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Title: Atomic structure of granulin determined from native nanocrystalline granulovirus using an X-ray free-electron laser

To understand how molecules function in biological systems, new methods are required to obtain atomic resolution structures from biological material under physiological conditions. Intense femtosecond-duration pulses from X-ray free-electron lasers (XFELs) can outrun most damage processes, vastly increasing the tolerable dose before the specimen is destroyed. This in turn allows structure determination from crystals much smaller and more radiation sensitive than previously considered possible, allowing data collection from room temperature structures and avoiding structural changes due to cooling. Regardless, high-resolution structures obtained from XFEL data mostly use crystals far larger than 1 μm3 in volume, whereas the X-ray beam is often attenuated to protect the detector from damage caused by intense Bragg spots. Here, we describe the 2 Å resolution structure of native nanocrystalline granulovirus occlusion bodies (OBs) that are less than 0.016 μm3 in volume using the full power of the Linac Coherent Light Source (LCLS) and a dose up to 1.3 GGy per crystal. The crystalline shell of granulovirus OBs consists, on average, of about 9,000 unit cells, representing the smallest protein crystals to yield a high-resolution structure by X-ray crystallography to date. The XFEL structure shows little to no evidence of radiation damage and is more completemore » than a model determined using synchrotron data from recombinantly produced, much larger, cryocooled granulovirus granulin microcrystals. Furthermore, our measurements suggest that it should be possible, under ideal experimental conditions, to obtain data from protein crystals with only 100 unit cells in volume using currently available XFELs and suggest that single-molecule imaging of individual biomolecules could almost be within reach.« less
Authors:
 [1] ;  [2] ;  [2] ; ORCiD logo [3] ;  [2] ;  [4] ;  [4] ;  [5] ;  [6] ;  [5] ;  [2] ;  [7] ;  [8] ;  [9] ;  [10] ;  [11] ;  [2] ;  [11] ;  [11] ;  [12] more »;  [13] ;  [14] ;  [15] ;  [11] ;  [13] ;  [16] ;  [11] ;  [2] ;  [17] ;  [11] ;  [11] ;  [11] ;  [18] ;  [19] ;  [4] ;  [2] ; ORCiD logo [20] « less
  1. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); MRC Lab. of Molecular Biology, Cambridge (United Kingdom)
  2. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  3. The Univ. of Auckland, Auckland (New Zealand); Friedrich Miescher Institute for Biomedical Research, Basel (Switzerland)
  4. The Univ. of Auckland, Auckland (New Zealand)
  5. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); European XFEL GmbH, Hamburg (Germany)
  6. Arizona State Univ., Tempe, AZ (United States); Paul Scherrer Inst. (PSI), Villigen (Switzerland)
  7. Max Planck Institute for Medical Research, Heidelberg (Germany); Univ. of Hamburg, Hamburg (Germany)
  8. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  9. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); SLAC National Accelerator Lab., Menlo Park, CA (United States)
  10. Arizona State Univ., Tempe, AZ (United States); Max Planck Institute for Medical Research, Heidelberg (Germany)
  11. Arizona State Univ., Tempe, AZ (United States)
  12. Arizona State Univ., Tempe, AZ (United States); Univ. of Wisconsin-Milwaukee, Milwaukee, WI (United States)
  13. Max Planck Institute for Medical Research, Heidelberg (Germany)
  14. SLAC National Accelerator Lab., Menlo Park, CA (United States); National Science Foundation BioXFEL Science and Technology Center, Buffalo, NY (United States)
  15. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Paul Scherrer Inst. (PSI), Villigen (Switzerland)
  16. Arizona State Univ., Tempe, AZ (United States); ShanghaiTech Univ., Shanghai (China)
  17. SLAC National Accelerator Lab., Menlo Park, CA (United States); Brookhaven National Lab. (BNL), Upton, NY (United States)
  18. Univ. Basel, Basel (Switzerland)
  19. Julius Kuehn Institute (JKI), Darmstadt (Germany)
  20. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany); Univ. of Hamburg, Hamburg (Germany)
Publication Date:
Grant/Contract Number:
617095583; UOA1221; AC02-76SF00515
Type:
Accepted Manuscript
Journal Name:
Proceedings of the National Academy of Sciences of the United States of America
Additional Journal Information:
Journal Volume: 114; Journal Issue: 9; Journal ID: ISSN 0027-8424
Publisher:
National Academy of Sciences, Washington, DC (United States)
Research Org:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org:
USDOE
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; 59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; XFEL; nanocrystals; structural biology; bioimaging; SFX
OSTI Identifier:
1353189