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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.
Authors:
 [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
Type:
Accepted Manuscript
Journal Name:
IUCrJ
Additional Journal Information:
Journal Volume: 2; Journal Issue: 4; Journal ID: ISSN 2052-2525
Publisher:
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
Language:
English
Subject:
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:
1208832

Conrad, Chelsie E., Basu, Shibom, James, Daniel, Wang, Dingjie, Schaffer, Alexander, Roy-Chowdhury, Shatabdi, Zatsepin, Nadia A., Aquila, Andrew, Coe, Jesse, Gati, Cornelius, Hunter, Mark S., Koglin, Jason E., Kupitz, Christopher, Nelson, Garrett, Subramanian, Ganesh, White, Thomas A., Zhao, Yun, Zook, James, Boutet, Sébastien, Cherezov, Vadim, Spence, John C. H., Fromme, Raimund, Weierstall, Uwe, and Fromme, Petra. A novel inert crystal delivery medium for serial femtosecond crystallography. United States: N. p., Web. doi:10.1107/S2052252515009811.
Conrad, Chelsie E., Basu, Shibom, James, Daniel, Wang, Dingjie, Schaffer, Alexander, Roy-Chowdhury, Shatabdi, Zatsepin, Nadia A., Aquila, Andrew, Coe, Jesse, Gati, Cornelius, Hunter, Mark S., Koglin, Jason E., Kupitz, Christopher, Nelson, Garrett, Subramanian, Ganesh, White, Thomas A., Zhao, Yun, Zook, James, Boutet, Sébastien, Cherezov, Vadim, Spence, John C. H., Fromme, Raimund, Weierstall, Uwe, & Fromme, Petra. A novel inert crystal delivery medium for serial femtosecond crystallography. United States. doi:10.1107/S2052252515009811.
Conrad, Chelsie E., Basu, Shibom, James, Daniel, Wang, Dingjie, Schaffer, Alexander, Roy-Chowdhury, Shatabdi, Zatsepin, Nadia A., Aquila, Andrew, Coe, Jesse, Gati, Cornelius, Hunter, Mark S., Koglin, Jason E., Kupitz, Christopher, Nelson, Garrett, Subramanian, Ganesh, White, Thomas A., Zhao, Yun, Zook, James, Boutet, Sébastien, Cherezov, Vadim, Spence, John C. H., Fromme, Raimund, Weierstall, Uwe, and Fromme, Petra. 2015. "A novel inert crystal delivery medium for serial femtosecond crystallography". United States. doi:10.1107/S2052252515009811. https://www.osti.gov/servlets/purl/1208832.
@article{osti_1208832,
title = {A novel inert crystal delivery medium for serial femtosecond crystallography},
author = {Conrad, Chelsie E. and Basu, Shibom and James, Daniel and Wang, Dingjie and Schaffer, Alexander and Roy-Chowdhury, Shatabdi and Zatsepin, Nadia A. and Aquila, Andrew and Coe, Jesse and Gati, Cornelius and Hunter, Mark S. and Koglin, Jason E. and Kupitz, Christopher and Nelson, Garrett and Subramanian, Ganesh and White, Thomas A. and Zhao, Yun and Zook, James and Boutet, Sébastien and Cherezov, Vadim and Spence, John C. H. and Fromme, Raimund and Weierstall, Uwe and Fromme, Petra},
abstractNote = {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.},
doi = {10.1107/S2052252515009811},
journal = {IUCrJ},
number = 4,
volume = 2,
place = {United States},
year = {2015},
month = {6}
}