprehensive training on structural characterization of metalloproteins and assemblies by cryo-EM, from grid preparation, image collection to 3D map construction and model development. Inparticular, this project will focus on the structural investigation of two families of interconnectedmetalloproteins involved in bacterial redox stress response and transition metal homeostasis,respectively. The expertise in cryo-EM acquired from this training will propel the PI’s group tosuccessfully achieve research goals in both the short and long term. The structural informationresulted from the proposal will fill critical gaps in fundamental structural principles that governthe metalloproteins in the bacterial response to biotic and abiotic redox stresses and antibioticresistance. In the long run, adding the cryo-EM expertise to the PI’s interdisciplinary skillset willbetter position the PI to play a leading role in structure-mechanism investigations ofmetalloproteins in redox biology. From an educational aspect, this proposal will provide directtraining opportunities to graduate students and postdocs using the cryo-EM technique. In addition,the acquired knowledge of the application of cryo-EM will be integrated into the teaching andtraining activities conducted by the PI at UNL, aligning with the educational goal in the PI’s NSFCAREER award. Furthermore, by working with the colleagues at the University of Nebraskasystem, the PI’s cryo-EM expertise will enable new research explorations and catalyze newcollaboration across the campus and promote innovation research.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.