MRI: Acquisition of Cryo-EM for Southwest Regional Center

MRI: Acquisition of Cryo-EM for Southwest Regional Center MRI: Acquisition of Cryo-EM for Southwest Regional Center We propose the acquisition of a transmission cryo-electron microscope (cryo-EM) to be integrated into the Southwestern Center for Aberration-corrected Electron Microscopy (EM) at Arizona State University (ASU), to serve the structural biology community of the Southwestern US, a region poorly served by cryo-EM facilities. ASU has developed competitive programs in three major areas for structural biology: NMR, standard x-ray crystallography, and serial femtosecond crystallography with x-ray free electron lasers, in which we enjoy a position of world leadership, for elucidating protein structure and dynamics. The acquisition of cryo-EM to complement these existing strengths will enable new discoveries in structural biology at the cutting edge of science. Supporting technical staff and a new purpose-built high stability, electrically shielded building are already in place, and a new tenure-track faculty position dedicated to cryo-EM has been authorized for immediate hire. Many of the research projects described in this application result from the major commitment to life sciences research by ASU's President Crow, with his establishment of the Biodesign Institute in 2002, now supporting about 600 biology researchers on the ASU campus. Included among these are Prof. P. Fromme, director of the new Biodesign Center for Applied Structural Discovery (CASD); and Prof. J. Spence, faculty member in CASD, Scientific Director of the NSF-funded BioXFEL STC and PI on this application. The new cryo EM faculty member will also be appointed in CASD. ASU has a long history of leadership, innovation and discovery in the field of EM, including instrumentation development, major investments in state-of-the-art facilities, support of technical staff, faculty research programs and teaching courses, TEM Winter Schools, and our recent dramatic growth and commitment to biology. As one of the largest universities in the US, ASU is a natural location for an efficient, productive and sustainable Southwestern regional center for cryo-EM. Cryo-EM has been revolutionized recently by the development of new electron-counting area detectors and phase plates. The method provides near-atomic resolution imaging of the crucial larger biomolecules, such as membrane proteins or multi-protein complexes, which resist crystallization, and is able to provide vital tomographic imaging showing macromolecular organization within cells and viruses at high resolution in three dimensions. Quenching and trapping experiments can reveal protein conformational dynamics. Thirty projects from PIs in the Southwestern region are described which will immediately benefit from this facility, and 40 Letters of Collaboration are included. Projects span the range from amyloid fibers, membrane proteins, molecular motors, DNA nanostructures, viruses, to proteins involved in CO2 fixation and photosynthesis. Research programs into new areas, such as electron diffraction from protein nanocrystals, whole-cell imaging by STEM, further development of phase plates, and 2D crystals are also described. Cryo EM will elevate the quality and increase the output of structural biology science at ASU and across the Southwest region. Our facility will make these unique capabilities, which directly reveal the molecular mechanisms on which life is based, available to the large number of gifted researchers in the Southwestern US. The lecture courses and practical instruction currently offered in biological electron microscopy at ASU will be extended to include cryo-EM, while our internationally recognized hands-on Winter School in high-resolution TEM, offered annually since 1984 and heavily oversubscribed, will be expanded to include the most popular modes of cryo-EM analysis, single-particle imaging and tomography. The next generation of scientists and educators, including groups under-represented in STEM fields, will be actively conscripted and benefit significantly from the new research projects and funding made possible by acquisition of this instrument. Keywords: cryo-EM, structural biology, protein structure, protein dynamics, drug discovery, molecular design, structure-based design, structural biology, photosynthesis.