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Title: A novel inert crystal delivery medium for serial femtosecond crystallography

Serial femtosecond crystallography (SFX) has opened a new era in crystallography by permitting nearly damage-free, room-temperature structure determination of challenging proteins such as membrane proteins. In SFX, femtosecond X-ray free-electron laser pulses produce diffraction snapshots from nanocrystals and microcrystals delivered in a liquid jet, which leads to high protein consumption. A slow-moving stream of agarose has been developed as a new crystal delivery medium for SFX. It has low background scattering, is compatible with both soluble and membrane proteins, and can deliver the protein crystals at a wide range of temperatures down to 4°C. Using this crystal-laden agarose stream, the structure of a multi-subunit complex, phycocyanin, was solved to 2.5 Å resolution using 300 µg of microcrystals embedded into the agarose medium post-crystallization. The agarose delivery method reduces protein consumption by at least 100-fold and has the potential to be used for a diverse population of proteins, including membrane protein complexes.
 [1] ;  [2] ;  [2] ;  [3] ;  [2] ;  [2] ;  [2] ;  [4] ;  [2] ;  [5] ;  [4] ;  [6] ;  [7] ;  [2] ;  [2] ;  [5] ;  [5] ;  [2] ;  [6] ;  [8] more »;  [2] ;  [2] ;  [2] ;  [2] « less
  1. Arizona State Univ., Tempe, AZ (United States);The Biodesign Inst., Tempe, AZ (United States)
  2. Arizona State Univ., Tempe, AZ (United States); The Biodesign Inst., Tempe, AZ (United States)
  3. Arizona State Univ., Tempe, AZ (United States)
  4. SLAC National Accelerator Laboratory, Menlo Park, CA (United States)
  5. Deutsches Elektronen-Synchrotron (DESY), Hamburg (Germany)
  6. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  7. Univ. of Wisconsin-Milwaukee, Milwaukee, WI (United States); The Biodesign Inst., Tempe, AZ (United States)
  8. Univ. of Southern California, Los Angeles, CA (United States)
Publication Date:
Grant/Contract Number:
AC03-76SF00515; SC0010575
Accepted Manuscript
Journal Name:
Additional Journal Information:
Journal Volume: 2; Journal Issue: 4; Journal ID: ISSN 2052-2525
International Union of Crystallography
Research Org:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
59 BASIC BIOLOGICAL SCIENCES; serial femtosecond crystallography; viscous crystal delivery; protein complexes; membrane proteins; femtosecond studies; nanocrystals; coherent X-ray diffractive imaging; free-electron laser
OSTI Identifier: