Near-atomic cryo-EM imaging of a small protein displayed on a designed scaffolding system

Journal Article · Mon Mar 05 00:00:00 EST 2018 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1718825115· OSTI ID:1423732

Liu, Yuxi ^[1]; Gonen, Shane ^[2]; Gonen, Tamir ^[3]; Yeates, Todd O. ^[4]

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1. Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095,
2. Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, VA 20147,, Howard Hughes Medical Institute, University of California, San Francisco, CA 94143,
3. Howard Hughes Medical Institute, Department of Physiology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095,, Howard Hughes Medical Institute, Department of Biological Chemistry, David Geffen School of Medicine, University of California, Los Angeles, CA 90095,
4. Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095,, UCLA-DOE Institute for Genomics and Proteomics, Los Angeles, CA 90095,, UCLA Molecular Biology Institute, Los Angeles, CA 90095

Significance New electron microscopy (EM) methods are making it possible to view the structures of large proteins and nucleic acid complexes at atomic detail, but the methods are difficult to apply to molecules smaller than approximately 50 kDa, which is larger than the size of the average protein in the cell. The present work demonstrates that a protein much smaller than that limit can be successfully visualized when it is attached to a large protein scaffold designed to hold 12 copies of the attached protein in symmetric and rigidly defined orientations. The small protein chosen for attachment and visualization can be modified to bind to other diverse proteins, opening a new avenue for imaging cellular proteins by cryo-EM.

• Sponsoring Organization: USDOE Office of Science (SC), Biological and Environmental Research (BER)
• Grant/Contract Number: FC02-2ER63421
• OSTI ID: 1423732
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Vol. 115 Journal Issue: 13; ISSN 0027-8424
• Publisher: Proceedings of the National Academy of Sciences
• Country of Publication: United States
• Language: English